Contact - Institut Pasteur

Transcription

S
ince the beginning, more than a century ago, biomedical research is
the main mission of the Institut Pasteur de Lille, a private foundation
created by the Mayor of Lille in 1894 to face the devastating scourge
of diphtheria and other infectious diseases. In the footsteps of their original
founders who made considerable medical breakthroughs, such the
development of the BCG vaccine for protection against tuberculosis, all the
scientists, engineers, technicians and administrative staff devote their time and
energy to the progresses in molecular medicine. In addition to infectious diseases,
the historical research topic, cancer, cardiovascular and neurodegenerative diseases are
now part of the principle research areas of the Institut Pasteur de Lille.
During the last two years, all the research units hosted on the campus of the Institut Pasteur de Lille have undergone
a major scientific reorganization in order to further increase efficiency and international visibility. Today, the institute
hosts six research units gathering 34 research teams with specific goals, four years programs and attractive working
conditions. This organization has been examined by the French national evaluation agency (AERES 1) in 2009. The
agency confirmed the excellence of both the research and the organisation. Four units were ranked A+, and two were
ranked A, representing the best results among the scientific campuses North of Paris.
This new organisation is now structured in three main areas : (i) infection and immunity, (ii) cancer and (iii)
cardiovascular, metabolic and neurodegenerative diseases. This grouping is expected to favour improved interactions
within the different areas, to increase the critical mass of scientists, engineers and technicians, and to thereby enhance
the efficiency of the research. As shown in this scientific report, most of our research is published in high-ranking
journals, including Nature, Science and Cell. Time magazine and the French journal La Recherche have cited one of
our studies 2 as one of the top-ten global scientific discoveries in 2009.
Beside these well-recognized actions, the Institut Pasteur de Lille is also committed to foster new projects, based on
promising young investigators, including at-risk initiatives. These scientific initiatives, we hope, will open new avenues
in basic and clinical research for the coming years. Our strong interactions with the scientific environment (the scientific
national agency Aviesan 3, the University of Lille, North of France and the University Hospital) allow us to develop an
integrated and global approach from basic research to translational research and medical applications, paving the
way for personalized medicine, the next challenge of the century.
1. Agence d’Evaluation de la Recherche et de l’Enseignement Supérieur
2. Nat Genet. 2009, 41:1094-9.
3. Alliance pour les sciences de la VIE et de la SANté
Philippe Amouyel, MD, PhD
General Director of the Foundation
2
Research Report 2008/2010
Contents
Contents
Infection and Immunity
Center of Infection and Immunity of Lille (C I I L)
p9
Inserm U 1019, CNRS UMR 8204, Institut Pasteur de Lille, University Lille Nord de France
affiliated to IFR 142
Camille LOCHT
composed to 11 teams :
Biology of the Pathogen
Team 1 : Molecular and Cellular Biology of Toxoplasma gondii (Stan Tomavo)
Team 2 : Molecular Biology of Schistosome Development and Reproduction (Ray Pierce)
p 13
p 16
Strategies of Infection
Team 3 : Plague and Yersinia pestis (Florent Sebbane)
Team 4 : Bacterial Respiratory Infections : Pertussis and Tuberculosis (Camille Locht)
Team 5 : Cellular microbiology of infectious pathogens (Frank Lafont)
Team 6 : Molecular and Cellular Virology of Hepatitis C (Jean Dubuisson)
p 22
p 26
p 34
p 38
Host Response and Inflammatory Processes
Team 7 : NODS-Like Receptors in Infection and Immunity (Mathias Chamaillard)
Team 8 : Lung Infection and Innate Immunity (François Trottein)
Team 9 : Lactic Acid Bacteria and Mucosal Immunity (Bruno Pot)
Team 10 : Basic and Clinical Immunology of Parasitic Diseases (Sylviane Pied)
Team 11 : Pulmonary Immunity (Anne Tsicopoulos)
p 43
p 46
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p 59
Cancer
Genetic, Functionnal and Structure Approaches of Cancer Biology
p 67
CNRS UMR 8161, Institut Pasteur de Lille, University Lille Nord de France, affiliated to IFR 142
Yvan de LAUNOIT
Team 1 : Virus, Cancer and Transcription (Yvan de Launoit)
Team 2 : Functional Studies of Tumor Suppressor Gene HIC1 (Dominique Leprince)
Team 3 : VE-statin/egf17 and vascular development (Fabrice Soncin)
Team 4 : Cancer Biology and Chemistry (Oleg Melnyk)
Team 5 : Initiation of Epithelial Cancers (Corinne Abbadie)
Team 6 : Signalling, Apoptosis and Cancer (David Tulasne)
Laboratory of Toxicology
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p 101
EA 2690, Institut Pasteur de Lille, University Lille Nord de France
Daniel MARZIN
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Cardiovascular, Metabolic and Neurodegenerative Diseases
Public Health and Molecular Epidemiology of Aging-Related Diseases
p 107
Inserm U 744, Institut Pasteur de Lille, University Lille Nord de France affiliated to IFR 142
Philippe AMOUYEL
• Team 1 : Public Health and Epidemiology of Vascular Diseases (Philippe Amouyel)
• Team 2 : Search for the Molecular Determinants of Cardiovascular Diseases via Proteomic
Analysis and Candidate Gene Approaches (Florence Pinet)
• Team 3 : Search for the Molecular Determinants of Neurodegenerative Diseases
via Transcriptomic and Candidate Gene Approaches (Jean-Charles Lambert)
Genomics and Metabolic Diseases
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p 123
p 129
p 133
CNRS UMR 8199 , Institut Pasteur de Lille, University Lille Nord de France, affiliated to IFR 114
Philippe FROGUEL
Nuclear Receptors, Cardiovascular Diseases and Diabete
p 145
Inserm U 1011, Institut Pasteur de Lille, University Lille Nord de France, affiliated to IFR 114 et IFR 142
Bart STAELS
• Team 1 : Nuclear Receptors in the Metabolic Syndrome (Bart Staels)
• Team 2 : Molecular Control of Monocyte / Macrophage Functions in Cardiometabolic
Syndrome (Giulia Chinetti)
• Team 3 : Immuno-Inflammation and Cardiovascular diseases (David Dombrowicz)
• Team 4 : Molecular Analysis of Gene Regulation in Cardiometabolic Diseases (Philippe Lefebvre)
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Cross-Sectional and Emerging
Biostructures and Molecular Drug Discovery
p 167
Inserm U 761, Institut Pasteur de Lille, University Lille Nord de France, affiliated to IFR 114 and IFR 142
Benoît DEPREZ
Inhibition of Nonsense-Mediated mRNA Decay
p 173
Groupe Avenir Inserm, Institut Pasteur de Lille, affiliated to IFR 142
Fabrice LEJEUNE
Microbiological Safety Unit
p 175
Institut Pasteur de Lille
Michèle VIALETTE
Biology and Diversity of Emerging Eukaryotic Pathogens
p 179
EA 4547, Institut Pasteur de Lille, University Lille Nord de France, affiliated to IFR 142
Eric VISCOGLIOSI
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Technological Facilities
Plethysmography/Functionnal Investigation of Airway Diseases
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Inserm U 1011, Institut Pasteur de Lille, University Lille Nord de France
David DOMBROWICZ
Microscopy - Imaging -Cytometry of the Pasteur Lille Campus (MICPaL)
p 191
facility
Frank LAFONT
Surface Plasmon Resonance
p 195
CNRS UMR 8161, Institut Pasteur de Lille, University Lille Nord de France
Marc AUMERCIER
Transcriptomics and Applied Genomics Group (TAG)
p 197
Yves LEMOINE
Peptides Chemistry, Systems, Biology
p 201
Oleg MELNYK - Hervé DROBECQ
Macromolecular Crystallography
p 207
Vincent VILLERET
HTS and ADME-PK Screening Lab
p 209
Inserm U 761, Institut Pasteur de Lille, University Lille Nord de France
Benoît DEPREZ
Nuclear Magnetic Resonance
p 211
Guy LIPPENS
Animal Unit
p 215
Jean-Pierre DECAVEL
High Security Laboratory
p 217
Jean-Pierre DECAVEL
Genomic Analysis Laboratory
p 219
Inserm U 744, Institut Pasteur de Lille , University Lille Nord de France
Philippe AMOUYEL & Nathalie FIEVET -VERRECAS
High Throughput Genomics Platform
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Genoscreen SAS
André TORDEUX
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Infection
and Immunity
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Center of Infection
and Immunity of Lille
(C I I L)
Inserm U 1019, CNRS UMR 8204,
Institut Pasteur de Lille,
University Lille Nord de France
Camille LOCHT
Contact :
00 33 3 20 87 11 51
[email protected]
Group members common facilities :
Colson-Laleu Claudine, Administrative Staff IPL
Dewailly Véronique, Administrative Staff Inserm
Evrard Edith, Administrative Staff Inserm
Houze Maria, Administrative Staff IPL
Neyrinck Jean-Loup, Administrative Responsible IPL
Renaud Marie-Christine, Administrative Staff IPL
Vantouroux Nadine, Administrative Staff IPL
Barois Nicolas, IR Inserm
Janel Sébastien, IE CNRS
Dannel Gaétane, AGT2 IPL
Flipo Joëlle, AGT2 IPL
Pollet Myriam, AGT3 IPL
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11 Teams
Biology of the Pathogen
• Molecular and Cellular Biology of Toxoplasma gondii
Stan TOMAVO
Contact : 00 33 3 20 87 74 29 - [email protected]
• Molecular Biology of Shistosome Development and Reproduction
Raymond PIERCE
Strategies of Infection
• Plague and Yersinia pestis
Florent SEBBANE
Contact : 00 33 3 20 87 11 93 - fl[email protected]
• Bacteria Respiratory Infections : Pertussis and Tuberculosis
Camille LOCHT
• Cellular microbiology of infectious pathogens
Frank LAFONT
• Molecular and Cellular Virology of Hepatitis C
Jean DUBUISSON
Host Response and Inflammatory Processes
• NODS-Like Receptors in Infection and Immunity
Mathias CHAMAILLARD
Contact : 00 33 3 20 87 74 27 / 80 - [email protected]
• Lung Infection and Innate Immunity
François TROTTEIN
• Lactic Acid Bacteria and Mucosal Immunity
Bruno POT
• Basic and Clinical Immunology of Parasitic Diseases
Sylviane PIED
• Pulmonary Immunity
Anne TSICOPOULOS
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ii) Strategies of Infection
A - Research Report
Team 3 : Plague and Yersinia pestis (Florent Sebbane)
Team 4 : Bacterial Respiratory Infections : Pertussis and
Tuberculosis (Camille Locht)
Team 5 : Cellular Microbiology of Infectious Pathogens
(Frank Lafont)
Team 6 : Molecular and Cellular Virology of Hepatitis C
(Jean Dubuisson)
Since its first days of existence, the Institut Pasteur de Lille has
always been very active in research on infectious diseases. It
has always combined basic aspects of science with potential
applications. The first major success was the development of
the bacille de Calmette et Guérin (BCG) by the first Director
General of the Institute, Albert Calmette, and his co-worker
Camille Guérin. BCG is the most widely used vaccine in the
world and is today still the only vaccine available against
tuberculosis and leprosy.
iii) Host Response and Inflammatory Processes
Team 7 : NODS-Like Receptors in Infection and Immunity
(Mathias Chamaillard)
Team 8 : Lung Infection and Innate Immunity (François
Trottein)
Team 9 : Lactic Acid Bacteria and Mucosal Immunity
(Bruno Pot)
Team 10 : Basic and Clinical Immunology of Parasitic
Diseases (Sylviane Pied)
Team 11 : Pulmonary Immunity (Anne Tsicopoulos)
Since then, a large body of research topics has evolved,
including a strong activity in parasitology, virology,
bacteriology and immunology.
The teams that are now part of this new project called «Center
for Infection and Immunity of Lille» (CIIL) have initiated and
have been actively involved in the organisation of important
events. These include the organisation of an international
symposium to celebrate the 150th birthday of microbiology
(December 10-12, 2007), the organization of an international
course on M. tuberculosis typing, based on the technologies
developed by one of the teams, the organisation of events
specifically addressing children (Kid-Campus). Furthermore,
several team heads or senior scientists are coordinators of
large European consortia in FP6 and in FP7.
Platform : Transcriptomics and Applied Genomics
(Yves Lemoine)
Some of these teams have already been functioning as
structured groups over the last 4 years (e. g. teams # 2, 3, 4, 5,
6, 9, 11), whereas others are newly constituted (e. g. teams #
1, 7, 10) and others are the result of PI movements between
previous units (e. g. team # 8). Many of the teams are headed
by young, promising investigators (at the senior CR or junior
DR level). Consequently, the teams vary quite substantially in
size. However, we believe that the sizes of the teams are
adapted to their scientific objectives.
The teams that are now part of the CIIL were previously
part of :
- Inserm U547, “Schistosomiase, Paludisme et Inflammation”
- Inserm U629, “Mécanismes Moléculaires de la Pathogénie
Bactérienne”
- Inserm U774, “Biomolécules et Inflammation Pulmonaire”
- Inserm U801, “Interactions Cellulaires et Moléculaires Hôte
- Pathogène”
- CNRS UMR 8161, “Institut de Biologie de Lille”
- EA2689, “Détresses Respiratoires et Circulatoires”
- EA3609, “Ecologie du Parasitisme”
The decision of this Center project is timely, since most units
finished their term by December 2009, and there are clear
conceptional and/or technological synergies across groups of
teams, which can be summarized in a number of common
themes :
- Transcriptional regulation of gene expression in pathogens
in response to environmental signals : in the different
models : Toxoplasma (MCBTG), Schistosoma (MBSDR),
Yersinia (PYP), Bordetella and Mycobacterium (BRIPT, TAG).
- Cellular biology of pathogens : in Plamodium (CMIP),
Toxoplasma (MCBTG), Schistosoma (MBSDR), Yersinia (PYP,
CMIP), Bordetella and Mycobacterium (BRIPT, CMIP), HCV
(MCVHV)
- Innate Immunity in infectious and non-infectious models :
in Yersinia (PYP, NLRII), Bordetella (BRIPT, LIII), probiotics (PYP,
NLRII, LABMI)
- Pulmonary immunity in infectious and non-infectious
diseases : in Bordetella (BRIPT, LIII), Mycobacterium (BRIPT, PI)
- Chronic inflammatory diseases : in the digestive tract (NLRII,
LIII, LABMI) in the respiratory tract (LIII, PI, BRIPT)
- Regulatory T cells (BRIPT, LIII, PI, BCIP)
- Vaccine development (BRIPT, LABMI, BCIP)
- Drug targets (MBSDR, BRIPT)
All these research units ended their first (U774, U801,
UMR8161) or second term (U547), except for Inserm U629, the
second term of which has started on January 1st, 2008.
A total of 11 teams and one platform have evolved from these
units. They are now structured into three major areas of
research, although there is no clear definition or distinction
between the three areas (e. g. it is clear that the strategies of
infection include some aspects of the biology of the pathogen
and some aspects of the host responses) : (i) Biology of the
Pathogen ; (ii) Strategies of Infection ; (iii) Host Responses and
Inflammatory Processes. Although several of these teams have
also strong cross-over activities with the different other teams,
one of them is mainly a cross-over team.
i) Biology of the Pathogen
Team 1 : Molecular and Cellular Biology of Toxoplasma gondii
(Stan Tomavo)
Team 2 : Molecular Biology of Schistosome Development and
Reproduction (Ray Pierce)
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Consequently, many interactions consisting of more or less
structured collaborations already exist between scientists of
the different teams. Some of them have been formalised and
have led to common publications or common grants. Many of
the teams have also been engaged in collaborations outside
of these 12 CIIL groups, either with regional laboratories,
within or outside of the IFR142, within France, Europe or
outside Europe, in particular with laboratories located in
developing countries, including the so-called emerging
countries, such as China (PI), India (BRIPT) and Brazil (BRIPT).
Strong links to Industry (BRIPT, LABMI, PI, CMIP, TAG) and to
the clinic (BRIPT, PI) have also been developed by several
teams. All these interactions will be further strengthened by
the creation of the CIIL.
Director for a four-year term, are taken on the basis of
consensus discussions or voting by the Executive Committee,
composed of all the team leaders. Excellence will be achieved
and maintained with the help of a high-level external
Scientific Advisory Board that is consulted before taking each
major strategic decision. Interactions between teams of the
CIIL are fostered by the identification of cross-over themes and
technologies, as well as by common seminars. A major
emphasis is put on interactions of the CIIL teams with the
outside world, in particular with top research centers in other
locations in France and elsewhere. Strong interactions already
exist with developing countries, in particular India, China,
Latin America and Africa.
B - Perspectives and
development 2010
Microbial and parasitic infections, as well as inflammatory
diseases are today still the causes of high rates of morbidity
and mortality world-wide. The most recent World Health
Report estimates that among the 50 to 60 million annual
human deaths roughly 1/4 succumb to infection. This
represents only the tip of the iceberg, in that the morbidity
and the long-term impact of infections on additional lifethreatening diseases, including chronic inflammatory
disorders, are likely to surpass by far this proportion. The
understanding of infection and immunity, including noninfectious immune-dysregulation, obviously requires a
multidisciplinary and integrated approach. Deciphering the
natural history of disease and the underlying mechanisms of
immune homeostasis and host defence may ultimately lead
to novel approaches for diagnosis, prognosis, treatment and
prevention by vaccines and/or immunomodulation.
The objectives of the Center for Infection and Immunity of Lille
(CIIL) lie precisely within this context. The CIIL is composed of
12 research teams of variable size, organized in three major
research areas, without clear distinction or delimitation
between them. It gathers complementary expertise, covering
a wide range of disciplines from epidemiology, over molecular
and cellular virology, bacteriology and parasitology, to the
immunological basis of infectious and non-infectious diseases
and translation into clinical applications. The CIIL will
make use of the most modern technologies, including
transcriptomics, proteomics, comparative and functional
genomics, structural biology, biophysics and cellular imaging.
The targeted diseases include some of the major infections
world-wide, such as hepatitis C, tuberculosis, malaria,
respiratory and gastro-intestinal infections. However,
other diseases of increasing public health concern are also
addressed,
including
plague,
whooping
cough,
schistosomiasis and toxoplasmosis. Furthermore, deciphering
the dialogue with the symbiotic microbiota might provide
novel clues in our understanding of chronic inflammatory
diseases, such as inflammatory bowel disease, asthma and
chronic obstructive pulmonary disease.
In order to accomplish the scientific objectives, a strong
management organisation is required. Important decisions,
including the election of a Director General and a Deputy
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Contents
of the biology of T. gondii. Our first aim is to identify and
characterize transcription factors and their functional
networks, with emphasis on the master factors required for
the coordinated expression of genes involved in the parasite
differentiation. Second, we propose to elucidate the molecular
bases of protein trafficking and post-Golgi secretory vesicle
formation involved in the biogenesis of secretory organelles
and the inner membrane complex that are only present in
apicomplexan parasites.
Molecular and cellular
biology of Toxoplasma
gondii
Stanislas TOMAVO, DR2 CNRS
Group Members :
Gissot Mathieu, CR2 CNRS
Delhaye Stéphane, IE CNRS (CDD)
Mouveaux Thomas, IE CNRS (CDD)
Hardy Sarah, Postdoctoral fellow
Walker A. Robert, Postdoctoral fellow
Fauquenoy Sylvain, PhD Student MENRT
Sloves Julien, PhD Student MENRT
Escobar-Ramirez Adelina, PhD Student Mexican fellowship
1. Understanding stage
differentiation in T. gondii
Despite the recent completion and annotation of several
apicomplexan genomes (http://www.toxodb.org), it is still
challenging to identify classical transcription factors of
apicomplexan parasites. We have established that the parasite
stage conversion is accompanied by the expression of a
variety of genes that display diverse functions such as parasite
motility, host recognition and invasion, parasite metabolism.
These observations strongly suggest that parasite
differentiation is regulated, at least in part, at the
transcriptional level. Therefore, we have decided to identify
the cis-acting elements and the transcription factors involved
in T. gondii gene regulation.
Key words :
Toxoplasma gondii. Parasite differentiation. Gene regulation.
Transcription factors.
A - Research Report
Mapping of sequence-specific promoter regions that bind to
putative transcription factors. We have shown that the
promoter regions of ENO1 and ENO2 display promoter
autonomy that can be exploited to follow developmental
expression of reporter genes. In addition, we have used a
combination of sensitive sequence analysis methods
(hydrophobic cluster analysis) in collaboration with Dr. Isabelle
Callebaut (CNRS UMR 7590, University of Jussieu, Paris 6-7) to
predict the existence of genes encoding several general
transcription factors in two apicomplexan parasite models,
Plasmodium falciparum and T. gondii. Taken together, the data
suggest that more transcriptional factors may be present in
the apicomplexa proteomes than initially thought.
The MCBTG team has emerged from a CNRS UMR 8576,
(Structural and Functional Glycobiology) located at the
campus of the University of Techonology of Lille. Since 1999,
the team has benefited from an ATIPE grant. This team has
now decided to join the CIIL project and will be located on the
campus of the Institut Pasteur de Lille early 2009. The team
has worked on the molecular and cellular biology of
Toxoplasma gondii since several years now.
T. gondii is an obligate intracellular protozoan parasite that is
a leading cause of focal central nervous system infections in
patients with AIDS/HIV. In addition, toxoplasmosis is also a
clinically important opportunistic pathogen during cancer
treatment, organ transplant and in newborns. Toxoplasma is
incurable, because of its ability to differentiate from the
rapidly replicating tachyzoite stages into latent cyst forms
containing the bradyzoite stages that is impervious to
immunity and current drugs. This developmental process is
triggered by the host immune response. Impairment of the
immune system in HIV-infected individuals can lead to stage
conversion of latent and avirulent bradyzoites into virulent
tachyzoites that cause lethal toxoplasmosis encephalitis. In
addition, apicomplexans have evolved an intricated and
sophisticated network of constitutive and regulated secretory
pathways, that is unparalleled in most eukaryotic cells. PostGolgi trafficking in apicomplexan parasites involves targeting
to three distinctive secretory organelles, named microneme,
rhoptry and denses granules, that play key roles in hostparasite interactions. In T. gondii, two key aspects of virulence
that underlie pathogenesis are parasite differentiation and
daughter-parasite formation, with parasite-specific organelle
biogenesis, both of which require changes in genomic
expression profile. Remarkably little is known about how
T. gondii determines the control of transcription and gene
expression that is central in managing the complex life cycle
of parasites, organelle biogenesis, parasite division and
differentiation. Our team aims to investigate these key aspects
Identifying the transcription factors binding to stage-specific
promoters of T. gondii. We have further analyzed the
genetically mapped intergenic region of the ENO1 gene,
which is transcriptionally repressed in tachyzoites, while it is
activated in bradyzoites. We have carried out an affinity
purification procedure for the nuclear factors that bind to the
450-bp promoter region described above. In collaboration
with the laboratory of Dr. Alain Van Dorsselaer (CNRS UMR
7178, University of Strasbourg), we have identified forty novel
factors by mass spectrometry, using a screening of the whole
genome sequences. Three putative transcription factors
encoded by genes that are exclusively expressed in
tachyzoites were selected. These three proteins have no
sequence similaroty to any factors known in other organisms,
suggesting that these factors are exclusively present in T.
gondii and other apicomplexan parasites. We have
demonstrated that the purified recombinant factors can
specifically bind to known DNA targets of 45-bp nucleotides
in length. In addition, we have confirmed their nuclear
localization, their specific binding to their cognate promoter
in vivo, using the polyclonal antibodies and chromatin
immuno-precipitation. The biological functions of these
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putative transcription factors will be further elucidated in
more detail.
initially thought. Moreover, our studies also revealed that
these novel vesicles contain a sortilin-like N-glycoprotein,
which belongs to a growing family of multi-ligand type-1
membrane receptors that are involved in lysosomal trafficking.
We have found that the T. gondii sortilin-like protein is mainly
localized in the Golgi compartment of the parasite, in addition
to the novel unkown vesicles described above. However, only
the upper compartment of the Golgi was labelled, suggesting
that sortilin is present in the trans-Golgi network and/or in the
post-Golgi compartment. Because the homologue of a cationindependent mannose 6-phosphate receptor has not been
described in any apicomplexan parasites, we postulate that
the T. gondii sortilins are likely to play key roles as alternative
protein-sorting receptors in T gondii. The biological functions
of sortilin will be further elucidated in more detail.
AP2 domain family of proteins : key transcriptional regulators
for the Apicomplexa ?
The genome of T. gondii encodes around 45 genes with one
or more copies of the AP2 DNA binding domain. Because of
their origin in the plant lineage, these AP2 proteins have no
homologues in the human host and may prove to be ideal
novel anti-apicomplexan drug targets. We have confirmed the
stage-specific expression of several putative AP2 transcription
factors. Some of the AP2 genes have been knock-in and their
biological functions are under investigation.
2. Organelle Biogenesis
and Differentiation in T. gondii
development 2010
The T. gondii cell is relatively small (2x8 m) banana-shaped,
and the parasite’s architecture can be appreciated in a few
electron microscope thin sections, showing that it displays a
single nucleus, a single mitochondrion, a single plastid, a
single interconnected ER network, a single Golgi apparatus,
and an apical clustered complex of secretory organelles,
exclusively found in apicomplexans. Toxoplasma provides a
convenient bridge for understanding the architecture of even
more highly simplified organisms with the ease to interpret in
morphological terms, and for which genetic experiments are
also feasible. All stages of T. gondii and those of other
apicomplexan parasites are polarized cells that contain
unusual organelles, such as rhoptries, micronemes and denses
granules, that are involved in host cell invasion. Post-Golgi
trafficking in apicomplexan parasites involves targeting to
these three distinctive secretory organelles. Little is known
about membrane proteins that mediate the membrane
transport during the biogenesis of the specialized secretory
organelles and the inner membrane complex in apicomplexan
parasites. Our second aim is to investigate the molecular bases
of protein trafficking and post-Golgi secretory vesicle
formation involved in the biogenesis of T. gondii secretory
organelles.
Our experimental model T. gondii, like other apicomplexan
parasites, displays unique apical secretory organelles
mediating locomotion and cellular invasion. We now have
sufficient knowledge of global mRNA expression in
Toxoplasma to conclude that transcriptional mechanisms play
a major role in regulating the developmental program of the
parasite. The observations that co-regulated genes are
dispersed across parasite chromosomes, along with the
presence of cis-elements suggest that promoters of T. gondii
are regulated by trans-acting factors, which remain to be
identified and characterized. Our first specific aim is to identify
and characterize the transcription repressors and activators of
the parasite. In addition, our second aim will provide a cellular
and an in vivo functional assay system, which will be helpful
to decipher vesicle shuttle and protein trafficking in T. gondii.
In the future, we will identify and characterize the master
transcriptional regulators involved in the coordinated
expression of genes involved in organelles biogenesis during
formation of daughter parasites. The data collected here can
be exploited to other apicomplexan parasites and, this
certainly represents a great challenge that may yield new
insights into the biology of these protozoan parasites.
Identification and characterization of novel post-Golgi
vesicles. During the course of our proteomics studies, we have
identified homologues of Cathepsin C proteases, which are
known to play key roles in apicomplexan invasion, organellar
biogenesis and intracellular survival. In collaboration with Dr.
Sharon Reed (University of California, San Diego, USA), we
have characterized two genes encoding the cathepsins,
named TgCPC1 and TgCPC2, respectively. We found that these
genes are exclusively expressed in the tachyzoites, and they
are surprisingly localized in the dense granules. We have
demonstrated that these enzymes are key enzymes for
intracellular development of the parasites using reverse
genetics and biochemical (specific inhibitors) approaches. The
cathepsin Cs and other enzymes can represent excellent tools
as powerful biochemical tracer of protein sorting during
organelle biogenesis, as well as N-glycosylation, a posttranslational protein modification occurring in the
endoplasmic reticulum-Golgi trafficking. Previously, it was
only assumed that limited N-linked glycosylation exists in
apicomplexan parasites, until we recently demonstrated that
N-glycosylation is much more prevalent in T. gondii than
Publications
2007
Daher W, Oria G, Fauquenoy S, Cailliau K, Browaeys E, Tomavo S,
Khalife J.
A Toxoplasma gondii leucine rich repeat protein binds phosphatase type
1 protein and negatively regulates its activity.
Eukaryot Cell, 2007, 6:1606-1617.
Gissot M, Kelly KA, Ajioka JW, Greally JM, Kim K.
Epigenomic Modifications Predict Active Promoters and Gene Structure
in Toxoplasma gondii.
PLoS Pathog, 2007, 3:e77.
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Que X, Engel JC, Ferguson D, Wunderlich A, Tomavo S, Reed SL
Cathepsin Cs are key for the intracellular survival of the protozoan
parasite, Toxoplasma gondii.
J Biol Chem, 2007, 282 :4994-5003.
PhD
MEURICE Edwige
Directeur de thèse : Stanislas TOMAVO
Caractérisation de deux facteurs transcriptionnels putatifs de
Plasmodium falciparum, PfTAF7 et PfTAF10
Université des Sciences et Technologie de Lille
15 Octobre 2007
Asai T and Tomavo S.
Biochemistry and metabolism of Toxoplasma gondii. In “Toxoplasma
gondii : The model apicomplexan-Perspectives and Methods”.
Edited by Weiss, L.M. & Kim. K. Academic Press, Elsevier, London. UK,
2007, p 185-206.
Tomavo S and Weiss L.M.
Toxoplasma gene regulation and bradyzoite development. In “The
biology of Toxoplasma gondii”.
Edited by Soldati, D. & Ajioka, J. Horizon Press Book, UK, 2007, p 285-301.
2008
Deschamps P, Guillebeault D, Devassine J, Dauvillée D, Haebel S,
Steup M, Buléon A, Putaux JL, Slomianny MC, Colleoni C, Devin A,
Plancke C, Tomavo S, d’Herelle E, Moreau H, Ball S.
The heterotrophic dinoflagellate Crypthecodinium cohnii defines a model
genetic system to investigate cytoplasmic starch synthesis.
Eukaryot Cell, 2008, 7:872-880.
Fauquenoy S, Morelle W, Hovasse A, Bednarczyk A, Slomianny C,
Schaeffer C, Van Dorsselaer A, Tomavo S.
Proteomics and glycomics analyses of N-glycosylated structures involved
in Toxoplasma gondii-host cell interactions.
Mol Cell Proteomics, 2008, 7:891-910.
Gissot M, Choi SW, Thompson RF, Greally JM, Kim K.
Toxoplasma gondii and Cryptosporidium parvum lack detectable DNA
cytosine methylation.
Eukaryot Cell, 2008, 7:537-540.
Gissot M, Ting LM, Daly T, Bergmann LW, Voza T, Sinnis P, Kim K.
High mobility group protein HMGB2 is a critical regulator of plasmodium
oocyst development.
J Biol Chem, 2008, 283:17030-17038.
Thompson RF, Reimers M, Khulan B, Gissot M, Chen Q, Zheng X, Kim
K, Greally JM.
An analytical pipeline for genomic representations used for cytosine
methylation studies.
Bioinformatics, 2008, 24:1161-1167.
Ting LM, Gissot M, Coppi A, Schramm V, Sinnis P, Kim K.
Attenuated Plasmodium yoelii Lacking Purine Nucleoside Phosphorylase
Confer Protective Immunity.
Nat Med, 2008, 14:954-958.
2009
Gissot M, Kim K, Schaap D, Ajioka JW.
New eukaryotic systematics : a phylogenetic perspective of
developmental gene expression in the Apicomplexa.
Int J Parasitol, 2009, 39:145-151.
2010
Dauvillée D, Delhaye S, Gruyer S, Slomianny C, Moretz SE,
d’Hulst C, Long CA, Ball SG, Tomavo S.
Engineering the chloroplast targeted malarial vaccine antigens in
Chlamydomonas starch granules.
PLoS One, 2010, In press.
Holmes M, Liwak U, Pricop I, Wang X, Tomavo S, Ananvoranich S.
Silencing of tachyzoite enolase 2 alters nuclear targeting of bradyzoite
enolase 1 in Toxoplasma gondii.
Microbes Infect, 2010, 12:19-27.
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Contents
receptor for growth factors or hormones directing signalling
pathways potentially involved in these processes; membrane
receptor tyrosine kinases and nuclear hormone receptors.
Both types of receptor could potentially interact with
endogenous or exogenous (host) ligands in order to regulate
development and differentiation in our study model
Schistosoma mansoni.
Molecular biology
of Schistosome
development
and reproduction
Raymond PIERCE, DR2 CNRS
1. Kinase activities in parasite
development and reproduction
Group Members :
Dissous Colette, DR2 Inserm
Dive Daniel, DR2 Inserm
Khalife Jamal, DR CNRS
Pierrot Christine, CR IPL
Caby-Coutte Stéphanie, IE IPL
Debreucq Nadège, Technician IPL
Godin Claude, Technician IPL
Kalamou-Mama Hadidjatou, Technician IPL
Lafitte-Carton Sophia, Technician IPL
Trolet Jacques, Technician IPL
Dubois-Abdesselem Florence, Postdoctoral fellow MENRT
Fréville Aline, PhD student MENRT
Gouignard Nadège, PhD student MENRT
Lancelot Julien, PhD student Univ Lille 2
Vanderstraete Mathieu, PhD student Univ Lille 2
Vandomme Audrey, PhD student Univ Lille 2
Reversible protein phosphorylation regulates most of the
basic cellular functions and energy metabolism in all
eukaryotes. In metazoan organisms, protein phosphorylation
also coordinates cell and organ differentiation, as well as
communication between cells or with the external
environment. Receptor Tyrosine Kinases (RTK) and their
kinase-associated pathways regulate the effects of growth
factor signalling on metabolism, growth and differentiation.
We initially targeted two RTK subfamilies, the ligands of which,
epidermal growth factor (EGF) and insulin, modulate
schistosome metabolism.
Although these receptors present structural and functional
similarities with their host counterparts, comparative threedimensional modelling of their ATP-binding sites indicated
that structural divergences with host receptors exist that
would be exploitable for the design of kinase inhibitors
specific for schistosome RTKs.
More importantly, we also discovered a novel RTK in
S. mansoni that belongs to a hitherto unknown class with no
counterpart in humans. This novel receptor (SmVKR for Venus
Kinase Receptor) is formed by an IR-like tyrosine kinase
domain coupled to an extracellular module named Venus
Flytrap, usually found in neurotransmitter receptors and some
sensory receptors. Its preferential localization in schistosome
germinal cells suggests that it could be involved in parasite
reproduction, and thereby could represent an excellent target
for reducing both pathology and transmission of
schistosomiasis. We have shown that SmVKR belongs to a
novel family of RTKs found only in invertebrates and
preferentially in insects. SmVKR have the conserved properties
of RTK such as the capacity to homodimerize and to
autophosphorylate in the presence of a ligand (Ahier et al,
2009).
In addition, we have characterized two polo-like kinases
(SmPlk1 and SmSak) associated with the reproductive organs
of schistosomes and their key functions in cell division have
been demonstrated. The Plk1 inhibitor (BI2536) provokes
important morphological alterations of the schistosome
reproductive organs, indicating the importance of SmPlk1 as
a possible target for chemotherapy(Long et al, submitted).
Moreover, we have characterized an Ste20-like protein kinase
(SmSLK), related to the “germinal center kinases” that
possesses a polo-like kinase kinase (plkk) activity. We have
recently demonstrated that SmSLK activates SmPlk1 and
initiates G2/M transition during mitosis, in a caspasedependent manner. This upstream kinase of SmPlk1 is another
potential chemotherapeutic target.
Key words :
Schistosoma mansoni. Kinase signalling. Gametogenesis.
Cell division. Receptor tyrosine kinase. Epigenetics. Histone
deacetylase. Drug discovery. Vaccine. Plasmodium falciparum.
Immunity. Phosphatase.
A - Research Report
The MBSDR team has emerged from Inserm U547 within
which Drs. Dissous and Pierce were responsible for the
Schistosoma molecular biology programme, with the main
emphasis on receptor-mediated signalling and transcriptional
control. Recently, we have been joined by the group led by Dr.
J. Khalife whose activities cover both schistosomiasis vaccine
development and themes concerning the malaria parasite,
Plasmodium sp.
Platyhelminth parasites of the genus Schistosoma infect 200
million individuals worldwide and cause more than 200,000
deaths annually, mainly in sub-Saharan Africa. Moreover, until
very recently, the impact of schistosomiasis on infected
populations was drastically underestimated. Only one
effective drug, praziquantel, exists for the treatment of the
disease, meaning that the selection of resistant parasite strains
is probable. The discovery and development of new
schistosomicidal drugs is a “Strategic Emphasis” of the WHO.
Schistosomes have a complex life-cycle involving two hosts,
two types of reproduction and four morphologically distinct
stages. The strategy of our group is to develop a better
understanding of the mechanisms involved in schistosome
development and reproduction, in order to devise novel tools
for the control of the parasite. We targeted two kinds of
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Contents
These latter findings and the methodologies put in place
prepared the way for our current investigation of the role of
epigenetic mechanisms, specifically involving histone
modifications, in the control of transcription during
schistosome development. This project, carried out in
collaboration with the group of G. Mitta (CNRS UMR 5244,
Perpignan) and supported by a grant from the ANR, focuses on
genes coding for mucin-like glycoproteins the expression of
which is tightly controlled during the S. mansoni life-cycle.
Moreover, our finding that HDACi can be toxic to schistosomes
opens up the possibility novel drug strategies against the
parasite. A project, coordinated by us and involving 8 european
and brazilian partners with the aim of developing novel
inhibitors of schistosome histone-modifying enzymes as drugs
(SEtTReND) has been funded for 3 years by the EC (FP7-Health).
2. From transcription factors
to epigenetics
During metazoan development, the control of gene
expression implies precise coordination between the
activation and the repression of sets of genes. These
mechanisms involve numerous DNA-binding transcription
factors, and in particular a superfamily of ligand-regulated
transcription factors, the nuclear receptors. We focused our
studies on the S. mansoni nuclear receptor Ftz-F1. Although
the Ftz-F1 family members are monomeric orphan receptors,
we showed that SmFtz-F1 possesses novel functional
properties, including specific heterodimerization with
SmRXR1 (S. mansoni retinoid X receptor 1) and we also
suggested the possibility of ligand-dependent activity. The
original functional properties of SmFtz-F1 suggested that this
nuclear receptor might also interact with novel protein
partners.
Nuclear receptors regulate target gene expression by binding
to their cognate DNA response elements and recruiting
associate protein cofactors, which either activate or repress their
transcriptional activity. The cofactors can be classified in two
groups, the coactivators and the corepressors, neither of which
possess intrinsic DNA binding properties. These proteins
function either directly by remodelling chromatin structure
and/or by acting as adaptor molecules in multi-subunit protein
complexes. Transcriptional activation is usually associated with
chromatin remodelling carried out in particular by the
coactivators possessing HAT (histone acetyl transferase)
enzyme activity. This modification of the histone “opens” the
chromatin structure and allows transcription of the target gene.
We characterized two highly conserved representatives of the
CBP/p300 family of transcriptional coactivators, and
demonstrated functional interaction with SmFtz-F1.
Conversely, when the nuclear receptors are not activated, they
interact with corepressors. In contrast to nuclear receptor
coactivators, corepressors do not possess any intrinsic HDAC
(histone deacetylase) activity, but can recruit an HDAC complex,
which deacetylates histones and “compacts” chromatin,
resulting in transcriptional repression. We characterized a
completely novel corepressor of SmFtz-F1 termed SmFIP-1
(SmFtz-F1 interacting protein-1). SmFIP-1 interacted both
physically and functionally with SmFtz-F1, repressing its
transcriptional activity. No homologues of SmFIP-1 have been
found outside the Schistosoma genus, suggesting that some
transcriptional control mechanisms involving SmFtz-F1 may be
specific to schistosomes and form part of an adaptation to the
parasitic way of life. We subsequently cloned and characterized
all three class 1 HDACs present in the genome, orthologues of
mammalian HDACs 1, 3 and 8. They have conserved catalytic
domains, but divergent C-terminal domains. Moreover, in order
to determine the extent and importance of histone acetylation
in S. mansoni, we tested the effects of histone deacetylase
inhibitors (HDACi) on both larval and adult worms in culture.
Trichostatin A (TSA) caused dose-dependent mortality of
schistosomula and adults and an increase in general levels of
histone acetylation in schistosomes, particularly of histone H4.
Increased H4 acetylation was accompanied by the increased
expression of HDAC target genes, including caspase 7, and this
increase correlated with increased H4 acetylation of the
caspase 7 gene promoter, measured by quantitative chromatin
immunoprecipitation.
Development 2010
Schistosomes have a complex life-cycle, involving two hosts,
two modes of reproduction and four morphologically distinct
forms. The success of their development is dependent on their
adaptation to the host environment and receipt of host signals
and on their ability to control strictly the expression of genes
key to their developmental programmes. The strategy of our
group is to develop a better understanding of the
mechanisms involved in schistosome development and
reproduction in order to devise novel tools for the control of
the parasite. Our research projects follow on from the themes
developed over the last four years and focus on two main
areas, both intrinsic to host-parasite interactions determining
parasite development and differentiation: 1) signalling in
protein interaction and phosphorylation cascades and 2)
epigenetic mechanisms in the control of gene expression
1. Kinase signalling in schistosome
reproduction (C. Dissous)
Transmission and human pathology in schistosomiasis are due
for a large part to the exceptional fecundity of female worms.
Our projects concern the study of kinases with a well-known
pivotal role in cell division (polo-like kinases and upstream
regulators) and their function in vitellogenesis and oocyte
maturation. They also concern the functional characterization
of SmVKR, a receptor tyrosine kinase (RTK) potentially involved
in female maturation and reproductive activities.
Role of Venus kinase receptors (VKR) in invertebrate
fecundity
We have recently shown that the Venus Kinase Receptors
(VKR) that contain a Venus flytrap ligand domain and an
insulin receptor-like intracellular kinase domain constitute a
new family of RTK present in schistosomes but also in other
invertebrates, such as Anopheles gambiae, the malaria vector.
In S. mansoni and in A. gambiae, VKR are present in larvae and
female gonads, thus likely involved in development and
sexual differentiation. Structural and functional studies of VKR
could contribute to a better knowledge of development and
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Contents
reproduction within invertebrates. Since VKR are not found in
vertebrates, they could lead towards novel approaches in the
control of transmission and spreading of the two major
parasitic diseases in the world: schistosomiasis and malaria.
VKR are a new family of orphan receptors with unknown
functions.
Our project aims to explore the biological function of VKR in
S. mansoni and in A. gambiae by studying the regulation of
their expression during the development and differentiation
of reproductive organs and the consequences of VKR
invalidation on the fertility of females. Functional assays based
on the expression of VKR (wild-type and mutated, including
active and inactive constitutive versions) in different pertinent
cellular models (insect cells, mammalian cells and Xenopus
oocytes) will be set up to determine the functional properties,
signalling pathways and potential role in development of this
new family of RTK. Besides, we will analyse VKR activation
mechanisms by using chimera constructs formed by extra and
intracellular modules with well-known properties (mGluR and
c-MET respectively) combined with VKR modules. These
chimeras will allow the identification of ligands (natural or
synthetic) for VKR (collaboration COSMIC- UMR8161 CNRS).
2. Transcriptional control of
schistosome development (R. Pierce, S. Caby)
The generation of new genetic networks, together with the
diversification of the regulation of gene expression are central
mechanisms by which animal evolution has occurred. Whilst
schistosomes cannot be considered as “model” organisms, the
study of the transcriptional control mechanisms involved in
development and differentiation of these parasites will
advance knowledge of their evolution in eukaryotes.
Epigenetic mechanisms of transcriptional control in
S. mansoni : towards new drugs.
Taken together, our work over the past few years shows that
schistosomes conserve the major mechanisms of
transcriptional control present in other metazoans, but have
evolved functions and protein components of these
mechanisms that are schistosome-specific. The next phase will
involve pushing beyond the study of global histone
acetylation to a more detailed investigation of specific histone
marks involved in gene activation/silencing in S. mansoni. In
collaboration with G. Mitta (CNRS UMR 5244, Perpignan) in a
study financed by the ANR (project: Schistophepigen) we will
investigate the control of the expression of two gene families
coding for mucin-like glycoproteins with VNTR domains that
have important roles in host-schistosome interaction. The
tightly controlled differential expression of these two gene
families provides an excellent model for the investigation of
the specific epigenetic marks and protein partners involved
in this process.
The main emphasis of our work during the next three years
will concern the project funded by the EC (SEtTReND). In
collaboration with five European and three Brazilian partners
we will characterize and clone most of the enzymes involved
in histone acetylation/deacetylation and methylation/
demethylation. Promising candidates will be expressed as
recombinant proteins and the structures of their catalytic
domains will be determined by crystallography or in silico
modelling. Phenotypic profiling using RNAi and generic
inhibitors of histone modifying enzymes will validate potential
targets and generate gene expression profiles that will define
specific inhibition. Inhibitors will be validated both in silico and
in vitro and optimised drug candidates will be tested in vivo.
Overall the SEtTReND partners expect to generate several
candidate schistosomicidal drugs that will go forward for
further testing.
Role of SmSLK and Plks in gametogenesis in S. mansoni
The unexpected observation of the presence of the Ste20-like
kinase SmSLK in schistosome oocytes and its homology to a
Xenopus polo-like kinase kinase (xPlkk) indicated that it might
have a role in G2/M cell cycle regulation. We have shown that
the SmSLK kinase domain, devoid of the C-terminal coiled-coil
domains activates oocyte meiosis and that caspase activation
in oocytes promotes the cleavage of native SmSLK also
leading to the resumption of the cell cycle.
The project consists in the characterization of the role of
SmSLK in the activation of SmPlk1, and in the proliferation of
schistosome oocytes. Overall, this project aims to demonstrate
a novel regulatory role for caspases in the cell cycle that could
activate one of the essential effectors of the M phase.
The characterization of SmSak, a second Plk family member
homologous to Sak in Drosophila and associated to testis
formation in this organism, will be also performed. Its role in
mitosis and its possible activation by SmSLK will be studied in
the Xenopus oocyte model in collaboration with K. Cailliau
and E. Browaeys (Université de Lille 1).
Schistosome guanidino kinase (SmGK) and specific
inhibitors for the control of schistosome infection.
SmGK belongs to the creatine kinase family, i. e. a group of
enzymes essential for ATP and energy production. SmGK is
abundantly expressed in schistosome infective larvae
(cercariae, schistosomula) and therefore represents an
excellent target for the development of novel infection control
strategies, by allowing preferential killing of young parasites
against which the only currently available drug, praziquantel,
has no effect. In collaboration with J.M. Lancelin and O.
Marcillat (ESCPE, UMR CNRS 5013, Lyon) and in a study
financed by ANR (SmGKinhibit), we will develop specific
inhibitors for SmGK and will evaluate their protective effect in
rodent experimental infections.
3. Immunobiology of Parastic
diseases (C. Pierrot, D. Dive and J. Khalife)
Malaria is still the most devastating parasitic disease in terms
of public health and economic impact. In Plasmodium
falciparum, a considerable body of work based on immunoepidemiological field studies suggested that natural-acquired
immunity (called semi-immunity) protects hosts by limiting
the rate at which the parasite replicates within the infected
host. To date, however, it appears that there is a constant
competition between an adequate host immune response
that takes time to develop and parasite growth and
differentiation essential for antigenic shifts to escape these
responses. This pathogen seems to be obsessed by a rapid
18
Contents
Daher W, G. Oria G, Fauquenoy S, Cailliau K, Browaeys E,Tomavo S,
Khalife J.
A Toxoplasma gondii leucine-rich repeat protein binds phosphatase type
1 protein and negatively regulates its activity.
Eukaryot Cell, 2007, 6:1606-1617.
replication to stay ahead of immune responses in order to
ensure its transmission.
The first part of the project examines potential components
involved in signalling pathways of parasite replication. Indeed,
given the ‘atypical’ division of Plasmodium falciparum where
DNA replicates more than once per cell cycle, efforts have
begun for an extensive studies of kinases and phosphatases,
known for their role in cell cycle and division. In the case of
phosphatases, and particularly Protein Phosphatase type 1
(PP1), it is well described that the spatio-temporal activity of the
catalytic subunit is highly controlled by regulatory proteins.
Currently, we are investigating the role of conserved as well as
new regulators of the nuclear functions of PP1 in P. falciparum.
These regulatory subunits were identified by 1) similartity with
known proteins described in other eukaryotes for their capacity
to regulate PP1, 2) the presence of the degenerate RVXF
binding motif in their amino acid sequences and 3) their
capacity of to bind PfPP1 in vitro. The functional roles of these
regulatory proteins were characterized by biochemical
approaches and by genetic analysis. These studies will provide
a basic framework to understand the exact role of the different
regulatory proteins of PP1 in the biology of Plasmodium.
The second part of the project is devoted to decipher how the
control of blood stage parasite multiplication can be achieved
by the host defences by taking into account the age of the host.
Indeed, it has repeatedly been observed in endemic areas,
mainly in Africa, that the most susceptible population are
children under the age of 5 years. We have investigated this
phenomenon by the establishment of a rat experimental model
of infection with P. Berghei. In this model, we observed that
young rats (< 4 weeks of age) developed high parasitemia and
succumbed to infection, while adult rats (> 8 weeks of age)
controlled their blood parasitemia and survived thereafter. This
model is now used to dissect the cellular and molecular
mechanisms that are involved in either the expression of
susceptibility or protection related to the age of the host.
Finally, based on our long-term experience with the rat model
in parasitic infections and on our knowledge of the rat
immune responses showing a certain similarity with those
induced in infected humans by Schistosoma mansoni, we
participate in a targeted development of new generation
vaccine for schistosomiasis. This research project, which is a
part of an international consortium granted by FP7 for the
next 4 years, will use the rat model to evaluate the immune
responses as well as the efficacy of novel vaccine candidates
against schistosomiasis.
Dissous C, Ahier A, Khayath N.
Protein tyrosine kinases as new potential targets against human
schistosomiasis.
Bioessays, 2007 29:1281-1218.
Duboucher C, Caby S, Chabé M, Gantois N, Delgado-Viscogliosi P,
Pierce RJ, Capron M, Dei-Cas E, Viscogliosi E.
Les trichomonoses pulmonaires humaines.
Presse Med, 2007, 36:835-839.
Duboucher C, Barbier C, Beltramini A, Rona M, Ricome JL, Morel G,
Capron M, Pierce RJ, Dei-Cas E, Viscogliosi E.
Frequent pulmonary superinfection by trichomonads in the course of
acute respiratory distress syndrome.
Lung, 2007, 185:295-301.
Duboucher C, Boggia R, Morel G, Capron M, Pierce RJ, Dei-Cas E,
Viscogliosi E.
Pneumocystis pneumonia: immunodepression, Pneumocystis jirovecii …
and the third man.
Nat Rev Microbiol, 2007, 5 (12).
Dufernez F, Villanueva MR, Noël C, Caby S, Delgado-Viscogliosi P,
Ohkuma M, Kudo T, Capron M, Pierce R, Walker RL, Viscogliosi E.
Morphological and molecular identification of non-Tritrichomonas fœtus
trichomonad protozoa from the bovine preputial cavity.
J Eukaryot Microbiol, 2007, 54:161-168.
Khayath N, Vicogne J, Ahier A, Ben Younes A, Konrad C, Trolet J,
Viscogliosi E, Brehm K, Dissous C.
Diversification of the insulin receptor family in the helminth parasite
FEBS J, 2007, 274:659-676.
Ludolf F, Bahia D, Cousin A, Capron M, Pierce R, Dissous C, Oliveira G.
Molecular analysis of SmFes, a novel tyrosine kinase of Schistosoma
mansoni orthologous to the members of the Fes/Fps/Fer family.
Biochem Biophys Res Commun, 2007, 360:163-172.
Noël C, Noda S, Mantini C, Dolan MF, Moriya S, Delgado-Viscogliosi P,
Kudo T, Capron M, Pierce RJ, Ohkuma M, Viscogliosi E.
Molecular Phylogenetic position of the genera Stephanonympha and
Caduceia (Parabasalia) inferred from nuclear small subunit rRNA gene
sequences.
Pierrot, C., E. Adam, D. Hot, S. Lafitte, M. Capron, J. D. George, and
J. Khalife.
Contribution of T cells and neutrophils in protection of young susceptible
rats from fatal experimental malaria.
J Immunol, 2007, 178:1713-1722.
Publications
Vanbesien-Mailliot CC, Wolowczuk I, Mairesse J, Viltart O, M.
Delacre M, Khalife J, Chartier-Harlin MC, Maccari S.
Prenatal stress has pro-inflammatory consequences on the immune
system in adult rats.
Psychoneuroendocrinol, 2007, 32:114-124.
2007
Bahia D, Mortara R, Freire L, Ludolf F, Kuser PR, Avelar L, Pierce R,
Dissous C, Oliveira G.
Schistosoma mansoni : Expression of Fes-like tyrosine kinase SmFes in the
oral sucker and terebratorium suggests its involvement in host
penetration.
Exp Parasitol, 2007, 116:225-232.
Yan YT, Tulasne D, Browaeys E, Caillau K, Khayath N, Trolet J, Pierce
RJ, Fafeur V, Ben Younes A, Dissous C.
Molecular cloning and characterization of SmSLK, a novel Ste20-like
kinase in the human parasite Schistosoma mansoni.
Int J Parasitol, 2007 37:1539-1550.
Daher W, Pierce RJ, Khalife J.
Census, molecular characterization and developmental expression of
Leucine-Rich-Repeat proteins in Plasmodium falciparum.
Mol Biochem Parasitol, 2007, 155:161-166.
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Contents
Berriman M, Haas B, Loverde PT, Wilson RA, Dillon GP, Cerqueira GC,
Mashiyama ST, Al-Lazikani B, Andrade LF, Ashton PD, Aslett MA,
Bartholomeu DC, Blandin G, Caffrey CR, Coghlan A, Day TA,
Delcher A, De Marco R, Djikeng A, Eyre T, Gamble JA, Ghedin E, Gu Y,
Hertz-Fowler C, Hirai H, Hirai Y, Houston R, Ivens A, Johnston D,
Lacerda D, Macedo CD, Mcveigh P, Ning Z, Oliveira G, Overington GP,
Parkhill J, Pertea M, Pierce RJ, Protasio AV, Quail MA, Rajandream M-A,
Rogers J, Sajid, M., Salzberg, S.L., Stanke, M., Tivey, A.R., White, O.,
Williams, D.L., Wortman, J., Wu, W., Zamanian M, Zerlotini A, FraserLiggett CM, Barrell BG, El-Sayed NM.
The Genome of the blood fluke Schistosoma mansoni.
Nature, 2009, 462:352-358.
2008
Ahier A, Khayath N, Vicogne J, Dissous C.
Insulin receptor and glucose uptake in the human parasite Schistosoma
mansoni.
Parasite, 2008, 15:573-579.
Awama AM, Paracuellos P, Laurent S, Dissous C, Marcillat O, Gouet P.
Crystallisation and X-ray analysis of the Schistosoma mansoni guanidino
kinase.
Acta Cryst, 2008, F64:854-857.
Chavain N, Vezin H, Dive D, Touati N, Paul JF, Buisine E, Biot C. 2008.
Investigation of the Redox Behavior of Ferroquine, a New Antimalarial.
Mol Pharm, 2008, 5:710-6.
Caby S, Pierce RJ.
Quantitative chromatin immunoprecipitation (Q-ChIP) applied to
Dive D, Biot C.
Ferrocene conjugates of chloroquine and other antimalarials : the
development of ferroquine, a new antimalarial.
Chem Med Chem, 2008, 3:383-391.
Dissous C, Ahier A, Long T.
A new promising drug against schistosomiasis.
Med Sci, 2009, 25:24-26.
Dubar F, Khalife J, Brocard J, Dive D, Biot C.
Ferroquine, an ingenious antimalarial drug : thoughts on the mechanism
of action.
Molecules, 2008, 13:2900-2907.
Dubar F, Anquetin G, Pradines B, Dive D, Khalife J, Biot C.
Enhancement of the antimalarial activity of ciprofloxacin using a double
prodrug/bioorganometallic approach.
J Med Chem, 2009, 52:7954-7957.
Duboucher C, Pierce RJ, Capron M, Dei Cas E, Viscogliosi E.
Recent advances in pulmonary trichomonosis.
Trends Parasitol, 2008, 204:201-202.
Dubois F, Caby S, Oger F, Cosseau C, Capron M, Grunau C, Dissous
C, Pierce RJ.
Histone deacetylase inhibitors induce apoptosis, histone hyperacetylation
and up-regulation of gene transcription in Schistosoma mansoni.
Dufernez F, Caby S, Derelle E, Noël C, Wyckmans J, Dive D, SoyerGobillard MO, Capron M, Pierce RJ, Wintjens R, Guillebault D,
Viscogliosi E.
Molecular characterization of iron-containing superoxide dismutases in
the heterotrophic dinoflagellate Crypthecodinium cohnii.
Protist, 2008, 159:223-238 .
Jiménez JC, Pinon A, Dive D, Grzych JM, Capron M, Di Prisco MC,
Dei-Cas E.
Specific IgA antibody response in children infected with Giardia
intestinalis after treatment with Secnidazol.
Am J Trop Med Hyg, 2009, 80:11-15.
Roger E, Gourbal B, Grunau C, Pierce RJ, Galinier R, Mitta G.
Expression analysis of highly polymorphic mucin proteins (SmPoMuc)
from the parasite Schistosoma mansoni.
Pierce RJ, Mitta G, Roger E.
Les génomes des schistosomes : une étape clé dans la lutte contre la
bilharziose.
Med Sci, 2009, 25:763-765.
Roger E, Grunau C, Pierce RJ, Hirai H, Gourbal B, Galinier R,
Emans R, Cosseau C, Mitta G.
Controlled chaos of antigenic variants in a metazoan parasite
(Schistosoma mansoni) interacting with its invertebrate host
(Biomphalaria glabrata).
PLoS Neg Dis, 2008, 2: e330.
Quack T, Knobloch J, Beckmann S, Vicogne J, Dissous C, Grevelding CG.
The formin-homology protein SmDia interacts with the Src kinase SmTK
and the GTPase SmRho1 in the gonads of Schistosoma mansoni.
PloS One, 2009, 4:e6998.
2009
2010
Amancha PK, Dissous C, Nadimpalli SK.
Characterization of the mannose 6-phosphate receptor (Mr 300kDa)
protein dependent pathway of lysosomal enzyme targeting in
Biomphalaria glabrata mollusc cells.
Biochimie, 2009, 8:982-988.
Acroute dit Vampouille A, Lafitte S, Dive D, Khalife J, Pierrot C.
A role for CD4+ and CD8+ cells and not for CD25+ cells in the control of
Plasmodium berghei Anka blood stage parasites in rats.
Parasite, 2010, 17:58-60.
Arancibia R, Dubar F, Pradines B, Forfar I, Dive D, Hugo Klahn A,
Biot C.
Synthesis and antimalarial activities of rhenium bioorganometallics
based on 4-aminoquinoline structure.
Bioorg Med Chem, 2010, in press.
Ahier A, Rondard P, Gouignard N, Khayath N, Huang S, Trolet J,
Donoghue DJ, Gauthier M, Pin JP, Dissous C.
A new family of receptor tyrosine kinases with a venus flytrap binding
domain in insects and other invertebrates activated by aminoacids.
PloS One, 2009, 4:e565.
Baeza-Garcia A, Pierce RJ, Gourbal B, Werkmeister E, Colinet D,
Reichhart JM, Dissous C, Coustau C.
Involvement of the cytokine MIF in the snail host immune response to the
parasite Schistosoma mansoni.
PLOS pathogens, 2010, in press.
Bachega JF, Navarro MV, Bleicher L, Bortoleto-Bugs RK, Dive D,
Hoffmann P, Viscogliosi E, Garratt RC.
Systematic structural studies of iron superoxide dismutases from human
parasites and a statistical coupling analysis of metal binding specificity.
Proteins. 2009, 77:26-37.
20
Contents
Beckmann S, Buro C, Dissous C Hirzmann J, Grevelding CG.
The Syk kinase SmTK4 of Schistosoma mansoni is involved in the
regulation of spermatogenesis and oogenesis.
PloS Pathogen, 2010, 6:e1000769.
Beckmann S, Quack T, Burmeister C, Buro C, Long T, Dissous C,
Grevelding CG.
Schistosoma mansoni : Signal transduction processes during the
development of the reproductive organs.
Parasitology, 2010, 137:497-520.
Daher W, Pierrot C, Kalamou H, Pinder JC, Margos G, Dive D, FrankeFayard B, Janse CJ, Khalife J.
Plasmodium falciparum dynein light chain 1 interacts with actin/myosin
during blood stage development.
J Biol Chem. 2010, 285:20180-20191.
Dissous C, Grevelding CG.
Piggy-Backing the concept of cancer drugs for schistosomiasis treatment
and beyond, a tangible perspective ?
Trends in Parasitology, 2010, in press
Long T, Cailliau K, Beckmann S, Browaeys E, Trolet J, Grevelding CG,
Dissous C.
Schistosoma mansoni polo-like kinase 1 : A mitotic kinase with key
functions in parasite reproduction.
Int J Parasitol, 2010, 40:1075-1086.
PhD
Arnaud AHIER
Directeur de thèse : Dissous C.
Etude des Recepteurs Tyrosine Kinase du parasite helminthe Schistosoma
mansoni- Decouverte des Recepteurs Venus Kinase, une nouvelle famille
de RTK.
Doctorat avec option éventuelle : Discipline Parasitologie-Biologie
Moleculaire
Université de Lille 2
12 décembre 2008
Aurore ACROUTE DIT VAMPOUILLE
Directeur de thèse : Khalife J. /Pierrot C.
Analyse des réponses cellulaires et moléculaires dans le paludisme
expérimental chez le rat : vers l’identification des mécanismes impliqués
dans la susceptibilité dépendante de l’âge.
Doctorat avec option éventuelle : Discipline Maladies infectieuses
15 décembre 2009
Florence DUBOIS-ABDESSELEM
Directeur de thèse : Pierce RJ.
Etude des histones déacetylases (HDACs) de classes I et III du parasite
plathelminthe Schistosoma mansoni.
Doctorat avec option éventuelle : Discipline Parasitologie-Biologie
Moleculaire
17 décembre 2009
Thavy LONG
Directeur de thèse : Dissous C.
Les Polo-like kinases de Shistosoma mansoni (SmPlK1 et SmSaK) :
caractérisation fonctionnelle et étude de leur régulation dans le processus
de mitose.
3 juin 2010
21
Contents
chromosomal genes that, together with the two Y. pestisspecific plasmids pMT1 [102 kb] and [9.6 kb] pPCP1, represent
the only genetic material acquired by Y. pestis since its
divergence from Y. pseudotuberculosis. In contrast, 149
pseudogenes and 317 genes absent from Y. pestis were
detected, indicating that as many as 13% of
Y. pseudotuberculosis genes no longer function in Y. pestis.
Extensive insertion sequence (IS)-mediated genome
rearrangements and reductive evolution through massive gene
loss (resulting in elimination and modification of pre-existing
gene expression pathways) appear to have been more
important than gene acquisition in the evolution of Y. pestis.
Y. pestis possesses 32 specific chromosomal genes, but it is
unlikely that their presence alone can explain the radically
different clinical pictures induced by this infectious agent when
compared with Y. pseudotuberculosis. Differential transcription
of species-conserved genes (75% of shared chromosomal
genes are 97% identical at the nucleotide level) may be at the
origin of this clinical disparity. Therefore, in a collaborative study
with the Institut Pasteur's Yersinia Laboratory and Genopole,
we compared overall gene expression patterns in
Y. pseudotuberculosis and Y. pestis in order to identify genes that
are differently transcribed in one species or the other under in
vitro culture conditions that match those encountered in vivo
as closely as possible. The DNA macroarrays included 4,204 PCR
fragments corresponding to 3,507 genes shared by both
species (i.e. > 90% nucleotide identity) and 697 genes differing
between the two species (i.e. presence/absence or presence of
an IS). Whole-transcriptome analysis revealed that the three
shared virulence factors - (i) the Ycs-Yop type III secretion system
(encoded by the 70-kb virulence plasmid pYV/pCD1 that is
responsible for injecting a number of cytotoxins and effectors
into host cells, circumventing the innate immunity processes
into host cells), (ii) the yersiniabactin siderophore and (iii) the
RovA transcriptional activator - are differentially transcribed in
the two pathogens. Genes encoding the first two were more
strongly expressed in Y. pestis and conversely, rovA was much
more transcribed in Y. pseudotuberculosis. Transcriptional
differences between the two species also suggested that a
number of genes encoding putative virulence factors (such as
the two Ail adhesins, Pla2 omptin and KatY catalase) play a
species-specific role.
Plague and Yersinia pestis
Florent SEBBANE, CR1 Inserm
Group Members :
Courcol René, PU-PH1 Univ Lille 2/CHRU
Simonet Michel, PU-PH1 Univ Lille 2/CHRU
Lemaître Nadine, MCU PH1 Univ Lille 2/CHRU
Marceau Michaël, MCU Univ Lille 2
Pradel Elizabeth, CR1 Inserm
Reboul Angeline, Technician Inserm
Lapiere Sarah, Technician Univ Lille 2
Ricard Isabelle, Technician IPL
Pierre François, PhD Student MENRT
Biki-Triki Sabrina, Master 2 Student
Key words :
Plague. Yersinia. Rodent infection model. Flea infection model.
Molecular microbiology. Virulence.
A - Research Report
The PYP team has emerged from Inserm U801. The group's
overall research theme is the study of the pathogenic properties
of a medically important bacterium that infects the human
intestine : Yersinia pseudotuberculosis. It typically causes ileitis
and mesenteric lymphadenitis, but it can sometimes provoke
immunopathological complications, such as reactive arthritis,
erythema nodosum or Kawasaki syndrome. The bacterium
usually remains in the digestive tract, but it can disseminate into
the body in immunocompromised hosts. Notably, this microorganism is the progenitor of Yersinia pestis, the causative agent
of plague. The goal of our research over the past 4 years has
been to study the genetics of bacterial virulence and, although
most of the work has been done so far on Y. pseudotuberculosis,
investigation of Y. pestis pathogenesis has been developed
recently and will be the main object of the future research of
the team.
1. Y. pseudotuberculosis genomics
and postgenomics
2. A novel pathogenicity island
in enteropathogenic Yersinia
Y. pseudotuberculosis is genetically closely related to Y. pestis,
and there is strong molecular evidence to suggest that
Y. pseudotuberculosis is the recent ancestor of the plague
bacillus (with divergence between 1,500 and 20,000 years ago).
However, despite their close genetic relationship, the two
micro-organisms differ radically in regard of their pathogenicity
(self-limiting enteritis for Y. pseudotuberculosis versus a
frequently disseminated, fatal infection for Y. pestis). Hence, we
reasoned that a comparative genomics analysis might enable
us to identify elements whose presence or absence is
responsible for the difference in bacterial pathogenicity.
Therefore, we have participated (as part of an international
academic consortium) in the annotation of the full
Y. pseudotuberculosis (strain IP32953) genome sequence (~4.8
Mb) and comparison with available Y. pestis sequences. The
analysis of identified differences across a panel of Yersinia
isolates from around the world revealed 32 Y. pestis
Bacterial genomic islands are genetic elements that are
horizontally transferred between distantly-related taxons. The
islands' foreign origin is supported by (i) codon usage and
guanine (G) & cytosine (C) content which often differ from the
host's chromosomal core and (ii) the presence of a range of
mobility genes involved in DNA transmission. Pathogenicity
islands (PAIs) are genomic islands harbouring virulence genes,
and their spread throughout the bacterial world promotes the
emergence of new pathogenic species. Although a few PAIs
have been described in distant genera, coherent evolutionary
scenarios describing successive transfers had not previously
been demonstrated. We discovered that Y. pseudotuberculosis
produces type IV pili, which contribute to bacterial
pathogenicity. These fimbriae are encoded by an eleven-gene
pil polycistronic unit : it exhibits a higher GC content than that
of the Yersinia chromosome and is present in only 40% of the
22
Contents
species' strains. After sequencing its flanking regions, we
established that the pil locus was located on a novel PAI that we
refer to as YAPI (for "Yersinia Adhesion Pathogenicity Island").
Despite its large size (98 kilobases), YAPI does not contain any
other virulence genes. Closely-related PAIs were detected in two
other enteric pathogens (Y. enterocolitica (YAPIent) and
Salmonella enterica (SPI-7)) and in the entomopathogenic
bacterium Photorhabdus luminescens (1 W14). Genomics
methods initially developed at the gene and/or nucleotide level
(i.e. comparisons of concatenated sequences, orthologue
frequency, gene order or dinucleotide usage) were combined
and applied to the homologous PAIs. The ancestral YAPI
emerged from a Salmonella genetic element and was then
acquired by Y. pseudotuberculosis and Y. enterocolitica prior to
speciation. In each bacterium, various DNA rearrangements
(e.g. gene deletions and additions) resulted in a segment, which
is common to the four islands (i.e. YAPIpst, YAPIent, SPI-7 and 1
W14) and others, which are specific to each bacterium. YAPI is
absent from the Y. pestis genome (the latter having recently
emerged from Y. pseudotuberculosis). Since YAPI can
spontaneously excise from Y. pseudotuberculosis, Y. pestis derives
from a YAPI-deleted clone.
Like the pil gene cluster, the superantigen-encoding gene ypm
has been laterally acquired by Y. pseudotuberculosis - most
probably by transduction after divergence from the Yersinia
progenitor. PCR assays on 270 unrelated strains from various
environmental and animal sources revealed a significant
association between ypm and pil in isolates. Fimbriae (and
especially type IV pili) may serve as bacteriophage receptors
at the bacterial cell surface. Hence, an evolutionary scenario
for the emergence of ypm-positive strains (reminiscent of that
proposed for the origin of enterotoxinogenic Vibrio cholerae)
would be the following : first, Yersinia would have acquired the
pil operon (the counterpart in V. cholerae is the PAI[VIP]-borne
tcp operon) via YAPI transfer before it speciated and second,
some pil-positive strains would have been infected and
lysogenized by a ypm-encoding prophage (the counterpart in
V. cholerae is a filamentous temperate phage CTXΦ encoding
cholera toxin) using type IV pili as receptors.
regulators of complete systems via reverse genetics, we
sought to determine the ability of the resulting 24 mutants to
survive in vitro when confronted with the various types of
stresses encountered by the bacterium in the digestive tract.
Ten mutants showed levels of resistance to low pH and/or
high osmotic pressure, oxidative stress, the presence of bile or
antimicrobial peptides, which differed from those of the wild
type. However, only 4 (phoP, rstA-like, yfhA-like and above all
ompR) were less virulent than the wild type when
administered orally to mice. Bearing in mind the limitations of
our experimental models, a small set of 2CSs thus appears to
influence the virulence of Y. pseudotuberculosis. Although
some mutant phenotypes were consistent with those (when
known) of the corresponding, putative orthologue mutants in
other pathogenic species, several response regulators
behaved differently in Y. pseudotuberculosis ; these include the
PmrA, PhoP, and ArcA-like response regulators, which were
found to control bile salt resistance in a different manner to
that observed in Salmonella.
A seven-gene polycistronic unit (the pmrF operon) is essential
for the biogenesis, export and the addition of 4-deoxy-4
amino-L arabinose (hereafter referred to as 4-aminoarabinose)
to the lipid moiety of Y. pseudotuberculosis lipo-polysaccharide
(LPS), as we demonstrated by MALDI-TOF mass spectrometry
of lipid A from wild-type and pmrF-mutated strains. This
modification reduces the net negative charge of LPS and, as a
consequence, minimises initial electrostatic interactions
between the bacterium and cationic antimicrobial peptides
such as polymyxin B. Therefore, the addition of
4-aminoarabinose to LPS contributes to bacterial resistance
to these compounds. PhoP, the response regulator of a 2CS
(PhoP/PhoQ, mediating bacterial adaptation to Mg2+- and
Ca2+-limiting environments) activates the 4-aminoarabinose
substitution of lipid A by interacting directly with the pmrF
operon promoter region. The Salmonella chromosome
harbours a homologous pmrF operon, which is activated by
PhoP and PmrA (a regulator responding to high Fe3+ levels
detected by the 2CS PmrB sensor). In contrast, we found
that Y. pseudotuberculosis pmrF upregulation is PmrAindependent. We also established that pmrF operon
transcription in Y. pseudotuberculosis is regulated by a LysRtype activator, whose expression is induced by iron starvation.
The latter also mediates bacterial virulence in vivo, because its
inactivation is associated with reduced Yersinia pathogenicity
in the orally infected mouse model.
Taken as a whole, our data revealed that transcriptional
network remodelling may (along with genome evolution) be
a major cause of phenotypic adaptation and thus species
divergence in Y. pseudotuberculosis.
3. Two component-systems
and Y. pseudotuberculosis virulence
In bacteria, very rapid adaptation to a changing (and generally
hostile) environment is essential for survival. To this end, a
number of pathways enable bacteria to interact with external
signals. One of the most effective pathways involves twocomponent systems (2CSs). The prototype is constituted by a
membrane-located histidine kinase (sensing changes in the
environment) coupled to a cytoplasmic response regulator
(either an activator or a repressor) capable of acting on the
transcription of a set of target genes referred to as a regulon.
Once the stimulus has been sensed, the sensor
phosphorylates itself and then transfers the phosphate group
to the cognate response regulator. This reaction induces a
cellular response : the two bacterial proteins thus create a
circuit, which converts an external danger signal into an
appropriate gene transcription response.
An in silico analysis of the Y. pseudotuberculosis genome has
revealed the presence of 24 2CSs, together with one orphan
sensor (i.e. lacking a regulator partner) and three orphan
regulators. Following inactivation of each of the response
4. Y. pestis and host interactions in vivo
Plague is a zoonosis caused by Y. pestis. Bubonic plague, the
most common form of the disease in rodents and humans, is
acquired from the bite of an infected flea. It is characterized
by a painful, swollen lymph node termed a bubo. In the
absence of early antibiotic treatment, bubonic plague usually
progresses to bacteremia, systemic spreading and lifethreatening Gram-negative sepsis.
We developed a model of bubonic plague in the rat -a major
natural host of Y. pestis- to understand the cellular and
molecular physiopathology of the illness. After intra-dermal
inoculation of Y. pestis, disease progression and histopathology
23
Contents
in this rodent species closely resemble those seen in humans.
In vivo microarray analysis of Y. pestis gene expression revealed
an adaptive response to certain toxic effectors discharged by
polymorphonuclear neutrophils (PMNs) in infected tissues.
PMNs recruited to the infected lymph node expressed
abundant inducible NO synthase, and several Y. pestis homologs
of genes involved in the protective response to reactive
nitrogen species were up-regulated in the bubo. Mutation of
one of these genes, which codes for the Hmp flavohemoglobin
that detoxifies NO, attenuated bacterial virulence.
The yopJ gene encodes a toxin which is secreted by Y. pestis
into phagocytes, inducing apoptosis and inhibiting the
release of TNF in vitro. We evaluated its role in the abovementioned animal model and found that deletion of yopJ
resulted in a twofold reduction in the number of apoptotic
immune cells in the bubo and a threefold increase in serum
TNF levels. Despite its anti-immunity activity, YopJ was not
required for full virulence.
We also set up an arthropod-borne transmission model of
plague in mouse and we demonstrated that after flea bites
Y. pestis can disseminate in rodents by two distinct routes : one
leading to bubo formation and dependent upon a bacterial
plasminogen activator (Pla) and another resulting in primary
septicaemic plague without buboes. By contrast, the artificial
transmission route of Y. pestis (i.e. intra-dermal inoculation
through a needle) results only in bubonic plague. Using our
flea-borne transmission model, we showed that the
recombinant fusion protein vaccine consisting of the Y. pestis
F1 and V antigens protects against natural infestation by the
plague bacillus.
Publications
2007
Chauvaux S, Rosso ML, Frangeul L, Lacroix C, Labarre L, Schiavo A,
Marceau M, Dillies MA, Foulon J, Coppée JY, Médigue C, Simonet M,
Carniel E.
Transcriptome analysis of Yersinia pestis in human plasma : an approach
for discovering genes involved in septicaemic plague,
Microbiology, 2007, 153:3112-3123.
Foligne B, Dessein R, Marceau M, Poiret S, Chamaillard M, Pot B,
Simonet M, Daniel C.
Prevention and treatment of colitis with Lactococcus lactis secreting the
immunomodulatory Yersinia LcrV protein.
Gastroenterology, 2007, 133:862-874.
Nehme NT, Liegeois S, Kele B, Giammarinaro P, Pradel E,
Hoffmann JA, Ewbank JJ, Ferrandon D.
A model of bacterial intestinal infections in Drosophila melanogaster.
PLoS Pathogens, 2007, 3: e173.
Pradel E, Zhang Y, Pujol N, Matsuyama T, Bargmann CI, Ewbank JJ.
Detection and avoidance of a natural product from the pathogenic
bacterium Serratia marcescens by Caenorhabditis elegans.
Proceed Nat Acad Sci USA , 2007, 104:2295-2300.
Sirard JC, Vignal C, Dessein R, Chamaillard M.
Nod-like receptors: cytosolic watchdogs for immunity against pathogens.
PLoS. Pathogens, 2007, 3: e152.
Vadyvaloo V, Jarrett CO, Sturdevant D, Sebbane F, Hinnebusch BJ.
Analysis of Yersinia pestis gene expression in the flea vector.
Adv Exper Med Biol, 2007, 603:192-200.
Vincent P, Salo E, Skurnik M, Fukushima H, Simonet M.
Similarities of Kawasaki disease and Yersinia pseudotuberculosis infection
epidemiology.
Pediat Infect Dis J, 2007, 26:629-631.
development 2010
2008
Flamez C, Ricard I, Arafah S, Simonet M, Marceau M.
Phenotypic analysis of Yersinia pseudotuberculosis response regulator
mutants : new insights into two-component system regular plasticity in
bacteria.
Int J Med Microbiol, 2008, 298:193-203.
The overall objective of our research is geared towards a
better understanding of arthropod-borne diseases in general
and plague in particular. Plague is a flea-borne re-emerging
zoonosis caused by the bacterium Yersinia pestis. Plague is still
a worldwide public health issue accentuated by the
emergence of multiply-antibiotic resistant strains of Y. pestis
and the potential criminal release of this highly virulent
bacterium. Our recent investigation of how Y. pestis adapts to
the immune response and other host factors in the infected
lymph node has revealed that Y. pestis has to resist to
antimicrobial molecules that are secreted by PMNs in infected
tissues in addition to inhibit phagocytosis. Our project aims
at 1) improving existing therapies and developing new
therapeutics against plague and 2) identifying new molecular
mechanisms used by Y. pestis to evade immune responses.
Vincent P, Leclercq A, Martin L, The Yersinia Surveillance
Network, Duez JM, Simonet M, Carniel E.
Sudden onset pf pseudotuberculosis in humans.
Emerg Infect Dis, 2008, 14:1119-1122.
2009
Arafah S, Rosso ML, Rehaume L, Hancock RE, Simonet M, Marceau M.
An iron-regulated LysR-type element mediates antimicrobial peptide
resistance and virulence in Yersinia pseudotuberculosis.
Microbiology, 2009, 155:2168-2181.
Daniel C, Sebbane F, Poiret S, Goudercourt D, Dewulf J, Mullet C,
Simonet M, Pot B
Protection against Yersinia pseudotuberculosis infection conferred by a
Lactococcus lactis mucosal delivery vector secreting LcrV.
Vaccine, 2009, 27:1141-1144.
Sebbane F, Jarrett C, Gardner D, Long D, Hinnebusch BJ.
The Yersinia pestis caf1M1A1 fimbrial capsule operon promotes
transmission by flea bite in a mouse model of bubonic plague.
Infect Immun. 2009, 77:1222-1129.
24
Contents
2010
Ayyadurai S, Sebbane F, Raoult D, Drancourt M.
Body lice, Yersinia pestis orientalis, and black death.
Emerg Infect Dis, 2010, 16:892-893.
Loïez C, Carnoy C, Decoene C, Pradel E, Fichel C, Courcol R, Wallet F.
First case of postaneurysmal prosthetic vascular infection due to a
nonsuperantigenic Yersinia pseudotuberculosis strain.
J Clin Microbiol, 2010,48:3024-3026.
Moreau K, Lacas-Gervais S, Fujita N, Sebbane F, Yoshimori T,
Simonet M, Lafont F.
Autophagosome can support Yersinia pseudotuberculosis replication in
macrophages.
Cell Microbiology, 2010, 12:1108-1123
Vadyvaloo V, Jarrett C, Sturdevant DE, Sebbane F, Hinnebusch BJ.
Transit through the Flea Vector Induces a pretransmission innate immunity
resistance phenotype in Yersinia pestis.
PLoS Pathogens, 2010, 6:e1000783.
Van Maele L, Carnoy C, Cayet D, Songhet P, Dumoutier L, Ferrero I,
Janot L, Erard F, Bertout J, Leger H, Sebbane F, Benecke A,
Renauld JC, Hardt WD, Ryffel B, Sirard JC.
TLR5 signaling stimulates the innate production of IL-17 and IL-22 by
CD3negCD127+ immune cells in spleen and mucosa.
J Immunol, 2010, 185:1177-1185.
PhD
Claire FLAMEZ
Directeur de thèse : Simonet Michel
« Systèmes à deux composants et virulence de Yersinia pseudotuberculosis »
2007.
Sonia ARAFAH
« Induction par un stress de la résistance aux peptides anti-microbiens
chez Yersini » pseudotuberculosis,
2008.
Rodrigue DESSEIN
« Réponse immunitaire de la muqueuse intestinale à un agression par
Yersinia pseudotuberculosis »
2008.
HDR
Florent SEBBANE
2008
25
Contents
research represents a continuum from very basic aspects all
the way to applications and clinical investigations, with the
help of collaborations with other French (within and outside
of the Center for Infection and Immunity of Lille) and foreign
laboratories, including collaborators in the framework of the
programmes of the European Union, in which we participate
very actively.
Bacterial Respiratory
Infections : Pertussis
and Tuberculosis
Camille LOCHT, DRE Inserm
The choice of the two bacterial models is motivated by the
fact that both infect the respiratory tract of man, but one is
very pathogenic (virtually every immunologically naive
individual infected with B. pertussis will develop pertussis),
while the other is very poorly pathogenic (only approximately
5 to 10 % of the M. tuberculosis-infected individuals will
develop active TB during their lifetime). On the other hand,
M. tuberculosis is much more successful than B. pertussis, as
approximately 1/3 of the world population is infected with the
tubercle bacillus. B. pertussis is essentially, but not exclusively,
an extracellular pathogen, while M. tuberculosis is essentially,
but not exclusively, an intracellular pathogen. For both
infections vaccines are available, but the re-emergence of
both diseases indicates that these vaccines will have to be
improved. Nevertheless, the availability of protective vaccines
provides us with tools that will allow us to study the
mechanisms of protective immunity.
Since the pathogenic potential of a virulent bacterium largely
depends on its interaction with the host, it involves essentially
factors that are either secreted by the pathogen or that are
displayed at its surface. The study of the bacterial envelope
constitutes therefore our first research theme. Many genes
involved in virulence are regulated in response to
environmental conditions. The study on the regulation of the
production of the virulence factors constitutes thus our
second research theme. Finally, the third theme concerns
more specifically research applied to the development of new
vaccines. In that context we wish to fully integrate all the
knowledge obtained in themes 1 and 2 to develop novel
vaccine approaches against pertussis and TB.
Group Members :
Antoine Rudy, CR1 Inserm
Baulard Alain, DR2 Inserm
Coutte Loïc, CR2 Inserm
Jacobs-Dubuisson Françoise, DR2 CNRS
Mielcarek Nathalie, CR1 Inserm
Supply Philip, DR2 CNRS
Verwaerde Claudie, CR2 IPL
Vidal Carine, CR1 IPL
Debrie Anne-Sophie, CE1 IPL
Raze Dominique, IR1 Inserm
Willery Eve, CE1 IPL
Delhoute Aline, Technician Inserm
Lecher Sophie, Technician IPL
Loyens Marc, Technician IPL
Baux Catherine, Postdoctoral fellow IPL
Feunou-Feunou Pascal, Postdoctoral fellow Inserm
Guillet Nathalie, Postdoctoral fellow IPL
Herrou Julien, Postdoctoral fellow IPL
Mille Céline, Postdoctoral fellow IPL
Roux Xavier, Postdoctoral fellow Inserm
Segers Jérôme, Postdoctoral fellow IPL
Blondiau Nicolas, PhD Student Univ Lille 2
Chaari Hana, PhD Student IPL/région
Delattre Anne-Sophie, PhD Student IPL/région
Dupre Elian, PhD Student MENRT
Lebrun Pierre, PhD Student Univ Lille 2
Key words :
Vaccins. B. pertussis. M. tuberculosis. Adhesins. Toxins. Genomic
typing. Gene regulation. Virulence mechanisms. Diagnostics.
Anti-microbials.
1. The bacterial envelope
The envelope contains molecular structures that interact
directly with host cells, and that allow the bacteria to adhere
or to modulate the physiology of the host. In addition, the
envelope permits the importation of molecules required for
bacterial growth and the export of substances that may act at
a distance within the host, such as bacterial toxins.
A - Research Report
The “Bacterial Respiratory Infections : Pertussis and
Tuberculosis” (BRIPT) team is one of the teams of the recently
created Center for Infection and Immunity of Lille (CIIL, Inserm
U1019, CNRS UMR8204). Its research activity is focused on the
molecular pathogenesis of bacterial respiratory infections,
especially pertussis and tuberculosis. Today, respiratory
infections, many of which are caused by pathogenic bacteria,
are still among the world’s most successful killers. Tuberculosis
(TB) is the cause of approximately 2 million annual deaths.
Other respiratory diseases, such as pertussis, the existence of
which had almost been forgotten, show a dramatic reemergence at a global level, including in European countries
with wide vaccine coverage. The objectives of the team is (i)
to study the molecular details of the pathogenic mechanisms
of Bordetella pertussis and Mycobactrium tuberculosis and (ii)
to try to use this knowledge to develop novel approaches to
design better vaccines, new therapeutic molecules and new
diagnostic or molecular typing methods, in other words, to
help solve unsolved problems with pertussis and TB. Our
1. 1. Bordetella pertussis
1. 1. 1. Filamentous haemagglutinin. We have extensively
studied the secretion mechanism of filamentous
haemagglutinin (FHA), the major B. pertussis adhesin. It is a
220-kDa protein that is matured from a 367-kDa precursor. We
have shown that FHA is secreted by a mechanism we have
named “Two-partner secretion system” (TPS) that requires an
accessory outer-membrane protein (in this case FhaC) and an
N-proximal, conserved “TPS” secretion signal. In collaboration
with V. Villeret (CNRS/Institut Pasteur de Lille) we have
determined the crystal structure of the FHA TPS domain and
of FhaC. FhaC forms a transmembrane 16-stranded ß barrel
preceded by two periplasmic POTRA domains that recognize
FHA in a non-native conformation. We have studied the
interaction between the two molecules by various biophysical
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protects this domain against proteolytic degradation and is
also crucial for the antigenicity of the protein (see below). This
methylation is catalysed by specific mycobacterial HBHAmethyltransferases.
Genes encoding HBHA are present in many mycobacteria,
both pathogenic and non-pathogenic. We found that
M. smegmatis produces a HBHA-like protein lacking adhesive
properties, indicating that the adhesin in pathogenic
mycobacteria evolved from non-adhesin molecules in nonpathogenic mycobacteria.
and biochemical methods and have deciphered the nature of
the secretion signal by introducing point mutations. Upon
secretion, FHA is processed by the protease SphB1, an
autotransporter with a lipid moiety anchored in the outer
membrane. The structure of SphB1 has been studied using
biophysical techniques, including dynamic light scattering,
cross-linking and electrophysiology. We found that the
passenger domain enhances the auto-association of SphB1,
and thus possibly facilitates auto-maturation in vivo, and that
it also stabilizes the pore after secretion. Studies of the sorting
of SphB1 have shown the importance of residues +2 to +4 for
its addressing to the outer membrane. Finally, we have
identified two periplasmic chaperones that interact with FHA.
One is a peptidyl-prolyl isomerase, Par27, and the other a
protease, DegP. We have characterized the structure and
function of Par27.
1. 1. 2. The Bordetella Bug family. We have identified novel
periplasmic proteins involved in the uptake of small
molecules, named Bug (for Bordetella uptake genes). They are
part of a TTT system (for tripartite tricarboxylic transport) that
is composed of two membrane proteins in addition to the
Bug. We have studied in detail one of them, involved in citrate
uptake. Citrate binds to the cognate Bug protein and is then
transported through the inner membrane via the two
membrane proteins. In parallel, the citrate-bound Bug
interacts with the sensor protein of a two-component system,
which triggers the expression of the citrate transport system.
In collaboration with V. Villeret, we have determined the
crystal structure of several Bugs, which feature a Venus flytrap
fold with precise arrangements of hydrogen bonds for their
specific ligand binding.
We have also studied DctP6 and DctP7 and have obtained the
crystal structures of the two proteins (coll. V. Villeret). DctP6
and 7 have typical Venus fly trap structures like Bugs, but their
mode of ligand binding is different. They are prototypes of a
new subfamily of PBPs specific for pyroglutamate.
2. Genomics and gene regulation
Since the entire genomes of the two model micro-organisms
have been sequenced, we study genetic regulation at a global
regulome level. In addition, certain polymorphic sequences
identified in the genomes have enabled us to develop new
tools for diagnostics and molecular typing.
1. 2. Mycobacterium tuberculosis
The work on FHA has inspired us to study adhesins of
M. tuberculosis, and has led to the discovery of heparinbinding haemagglutinin (HBHA), responsible for binding to
non-phagocytic cells and for extrapulmonary dissemination.
We have characterised the interaction forces between HBHA
and its receptors (sulphated glycoconjugates) and mapped
the distribution of the adhesin on the surface of the
mycobacteria by atomic force microscopy. We have used in
silico models of the secondary or tertiary structures, combined
with with SAXS (Small-angle X-ray scattering) and CD (circular
dichroism) to obtain structural elements. These studies
indicate that the N-terminal domain is rich in α helices,
whereas the C-terminal, lysine-rich region is relatively
unstructured. A coiled-coil region in the N-terminal domain,
shares sequence similarities with actin-binding proteins. In
vitro evidence that specific interaction forces exists between
HBHA and actin have been demonstrated using AFM (Atomic
Force Microscopy) in collaboration with Yves Dufrene (UCL,
Belgium). In addition, this domain is responsible for
homotypic interactions between HBHA molecules leading to
the formation of multimers.
We have shown that the lysine-rich domain, which is
important for the interaction of HBHA with its receptors,
shows a complex methylation pattern. The methylation
The production of most B. pertussis virulence factors is
regulated by the two-component system BvgA/S. BvgS is the
sensor protein, and BvgA is the activator that induces the
expression of the virulence genes, named vag (for “ virulence
activated genes ”) and represses the expression of the vrg (for
“virulence repressed genes”). We have developed tools to
study the regulome of B. pertussis, including transcriptomics,
proteomics and functional genomics. They have allowed us to
identify the Bug proteins (see above), HotA/B, a homolog of
pertussis toxin (PTX), as well as other vags and vrgs. We have
also inactivated all two-component systems of B. pertussis, as
well as the non-essential ECF-type sigma factors. These studies
have established a link between the Bvg regulon and the
respiratory chain.
BvgS is composed of two periplasmic PBP-like domains,
followed by a transmembrane helix, a PAS domain and several
modules involved in phosphate transfer to BvgA. We have
solved the crystal structure of the PBP-like domain 2. It adopts
a typical PBP fold. By fluorescence spectroscopy, we have
shown that it binds some amino acids found in the growth
medium for B. pertussis. The PBP1 domain was not crystallized,
but its structure was modelled based on the PBP2 domain. By
site-directed mutagenesis, we have modified residues
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with one of them allowed us to reduce the dosage of ETH threefold. Treatment of M. tuberculosis with this compound also
resulted in an increased sensitivity to TAZ, but not to ISO. Since
this family of prodrugs is also used against M. leprae, the use of
EthR inhibitors may also be beneficial for treatment of leprosy.
potentially involved in ligand binding in both domains. Some
of these variants are insensitive to modulators. We have also
generated BvgS hybrids containing combinations of BvgS
domains from various strains that display a wide range of
sensitivities to modulators, and we have thereby identified a
highly sensitive variant. The mutation in the histidine kinase
domain seems to influence the modulator effect on the
phosphorylation reaction. By exchanging PBP domains, we
have shown that only the PBP2 domain from B. bronchiseptica
is capable of conferring an enhanced sensitivity to the
virulence modulators.
2. 2. 1. Mycobacterial Interspersed Repetitive Units. The
study of mycobacterial two-component systems has led to the
discovery of novel genetic elements that we have named
MIRU (for Mycobacterial Interspersed Repetitive Units). They
are small genetic elements coding for small peptides and are
generally localised within operons to regulate the expression
levels of downstream cistrons. The polymorphism displayed
by some of them has allowed us to develop a powerful new
typing tool for the M. tuberculosis complex. The MIRU typing
method appears now to be the method of choice for many
laboratories, including reference laboratories such as the CDC.
It is fast and provides the results in a numeric format perfectly
portable between laboratories. Using this tool, we have shown
the clonality of M. tuberculosis. We have now adapted an
optimised set of MIRU loci, which has resulted in a highly
discriminatory method, and developed a freely accessible
multifunctional MIRU-VNTRplus database (www.miruvntrplus.org). We have also adapted the system for the typing
of Mycobacterium paratuberculosis and Mycobacterium
ulcerans and have used this tool for phylogenetic analyses.
This has led to a new hypothesis on the origin of tuberculosis
and the discovery of Mycobacterium prototuberculosis. We also
found that the complex comprises two independent clades,
one including exclusively human pathogens and the other
including human and animal isolates. By using Bayesian
statistics and data on the variability of MIRU-VNTR markers,
we could estimate the age of the complex to about 40,000
years, approximately coinciding with modern human
expansion out of Africa.
3. Antigen presentation
and vaccine development
Vaccination against whooping cough, initially with firstgeneration vaccines, composed of whole, inactivated bacteria,
and now more and more with second-generation, acellular
vaccines, containing essentially PTX and FHA, has been very
effective in reducing the disease in the classical age group of 6
months to 10 years. However, infant pertussis is still very
common, and this problem cannot be solved with the current
vaccines. We have shown that, unexpectedly in view of the
neonatal immaturity of the immune system, very young infants
(median of 2 months) infected with B. pertussis are able to
mount a potent cellular immune response to B. pertussis
antigens. This response is not obtained when the children were
vaccinated with acellular vaccines. Based on these observations,
we have developed a live attenuated vaccine candidate to be
administered by the nasal route. The vaccine strain, named
BPZE1, was constructed by modifying the ptx gene so that PTX
remains immunogenic but is non-toxic. Furthermore, the gene
encoding the dermonecrotic toxin was removed, and the ampG
gene of B. pertussis was replaced by that of E. coli, resulting in
the abolition of tracheal cytotoxin production. BPZE1 colonises
the mouse respiratory tract very well, but it causes no
detectable lung inflammation. Nevertheless, it protects mice,
including infant mice (3 wk-old), after a single intranasal
administration, unlike the acellular vaccine. We dissected the
host immune functions necessary for protection using passive
transfer to SCID mice and found that it is mediated by both
antibodies and CD4+ Th1, but not by CD8+ T cells. In addition,
BPZE1 immunization protected against B. parapertussis
infection, but this protection could only be transferred by T cells.
We assessed the genetic stability of BPZE1 after 20 and 27
weeks of continuous passaging in vitro and in vivo, respectively.
After these passages, 8 hemolytic colonies were analyzed by
PCR for the absence of dnt and B. pertussis ampG and the
presence of E. coli ampG, by DNA sequencing for the presence
of the two ptx point mutations and by DNA microarrays for
2. 2. 2. Regulation of pro-drug activation. Most current antiTB drugs are actually ”prodrugs” that must be metabolically
activated to manifest their toxicity. As such, the
monooxygenase EthA activates the second-line drug
ethionamide (ETH), thiacetazone (TAZ) and isoxyl (ISO).
Overproduction of EthA in recombinant mycobacteria
drastically increases sensitivity to the prodrugs. We found that
the production of EthA is under the control of the
transcriptional regulator EthR. In collaboration with V. Villeret,
we have determined the crystal structure of EthR and found a
ligand bound to EthR that may inhibit its repressor. This led to
a search for EthR inhibitors that can be used to increase the
sensitivity of M. tuberculosis to ETH. In collaboration with
B. Déprez (U761) we developed a drug design process through
the exploitation of EthR-compound interactions, which was
used to design a library of drug-like compounds that were
screened for their ability to interfere with the DNA-recognition
function of EthR. The drug-likeness of the two best compounds
was documented in vitro and in vivo. The combination of ETH
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we will continue to participate very actively. The activities of our
team remains structured in three major themes : (i) the bacterial
envelope, (ii) genetic regulation and genomics, and (iii) antigen
presentation and vaccine development.
global genomic stability. In addition, the protective capacity of
BPZE1 was evaluated after the passages. No genetic or
protective difference was detected between the passaged
bacteria and non-passaged BPZE1. Together these studies are
designed to be incorporated into a pre-clinical file that will be
useful for the development of clinical trials.
Publications
We have studied the immunological differences between
latently infected individuals and patients with active TB and
found that during latency infected subjects mount a very strong
T cell response to methylated, but not to non-methylated
HBHA, whereas this is not the case for patients with active TB.
The T cell response of latently infected subjects is characterised
by a strong IFN-γ production by both CD4+ and CD8+, as well as
by a strong cytotoxic and bactericidal activity of the CD8+ T cells,
indicating that during latency, infected subjects mount all
known protective parameters, probably preventing them from
progressing to active disease. The poor HBHA-specific T cell
responses of the TB patients could be related to the induction
of CD4+CD25+FOXP3+CD127- regulatory T cells that downmodulate the HBHA-specific T cell responses in these patients.
In vitro depletion of these cells results in an increase in HBHAspecific IFN-γ responses by the TB patients. Interestingly, these
Treg cells could be induced in vitro from the PBMC of latently
infected individuals, but not from non-infected controls, by
incubating them with BCG and TGF-ß, providing thereby a
mechanistic model for reactivation of TB.
These findings suggest that HBHA is a good protective antigen.
We have determined that HBHA provides protection in mice
and in the guinea pig model in the framework of the TB-VAC EU
project. We also investigated the boosting effect of HBHA on
the protective immunity induced by BCG and found that two
injections of HBHA+DDA-MPL 8 months after BCG priming
strongly increases the protection. Our study also highlighted
the importance of the time period between priming and
boosting on the protective efficacy. We also compared the
HBHA specific IFN-γ in these mice and observed a negative
correlation between the bacterial load and the amount of
IFN-γ produced upon stimulation with HBHA, suggesting
that the HBHA specific IFN-γ response constitutes a correlate
of protection.
2007
Alsteens D, Dague E, Rouxhet P, Baulard A, Dufrene Y.
Direct measurement of hydrophobic forces on cell surfaces using AFM.
Langmuir, 2007, 23:11977-11979.
Bayliss R, Harris R, Coutte L, Monier A, Fronzes R, Christie PJ,
Driscoll PC, Waksman,G.
NMR structure of a complex between the VirB9/VirB7 interaction domains
of the pKM101 type IV secretion system.
Proc Natl Acad Sci USA, 2007, 104:1673-1678.
Becq J, Gutierrez MC, Rosas-Magallanes V, Rauzier J, Gicquel B,
Neyrolles O, Deschavanne P.
Contribution of horizontally acquired genomic islands to the evolution
of the tubercle bacilli.
Mol Biol Evol, 207, 24:1861-1871.
Bethunaickan R, Baulard AR, Locht C, Raja A.
Antibody response in pulmonary tuberculosis against recombinant
27kDa (MPT51, Rv3803c) protein of Mycobacterium tuberculosis.
Scand J Infect Dis, 2007, 30:1-8.
Biet F, Angela de Melo Marques M, Grayon M, Xavier de Silveira EK,
Brennan PJ, Drobecq H, Raze D, Vidal Pessolani MC, Locht C,
Menozzi FD.
Mycobacterium smegmatis produces an HBHA homologue which is not
involved in epithelial adherence.
Clantin B, Delattre AS, Rucktooa P, Saint N, Méli A. Locht C, JacobDubuisson F, Villeret V.
Structure of the membrane protein FhaC : a member of Omp85- TpsB
transporter superfamily.
Science, 2007, 317:957- 961.
Dague E, Alsteens D, Latgé JP, Verbelen C, Raze D, Baulard AR,
Dufrene,Y.
Chemical force microscopy of single live cells.
Nano Lett, 2007, 10:3026-3030.
development 2010
Feunou P, Vanwetswinkel S, Gaudray F, Goldman M, Matthys P,
Braun MY.
FoxP3+CD25+ T regulatory cells stimulate IFN-gamma-independent
CD152-mediated activation of tryptophan catabolism that provides
dendritic cells with immune regulatory activity in mice unresponsive to
staphylococcal enterotoxin B.
J Immunol, 2007, 179:910-917.
The general objective of our team remains the same as in
previous years, i. e. to better understand the strategies adopted
by pathogenic bacteria of the respiratory tract and, as a
consequence, to obtain better tools for diagnostics, for the
design of more efficient anti-microbial compounds and for the
development of more appropriate vaccination strategies. We
will maintain our main focus on B. pertussis and M. tuberculosis
as models and will further strengthen the continuum from very
basic aspects all the way to applications and clinical
investigations, with the help of collaborations with other French
(within and outside of the Center for Infection and Immunity of
Lille) and foreign laboratories, including collaborators in the
framework of the programmes of the European Union, in which
Godreuil S, Torrea G, Terru D, Chevenet F, Diagbouga S, Supply P,
Van de Perre P, Carriere C, Banuls AL. 2007.
First molecular epidemiology study of Mycobacterium tuberculosis in
Burkina Faso.
J Clin Microbiol, 2007, 45:921-927.
Herrou J, Bompard C, Antoine R, Leroy A, Rucktooa P, Hot D,
Huvent I, Locht C, Villeret V, Jacob-Dubuisson F.
Structure–based mechanism of ligand binding for periplasmic solutebinding protein of the Bug family.
J Mol Biol, 2007, 373:954-964.
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Hodak H, Jacob-Dubuisson F.
Current challenges in autotransport and two-partner protein secretion
pathways.
Res Microbiol,2007, 8-9:631-637.
Segers J, Laumonier C, Burtea C, Laurent S, Elst LV, Müller, RN.
From phage display to magnetophage, a new tool for magnetic
resonance molecular imaging.
Bioconjug Chem, 2007, 18:1251-1258.
Host H, Drobecq H, Locht C, Menozzi F D.
Enzymatic methylation of the Mycobacterium tuberculosis heparinbinding haemagglutinin.
FEMS Microbiol Lett, 2007, 268:144-150.
Srivastava V, Rouanet C, Srivastava R, Ramalingam B, Locht C,
Srivastava BS.
Macrophage-specific Mycobacterium tuberculosis genes : identification
by green fluorescent protein and kanamycin resistance selection.
Microbiology, 2007, 153:659-566.
Hougardy JM, Place S, Hildebrand M, Drowart A, Debrie AS,
Locht C, Mascart F.
Regulatory T cells depress immune responses to protective antigens in
active tubeculosis.
Am J Respir Crit Care Med, 2007, 176:409-416.
Thibault VC, Grayon M, Boschiroli ML, Hubbans C, Overduin P,
Stevenson K, Gutierrez MC, Supply P, F. Biet F.
New variable−number tandem−repeat markers for typing
Mycobacterium avium subsp. Paratuberculosis and M. avium strains:
comparison with IS900 and IS1245 restriction fragment length
polymorphism typing.
J Clin Microbiol, 2007, 45:2404−2410.
Hougardy JM, Schepers K, Place S. Drowart A, Lechevin V,
Verscheure V, Debrie AS, Doherty TM, Van Vooren JP, Locht C,
Mascart F.
Heparin-binding hemagglutinin-induced IFN-γ release as a diagnostic
tool for latent tuberculosis.
PLoS One 2, 2007, e926.
Van Soolingen D, van Ingen J, Kremer K, Ferreira S, de Haas P,
Sebek M, Borgdorff M, Supply P.
VNTR typing as the next gold standard in the molecular epidemiology of
tuberculosis. 17th European Congress of Clinical Microbiology and
Infectious Diseases, and 25th International Congress of Chemotherapy.
Internat J Antimicrob Agents, 2007, 29 Suppl. 2:S17.
Hougardy JM, Verscheure V, Locht C, Mascart F.
In vivo expansino of CD4+CD25highFOXP3+CD127low/- regulatory T
cells from peripheral blood lymphocytes of healthy Mycobacterium
tuberculosis-infected humans.
Microb Infect, 2007, 9:1325-1332.
Verbelen C, Dupres V, Raze D, Dewitte F, Locht C, Dufrêne YF.
Single-molecule force spectroscopy of mycobacterial adhesin-adhesin
interactions.
J Bacteriol, 2007, 189:8801-8806.
Jauréguy F, Ioos V, Marzouk P, Hornstein M, Picard B, Gutierrez MC,
Valeyre D.
Mycobacterium heckeshornense : an emerging pathogen responsible for
a recurrent lung infection.
J Infect, 2007, 54:e33-35.
Yaradou DF, Raze D, Ginevra C, Ader F, Doleans-Jordheim A,
Vandenesch F, Menozzi FD, Etienne J, Jarraud S
Zinc-dependent cytoadherence of Legionella pneumophila to human
alveolarepithelial cells in vitro.
Microb Pathog, 2007, 43:234-242.
Ködmön C, Niemann S, Gutierrez MC. Sola C, Rastogi N, Lukacs J,
Somoskövi A.
Molecular clues of a microepidemy among homeless tuberculosis patients
in Budapest due to a new and local Mycobacterium tuberculosis clade.
Infect Genet Evol, 2007, 7:632-635.
2008
Abgrall S, Sereni D, Autran B, Carcelain G, Bourgarit A, Lagrange PH.
Anti-PGL-Tb1 responses as an indicator of the immune restoration
syndrome in HIV-TB patients.
Tuberculosis, 2008, 88, 453-461.
Locht C, Rouanet C, Hougardy JM, Mascart F.
How a different look at latency can help to develop novel diagnostics and
vaccines aginst tuberculosis.
Expert Opin Biol Ther, 2007, 7:1665-1677.
Ader F, Le Berre R, Fackeure R, Raze D, Menozzi FD, Viget N, Faure K,
Kipnis E, Guery B, Jarraud S, Etienne J, Chidiac C.
In vivo effect of adhesion inhibitor heparin on Legionella pneupophila
pathogenesis in a murine pneumonia model.
Intensive Care Med, 2008, 34:1511-1519.
Mascart F, Hainaut M, Peltier A, Verscheure V, Levy J, Locht C.
Modulation of the infant immune responses by the first pertussis vaccine
adminstrations.
Vaccine, 2007, 25:391-398.
Allix−Béguec C, Fauville−Dufaux M, Supply P.
Three-year population-based evaluation of standardized Mycobacterial
Interspersed Repetitive Unit-Variable Number of Tandem Repeat typing
of Mycobacterium tuberculosis.
J Clin Microbiol, 2008, 46:1398-1406.
Niemann S, Weniger T, Harmsen D, Supply P.
Mycobacteria MIRU-VNTRplus : Online database for identification of M.
tuberculosis complex isolates based on MIRU, SPOLIGO, and regions of
difference data.
Internat J Med Microbiol, 2007, 297:149-149 Suppl. 43.
Allix-Béguec C, Harmsen D, Weniger T, Supply P, Niemann S.
Evaluation and strategy for use of MIRU-VNTRplus, a multifunctional
database for online analysis of genotyping data and phylogenetic
identification of Mycobacterium tuberculosis complex isolates.
J Clin Microbiol, 2008, 46:2692-2699.
Oelemann MC, Diel R, Vatin V, Haas W, Rusch-Gerdes S, Locht C,
Niemann S, Supply P.
Assessment of an optimized Mycobacterial Interpersed Repetitive UnitVariable Number of Tandem Repeat typing system combined with
spoligotyping for population-based molecular epidemiology studies of
tuberculosis.
Allix−Béguec C, Supply P, Wanlin M, Bifani P. Fauville−Dufaux M.
Standardised PCR−based molecular epidemiology of tuberculosis in the
Brussels Capital Region.
Eur Resp J, 2008, 31:1077-1084.
Rucktooa P, Antoine R, Herrou J, Huvent I, Locht C, JacobDubuisson F, Villeret V, Bompard C.
Crystal structures of two Bordetella pertussis periplasmic receptors
contribute to defining a novel pyroglutamic acid binding DctP subfamily.
J Mol Biol, 2007, 370:93-106.
Alsteens D, Verbelen C, Dague E, Raze D, Baulard,AR.
Organization of the mycobacterial cell wall : a nanoscale view.
Pflugers Arch, 2008, 456:117-125.
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Amniai L, Biet F, Marquillies P, Locht C, Pestel J, Tonnel AB, Duez C.
IL-18 does not increase allergy airway disease in mice when prooduced
by BCG.
J Biomed Biotechnol, 2007, 2007:67276.
Thibault VC, Grayon M, Boschiroli ML, Willery E, Allix-Béguec C,
Stevenson K, Biet F, Supply P.
Combined multilocus short-sequence-repeat and mycobacterial
interspersed repetitive unit-variable-number tandem-repeat typing of
Mycobacterium avium subsp. pratuberculosis isolates.
J Clin Microbiol, 2008, 46:4091-4094.
Coutte L, Botkin DJ, Gao L, Norris SJ.
Detailed analysis of sequence changes occuring during vlsE antigenic
variation in the mouse model of Borrelia burgdorferi infection.
PLoS Pathog, 2009, 5:e1000293.
Verbelen C, Dupres V, Raze D, Bompard C, Locht C, Dufrêne YF.
Interaction of the mycobacterial heparin-binding hemagglutinin with
actin, as evidenced by single-molecule force spectroscopy.
J Bacteriol, 2008, 190:7614-7620.
Dé E, Saint N, Glinel K, Méli A, Lévy D, Jacob-Dubuisson F.
Influence of the passenger domain of a model autotransporter on the
properties of its translocator domain.
Mol Membr Biol, 2008, 25:192-202.
Wirth T, Hildebrand F, Allix-Béguec C, Wölbeling F, Kubica T,
Kremer K, van Soolingen D, Rüsch-Gerdes S, Locht C, Brisse S,
Meyer A, Supply P, Niemann S.
Origin, spread and demography of the Mycobacterium tuberculosis
complex.
PLoS Pathog, 2008, 4:e1000160.
FeunouFeunou P, Ismaili J, Debrie AS, Huot L, Hot D, Raze D,
Lemoine Y, Locht C.
Genetic stability of the live attenuated Bordetella pertussis vaccine
candidate BPZE1.
Vaccine, 2008, 26:5722-5727.
Wohlkönig A, Hodak H, Clantin B, Sénéchal M, Bompard C, JacobDubuisson F, Villeret V.
Crystallization and preliminary X-ray diffraction analysis of the
peptidylprolyl isomerase Par27 of Bordetella pertussis.
Acta Crystallogr Sect F Struct Biol Cryst Commun, 2008, 64:809-812.
Guerrero G, FeunouFeunou P, Locht C.
The coiled-coil N-terminal domain of the mycobacterial heparin-binding
haemagglutinin (HBHA) is required for the humoral and cellular immune
responses in mice.
Mol Immunol, 2008, 46:116-124.
Wolowczuk I, Verwaerde C, Viltart O, Delanoye A, Delacre M, Pot B,
Grangette C.
Feeding our immune system : impact on metabolism.
Clin Dev Immunol, 2008, 639803.
Ho SY, Chua SQ, Foo DGW, Locht C, Chow VT, Poh CL, Alonso S.
The highly attenuated Bordetella pertussis BPZE1 strain as a potential live
vehicle for the delivery of heterologous vaccine candidates.
Infect Immun, 2008, 76:111-119.
2009
Hodak H, Wohlkönig A, Smet-Nocca C, Drobecq H, Wieruszeski JM,
Sénéchal M, Landrieu I, Locht C, Jamin M, Jacob-Dubuisson F.
The peptidyl-prolyl isomerase and chaperone Par27 of Bordetella
pertussis as the prototype for a new group of parvulins.
J Mol Biol, 2008, 376:414-426.
Baud C, Hodak H, Willery E, Drobecq H, Locht C, Jamin M, JacobDubuisson F.
Role of DegP for two-partner secretion in Bordetella.
Mol Microbiol, 2009, 74:315-329.
Delbende C, Verwaerde C, Mougel A, Tranchand Bunel D.
Induction of therapeutic antibodies by vaccination against external loops
of tumor-associated viral latent membrane protein.
J Virol, 2009, 83:11734-11745.
Locht, C.
A common vaccination strategy to solve unsolved problems of
tuberculosis and pertussis ?
Microbes Infect, 2008, 10:1051-1056.
Dirix V, Verscheure V, Goetghebuer T, Hainaut M, Debrie AS,
Locht C, Mascart F.
Monocyte-derived interleukin-10 depresses the Bordetella pertussis
specific gamma interferon response in vaccinated infants.
Clin Vaccine Immunol, 2009, 16:1816-1821.
Locht C, Raze D, Rouanet C, Genisset C, Segers J, Mascart F.
The Mycobacterial Heparin-Binding Hemagglutinin : a Virulence Factor
and Antigen Useful for Diagnostics and Vaccine Development. In The
Mycobacterial cell Enveloppe.
Edited by M. Daffé and J-M Reyrat. 2008 ASM Press, Washington, DC.
Dirix V, Verscheure V, Goetghebuer T, Hainaut M, Debrie AS, Locht
C, Mascart F.
Cytokine and antibody profiles in 1-year-old children vaccinated with
either acellular or whole-cell pertussis vaccine during infancy.
Vaccine, 2009, 27:6042-6047.
Poulain-Godefroy O, Vendeville C, Locht C, and Riveau, G.
Bordetella pertussis filamentous haemagglutinin delivered by mucosal
routes enhances immunoglobulin level in serum and mucosal fluids.
FEMS Immunol Med Microbiol, 2008, 54:129-136.
Simonney N, Dewulf G, Herrmann JL, Gutierrez MC, Vicaut E,
Boutron C, Leportier M, Lafaurie M, Roux X, Dubuquoy C, Durand
G, Tran-Tolla TL, Castagné N, Bernard J, Petit-Camurdan A, Eléouet
J F, Riffault S.
Sub-nucleocapsid nanoparticles : a nasal vaccine against respiratory
syncytial virus.
PLoS One 3, 2008, e1766.
Doherty M, Wallis RS, Zumla A; WHO-Tropical Disease
Research/European Commission joint expert consultation group.
Biomarkers for tuberculosis disease status and diagnosis.
Curr Opin Pulm Med, 2009, 15:181-187.
Gutierrez MC, Supply P, Brosch R.
Pathogenomics of Mycobacteria.
Genome Dyn, 2009, 6:198-210.
Supply P.
MIRU-VNTRplus (http://www.miru-vntrplus.org) : Multifunctional data
base for on-line analysis of genotypes and phylogenetic identification of
the Mycobacterium tuberculosis complex strains.
Developed in collaboration with Dag Harmsen, Thomas Weniger
(University of Münster, D), et Stefan Niemann (Borstel Research Centre,
D). 2008.
Herrou J, Debrie AS, Willery E, Renaud-Mongénie G, Locht C, Mooi
F, Jacob-Dubuisson F, Antoine R.
Molecular evolution of the two-component system BvgAS involved in
virulence regulation in Bordetella.
PLoS One, 2009, 4:e6996.
31
Contents
Jacob-Dubuisson F, Villeret V, Clantin B, Delattre AS, Saint N.
First structural insights into the TpsB/Omp85 superfamily.
Biol Chem, 2009, 390:675-684.
Delattre AS, Cantin B, Saint N, Locht C, Villeret V, JacobDubuisson F.
Functional importance of a conserved sequence motif in FhaC, a
prototypic member of the TpsB/Omp85 superfamily
FEBS J, 2010, in press.
Juffroy O, Noël D, Delanoye A, Viltart O, Wolowczuk I, Verwaerde C.
Subcutaneous graft of D1 mouse mesenchymal stem cells leads to the
formation of a bone-like structure.
Differentiation, 2009, 78:223-231.
Feunou PF, Kammoun H, Debrie AS, Mielcarek N, Locht C.
Long-term immunity against pertussis induced by a single nasal
administration of live attenuated B. pertussis BPZE1.
Vaccine, 2010, in press.
Lechner M, Schmitt K, Bauer S, Hot D, Hubans C, Levillain E, Locht
C, Lemoine Y, Gross R.
Genomic island excisions in Bordetella petrii.
BMC Microbiol, 2009, 18:141.
Feunou PF, Bertout J, Locht C.
T- and B-cell-mediated protection induced by novel, live attenuated
pertussis vaccine in mice. Croo protection against parapertussis.
PLoS One, 2010, 5:e10178.
Mathys V, Wintjens R, Lefevre P, Bertout J, Singhal A, Kiass M,
Kurepina N, Wang XM, Mathema B, Baulard A, Kreiswirth BN,
Bifani P.
Molecular genetics of para-aminosalicyclic acid resistance in clinical
isolates and spontaneous mutants of Mycobacterium tuberculosis.
Antimicrob Agents Chemother, 2009, 53:2100-2109.
Guerrero GG, Debrie AS, Locht C.
Boosting with mycobacterial heparin-binding haemagglutinin enhances
protection of Mycobacterium bovis BCG-vaccinated newborn mice
against M. tuberculosis.
Vaccine, 2010, 28:4340-4347.
Rouanet C, Debrie AS, Lecher S, Locht C.
Sybcutaneous boosting with heparin binding haemagglutinin increases
BCG-induced protection against tuberculosis.
Herrou J, Bompard C, Wintjens R, Dupré E, Willery E, Villeret V,
Locht C, Antoine R, Jacob-Dubuisson, F.
The periplasmic domain of the sensor-kinase BvgS reveals a new
paradigm for the Venus flytrap mechanism.
Proc Natl Acad Sci USA, 2010, in press.
Skerry CM, Cassidy JP, English K, Feunou-Feunou P, Locht C,
Mahon BP.
A live attenuated Bordetella pertussis candidate vaccine does not cause
disseminating infection in gamma interferon receptor knockout mice.
Kavanagh H, Noone C, Cahill E, English K, Locht C, Mahon BP.
Attenuated Bordetella pertussis vaccine strain BPZE1 modulates allergeninduced immunity and prevents allergic pulmonary pathology in a
murine model.
Clin Exp Allgery, 2010, 40:933-941.
Verbelen C, Christiaens N, Alsteens D, Dupres V, Baulard AR,
Dufrêne YF.
Molecular mapping of lipoarabinomannans on mycobacteria.
Langmuir, 2009, 25:4324-4327.
Li R, Lim A, Phoon MC, Narasaraju T, Ng JK, Poh WP, Sim MK, Chow
VT, Locht C, Alonso S.
Attenuated Bordetella pertussus protects against highly pathogenic
influenza A viruses by dampening the cytokine storm.
J Virol, 2010, 84:7105-7113.
Willand N, Dirié B, Carette X, Bifani P, Singhal A, Desroses M,
Leroux F, Willery E, Mathys V, Déprez-Poulain R, Delcroix G,
Frénois F, Aumercier M, Locht C, Villeret V, Déprez B, Baulard AR.
Synthetic EthR inhibitors boost antituberculous activity of ethionamide.
Nat Med, 2009, 15:537-544.
Mielcarek N, Debrie AS, Mahieux S, Locht C.
Dose-response of attenuated Bordetella pertussis BPZE1-induced
protection in mice.
Leroux F, Willery E, Mathys V, Déprez- Dupres V, Verbelen C, Raze D,
Lafont F, Dufrêne YF.
Force spectroscopy of the interaction between mycobacterial adhesins
and heparan sulphate proteoglycan receptors.
Chemphyschem, 2009, 10:1672-1675.
Narayanan S, Swaminathan S, Supply P, Shanmugam S, Narendran G,
Hari L, Ramachandran R, Locht C, Jawahar MS, Narayanan PR
Impact of HIV infection of the Recurrence of Tuberculosis in South India.
J Infect Dis, 2010, 201:691-703.
2010
Place S, Verscheure V, de San N, Hougardy JM, Schepers K, Dirix V,
Dediste A, Michel O, Drowart A, Allart SD, Doherty TM, Lecher S,
Locht C, Mascart F.
Heparin-binding, Hemagglutinin-specific IFN-(gamma). Synthesis at the
Site of Infection during Active Tubersulosis in Humans.
Al-Hajoj SA, Akkerman O, Parwati I, Al-Gamdi S, Rahim Z,
van Soolingen D, van Ingen J, Supply P, van der Zanden AG.
Micro-evolution of Mycobacterium tuberculosis in a tuberculosis patient.
J Clin Microbiol, 2010, in press.
Allix-Béguéc C, fauville-Dufaux M, Stoffels K, Ommeslag D,
Walravens K, Saegerman C, Supply P.
Importance of identifying Mycobacterium bovis as a causative agent of
human tuberculosis.
Eur Respir J, 2010, 35:692-694.
Radomski N, Thibault VC, Karoui C, de Cruz K, Cochard T,
Gutiérrez C, Supply P, Biet F, Boschiroli ML.
Determination of genotypic diversity of Mycobacterium avium subspecies
from human and animal origns by mycobacterial interspersed repetitiveunit-variable-number tandem-repeat and IS1311 restriction fragment
length polymorphism typing methods.
J Clin Mrirobiol, 2010, 3:339-348.
Clantin B, Leyrat C, Wohlkönig A, Hodak H, Ribeiro ED Jr,
Martinez N, Baud C, Smet-Nocca C, Villeret V, Jacob-Dubuisson F,
Jamin M.
Structure and plasticity of the peptidyl-prolyl isomerase Par27 of
Bordetella pertussis revealed by X-rat diffraction and small X-ray
scattering.
J Struct Biol, 2010, 169:253-265.
Rouanet C, Locht C.
Boosting BCG to protect against TB.
Expert Rev Respir Med, 2010, 3:339-348.
32
Contents
Patents
Shamputa IC, Lee J, Allix-Béguec C, Cho EJ, Lee JI, Rajan V, Lee EG,
Min JH, Carroll MW, Goldfeder LC, Kim JH, Kang HS, Hwang S,
Eum SY, Park SK, Lee H, Supply P, Cho SN, Via LE, Barry CE 3rd.
Geentic diversity of Mycobacterium tuberculosis isolates from a tertiary
care tuberculosis hospital on South Korea.
Locht C, Mahon B, Kavanagh H.
Vaccine for prophylaxis or treatment of an allergen-driven airway
pathology
April 28, 2009 (EP 09 305 371.8)
IPL, Inserm, National University of Ireland, Maynooth
Radomski N, Thibault VC, Karoui C, de Cruz K, Cochard T,
Gutiérrez C, Supply P, Biet F, Boschiroli ML.
Genotypic diversity of Mycobacterium avium subscpecies from human
and animal origins, studied by MIRU-VNTR and IS1311 RFLP typing
methods.
J Clin Microbiol, 2010, in press.
Locht C, Roux X, Kammoun H, Raze D.
Use of Pal1 as carrier for heterologous antigen presentation by a life
gram-negative bacterial vaccine
April 28, 2009 (EP 09 305 370.0)
IPL, Inserm
Vermeulen F, Verscheure V, Damis E, Vermeylen D, Leloux G, Dirix V,
Locht C, Mascart F.
Cellular immune responses of preterm infants after vaccination with
whole-cell or acellular pertussis vaccines.
Clin Vaccine Immunol, 2010, in press
Locht C, Alonso S, Chow V, Li R.
Influenza vaccine, composition, and methods of use
June 16, 2009 (PCT/US09/47399)
IPL, Inserm, National University of Singapore
Weniger T, Krawczyk J, Supply P, Niemann S, Harmsen D.
MIRU-VNTRplus : a web tool for polyphasic genotyping of Mycobacterium
tuberculosis complex bacteria.
Nucleic Acids Res, 2010, 38 Suppl:W326-331.
Willand N, Desroses M, Toto P, Dirié B, Lens Z, Villeret V, Rucktooa P,
Locht C, Baulard A, Deprez B.
Exploring drug target flexibility using in Situ Click Chemistry : Application
to a Mycobacterial transcriptional regulator.
ACS Chem Biol, 2010, in press.
PhD
Hélène HODAK
Directeur de thèse : Camille Locht
Les partenaires et interactions périplasmiques au cours de la sécrétion de
l’hémagglutinine filamenteuse par la voie à deux partenaires chez
Bordetella pertussis
08 juin 2007
Julien HERROU
Directeur de thèse : Françoise Jacob-Dubuisson et Rudy Antoine
Etude du régulateur central de la virulence, BvgA/S chez Bordetella
pertussis, l’agent de la coqueluche
03 décembre 2008
Xavier CARETTE
Directeur de thèse : Alain Baulard
Etude du contrôle de la bio-activation de l’éthionamide et applications
thérapeutiques antituberculeuses
30 septembre 2009
33
Contents
how is it regulated? Which molecular components from the
host and from the pathogen can be targeted for designing
new therapeutic tools interfering with this early signaling
response activated at the interface level? These are the
questions the team addresses by using several pathogen
models that have evolved different adhesion and invasion
strategies. The team develops a translational project in which
several pathogens are used as experimental models in order
to analyze the similar and specific molecular mechanisms
involved. The main point herein is that investigations are
focused at the interface between the pathogen and the host
cell membranes.
Cellular Microbiology
of Infectious Pathogens
Frank LAFONT, DR2 CNRS
Group Members :
Bertout Julie, CE IPL (congès CIF)
Ciczora Yann, IE CNRS
Pomel Sébastien, Postdoctoral fellow IPL-Région
Smaali Nassima, Postdoctoral fellow CNRS
Warein Joëlle, Technician IPL
1. The membrane-associated
signaling pathway during the first
steps of Shigella flexneri invasion
Key words :
Host-pathogen interaction. Cell biology. Biophysics.
Cell signaling.
We are interested in identifying signaling molecules that could
be recruited to the host cell membranes (at the cell surface
and vacuolar membrane levels) during the early steps of
Shigella host cell invasion. As a first approach, we
characterized the ubiquitinylation pathway. We reasoned that
many signaling molecules are targets for ubiquitinylation. This
led us to discriminate between mono-, multi- and polyubiquitinylated proteins. We obtained data for a massive
ubiquitinylation occuring on vacuolar membranes after
disruption that drives a sustained signaling activation until the
membranes are trapped by the autophagy pathway. On these
membranes, we identified molecular components of
important signaling pathways leading to an inflammatory
response (sequestosome, inflammasome) and to cell death
(pyroptosis inducer caspase-1). This study addressed the
original issue of the signaling associated with the remnant
membranes left after the escape of the bacterium to the
cytoplasm. We unveiled an underestimated signaling
regulation at the vacuolar membrane that will open new
avenues for a better understanding of how the cell response
is orchestrated by intracellular invading bacteria like Shigella
and Listeria.
In the course of this study we made use of an observation
obtained in Sansonetti’s laboratory at the Pasteur Institute in
Paris that shows the recruitment of galectin-3 as soon as the
vacuolar membrane breaks. We were able to monitor the
recruitment of Gal-3 by videoTIRF and deciphered the
recruitment mechanism. We provided evidence that the
glycosylated residues of the surface host molecules that were
exposed at the cell surface, or were intraluminal in
the vacuole, and that became accessible in the cytoplasm
after vacuolar membrane disruption, recruit the cytoplasmic
galectin-3.
A - Research Report
Frank Lafont inaugurated his team at the Pasteur Institute of
Lille the 26th of September, 2005. The laboratory was initially
equipped by grants form the French Minister for Research and
Higher Education (Chaire d’Excellence) and from the charity
foundation “Fondation pour la Recherche Médicale”. In 2005
and 2007, grants from the National Research agency were
obtained as well as an ARCI program from the Région Nord
Pas-de-Calais in 2008 and in 2006 the team obtained the
“Fondation pour la Recherche Médicale” designation. The
team has introduced several new techniques on the Lille
Pasteur campus like atomic force microscopy (AFM) and total
internal reflection fluorescence (TIRF) microscopy.
During his post-doctoral training at the European Molecular
Biology Laboratory in Heidelberg in Kai Simon’s team F. Lafont
worked on the polarity, dynamics and compartmentalization
of membrane microdomains. He became interested in the
hijacking of lipid rafts by bacterial pathogens during his work
as a junior lecturer in G van der Goot’s laboratory at the
University of Geneva. In particular, he demonstrated that
Shigella flexneri subverted rafts for the entry step and that the
type three secretion system of S. flexneri interacts directly with
raft molecular components. Also, while in Switzerland, he
initiated an original research program on the use of AFM to
investigate the biophysical properties of the plasma
membrane in living cells with colleagues at the Swiss Federal
Institute of Technology at Lausanne. This was the basis of a
new line of research based on an interdisciplinary approach
combining bacterial genetics, biochemistry, cellular and
molecular biology, imaging and biophysics.
The working hypothesis of the team is that the mechanical
properties of the plasma membrane influence the dynamics
of the signaling cascade triggered by the adhesion of
pathogens on cell membranes. One main original idea is that
there is a basic component common to many adhesion and
invasion phenomena that is hindered by specific interactions
(that may rely on high and/or low affinities). By adhering to
the cell surface, pathogens define specialized membrane
domains. How does this confinement influence the cell
signaling response? What is the molecular basis of the
mechanical changes? How long is the signal maintained and
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Contents
Altogether, these data led to the working hypothesis that
septins can regulate the subcortical network on the
cytoskeleton onto which the receptor is anchored. As such,
this work using an original approach in the field gives a clue
into the physiological function of septins.
Infected cells labeled for polyubiquitin, Gal-3, bacteria and filipin.
Mapping of the elastic Young modulus on the topogram
of a cell with indication by red arrows
of the InlB-Met interaction sites.
2. The role of the septin cytoskeleton
in Listeria monocytogenes invasion
The detailed mechanism allowing Listeria monocytogenes to
invade the host cells awaits comprehensive characterization.
Listeria uses the zipper mode of entry at variance with the
triggered mode used by Shigella. However, both pathogens
end up in the cytoplasm where they use an actin-based
motility mechanism to invade the neighboring cells.
Moreover, contrary to Shigella that binds loosely to the host
cell surface, Listeria binds tightly to two receptors. Hence, the
Listeria model offers a complementary experimental model
to investigate the pathogen-host membrane interface. We are
interested in the molecular basis of the adhesion of Listeria to
the host cell surface and on the subsequent signaling
response at the interface. We decided first to examine the
mechanical features underlying the adhesion step. Listeria’s
membrane receptors are anchored to the subcortical network
of actin. In this context we wanted to investigate the role
played by the septin family. The function of septins remains
unclear, although they are involved in the regulation of the
cytoskeleton and in the membrane-cytoskeleton
relationships. We have shown in collabration with the P
Cossart’s Team at the Pasteur Insitute in Paris, using atomic
force microscopy (AFM), that depletion of septin family
members leads to different cell shapes, either bigger or
smaller than those of control cells, especially along the z-axis.
Interestingly, invasion of Listeria monocytogenes into cells
depleted for septins is differently affected depending on the
family member targeted and is correlated with the cell shape.
Also using AFM, we have shown that the interaction between
one Listeria adhesin, InlB, and its receptor, Met, is also affected
and correlated with the cell shape. By targeting some
members of the septin family it is possible to modify the
interaction forces between InlB and its receptor. As our
measurements were performed on living cells, the interaction
forces recorded take into account both the molecular
interaction and the elasticity of the cell. Hence, our approach
gave a hint concerning the anchorage of the receptor on the
cytoskeleton and on the mechanical properties of the
membrane engaged in adhesion to the microorganism.
3. The internalization pathway
of Yersinia pseudotuberculosis
in host cells
Although Yersinia is described as being bound at the cell
surface, where it can replicate, there are reports, both in vivo
and in vitro, arguing for Yersinia internalization into
macrophages. This pathogen, like Listeria, uses the zipper
mode of invasion but remains intravesicular. With Yersinia, we
are interested in identifying the molecular signaling
machineries that are recruited at the host cell membrane
level while the bacteria are internalized and replicating
(interestingly bacteria remain closely associated with the
vacuolar membrane as observed by electron microscopy).
However, the membrane trafficking of the microorganism
remained unclear and we first aimed at deciphering the entry
pathway. We showed in collaboration with M. Simonet’s Team
at the Institut Pasteur de Lille that Y. pseudotuberculosis
hijacked membrane lipid rafts during its entry into the host
cells. Then, the pathogen subverted the autophagy pathway
as shown using specific markers and functional approaches
based on the expression of dominant inhibitory mutants of
this pathway. Moreover, we could establish a link between
activation of autophagy and inhibition of apoptosis, which
allows successful bacterial invasion. However, cell death is
observed at later stage of infection and we could
demonstrate that the binding of the bacterium to the cell
surface triggered activation of pyroptosis that ultimately led
to cell death and bacterial release.
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Contents
5. Interaction of LPS with the host cell
surface
The LPS of Gram-negative bacteria is composed of a
hydrophobic lipid A linked to a charged, densely compact
oligosaccharidic core associated (smooth-LPS) or not (roughLPS) with a hydrophilic O-polysaccharide chain (O-chain).
LPS protects enteroinvasive bacteria against the
inflammatory reaction and one of the bacterial elements that
encounter the host is LPS. Interaction between LPS and the
host cell plasma membrane may be responsible for
stabilizing the bacterium onto the plasma membrane of the
host. Using signature-tagged mutagenesis, genes invoved in
the glucosylation of the tri-rhamnose-N-acetyl glucosamine
tetrasaccharide O antigen of Shigella were shown to regulate
the size of Shigella LPS molecule and regulate Shigella
virulence. Therefore, it is important to check the influence of
purified LPS in the reorganization of the host plasma
membrane. Indeed, several lines of evidence suggest that
Brucella LPS, by itself is an important modulator of
membrane domains. Brucella possesses a peculiar LPS that
we call non-classical LPS as compared with the so-called
classical LPS from enterobacteria such as Escherichia coli. B.
abortus lipid A possesses a diaminoglucose backbone
(rather than glucosamine), and acyl groups are longer (C18–
C19, C28 rather than C12 and C14) and are only linked to the
core by amide bounds (rather than ester and amide bonds).
In collaboration with JP. Gorvel’s Group at the CIML in
Marseille, we thus wanted to characterize the interaction
forces of several LPS from Brucella with the host plasma
membrane of living cells. However, there is no method
available for LPS coating onto the AFM tip. We have thus first
established a method allowing targeting of the acyl chains
of lipid A for tip functionalisation that will be then used in
the field.
Infected cells labeled for tubulin, DNA, autophagosomes
and gangliosides
We also have investigated the role of the microbiota flora in
protection against Y. pseudotuberculosis and have shown that
MyD88- and TLR2-dependent sensing is instrumental in
eliciting degranulation of Paneth cells.
4. The role of knobs ultrastructure
and elasticity in regulating
cytoadherence
Once infected by Plasmodium falciparum, red blood cells
express on their cell surface specific membrane structures
connected to the underlying cytoskeleton that result from
parasite protein synthesis. Some of these structures, known
as knobs, are assumed to play an important role in the
cytoadherence of the infected red blood cells to the vascular
wall. With this experimental model, in collaboration with A.
Scherf’s Team at the Pasteur Institute in Paris, we can
investigate how a pathogen can modulate the mechanical
properties of the host cell that in turn influence
cytoadherence. We first aimed at characterizing, in a
biophysical perspective, these membrane structures induced
by the parasite. We have thus been able, using AFM, to
characterize the ultrastructure of these knobs as a function of
mutations and to correlate some phenotypic differences
observed with different genotypes. The genotypes that gave
the strongest differences as revealed with AFM were then
used for further analysis of cell elasticity.
6. Mechanical adhesion properties
of bacilli and bacteria
To better understand the microorganism - host cell interface,
we investigate the adhesion properties of the microorganism
in a simple experimental system, without the eukaryotic cell,
in order to characterize some basic phenomena. We selected
as experimental systems the adhesion of bacilli to inert
surfaces, such as those used in food industries, and biofilm
formation onto such substrata. This offered the possibility of
investigating the mechanical features of pathogenic
microorganisms (e.g. Bacilus anthracis and Listeria) and to
address questions that are of high relevance for industry, i.e.
contamination by adhering microorganisms. This project as
part of a consortium with groups from the INRA, AFSSA,
INSERM and INSA, launched during the second semester of
2008 aims at the characterization, using AFM, of the adhesion
properties of spores (with B. Cereus as the experimental model
for bacilli) onto metal substrata and biofilms (with Listeria as
the model for contamination of fish products). Moreover, AFM
will be also used to test on living cells the adhesion properties
of bacilli expressing a mutant of BclA (Bacillus collagen-like
protein of anthracis) that is localized in the exosporium and
plays a role in bacilli adhesion. Conversely, tips functionalized
AFM images of infected erythrocytes
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Contents
with BclA will be used directly on living cells to measure
interaction forces in correlation with the local elasticity of the
cell membrane.
2009
Dessein R, Gironella M, Vignal C, Peyrin-Biroulet L, Sokol H,
Secher T, Lacas-Gervais S, Gratadoux JJ, Lafont F, Dagorn JC,
Ryffel B, Akira S, Langella P, Nùñez G, Sirard JC, Iovanna J,
Simonet M, Chamaillard M.
TLR2 is critical for induction of REG3{beta} expression and intestinal
clearance of Yersinia pseudotuberculosis.
Gut, 2009, 58:771-776.
Dupont N, Lacas-Gervais S, Bertout J, Paz I, Freche B, Van Nhieu GT,
van der Goot FG, Sansonetti PJ, Lafont F.
Shigella phagocytic vacuolar membrane remnants participate in the
cellular response to pathogen invasion and are regulated by autophagy.
Cell Host Microbe, 2009, Aug 20;6:137-149.
Dupont N, Lafont F.
How autophagy regulates the host cell signaling associated with the
postpartum bacteria cocoon experienced as a danger signal.
Autophagy, 2009, 5:1222-1223.
Dupres V, Verbelen C, Raze D, Lafont L, Dufrêne YF.
Force spectroscopy of the interaction between mycobacterial adhesions
and heparan sulphate proteoglycan receptors Chem Phys
Chem, 2009, 10:1672-1675.
AFM imaging of B cereus with exosporium
Roduit C, Dietler G, Sekatski S, Catsicas S, Lafont L, Kasas S.
Stiffness tomography by atomic force microscopy.
Biophys J, 2009, 97:674-677.
development 2010
2010
Lafont F.
Autophagy employs a new DAGger against bacteria.
Cell Host Microbe, 2010, 8:129-130.
Our project for the considered period will be to extend our analysis of
the membrane-associated signalling activation triggered upon
interaction of pathogens with the host cell membranes, based on our
previous results and technological achievements. We previously found
that membrane signalling can affect host cell survival, once infected,
and the membrane traffic of different pathogens. Hence, we want to
challenge our working hypothesis that upon pathogen interaction, the
pathogen – host cell membrane interface defines membrane domains
where recruited signalling molecules initiate pathways that regulate
survival of the host and/or survival/replication of the pathogen. Our
methodological strategy remains based on a multidisciplinary approach
taking advantages of a new technological development related to the
coupling of high-resolution biophotonic microscopy, to analyze the
distribution of signalling molecules, and Atomic Force Microscopy
(AFM), to examine the biophysical properties of the membrane into
which the signalling molecules are recruited and activated.
Moreau K, Lacas-Gervais S, Fujita N, Sebbane F, Yoshimori T,
Simonet M, Lafont F.
Autophagosomes can support Yersinia pseudotuberculosis replication in
macrophages.
Cell Microbiol, 2010, 12:1108-1123.
Paz I, Sachse M, Dupont N, Cederfur C, Enninga J, Leffler H,
Poirier F, Prevost MC, Lafont F, Sansonetti P.
Galectin-3, a marker for vacuole lysis by invasive pathogens.
Cell Microbiol, 2009, 27 nov ahead of print.
PhD
Kévin Moreau
Directeur de thèse : Frank Lafont
« Caractérisation fonctionnelle des voies cellulaires détournées lors de
l’infection in vitro par les Yersinia »
Université de Lille 2 - Faculté de Médecine
21 janvier 2009
Publications
2007
Nicolas Dupont
Directeur de thèse : Frank Lafont
“Caractérisation des voies de signalisation associées aux membranes au
stade précoce de l'infection par Shigella flexneri »
Université de Lille 2 - Faculté de Médecine
26 août 2009
Yersin A, Hirling H, Kasas S, Roduit C, Kulangara K, Dietler G, Lafont
F, Catsicas S, Steiner P.
Elastic properties of the cell surface and trafficking of single AMPA
receptors in living primary hippocampal neurons.
Biophys J, 2007, 92:4482-4489.
2008
Roduit C, van der Goot FG, De Los Rios P, Yersin A, Steiner P, Dietler
G, Catsicas S, Lafont F, Kasas S.
Elastic membrane heterogeneity of living cells revealed by stiff nanoscale
membrane domains.
Biophys J, 2008, 94:1521-1532.
37
Contents
which we contributed. These pseudoparticles consist of fulllength HCV envelope glycoproteins assembled onto retroviral
core particles containing a retrovirus-derived genome
harboring a marker gene. More recently, the development of a
cell culture system that allows for a relatively efficient
amplification of HCV (HCVcc) has finally been reported. This
system is based on the transfection of the human hepatoma
cell line Huh-7 with genomic HCV RNA derived from a cloned
viral genome of an HCV isolate from a Japanese patient with
fulminant hepatitis (JFH-1 isolate). HCVcc is now the most
relevant system to study the HCV life cycle.
The main objective of the team in the past 4 years was to study
some of the molecular and cellular aspects of the interactions
between HCV and its major target cell, the hepatocyte. The
team has a long-standing experience in studying HCV envelope
glycoproteins and the cellular aspects of HCV entry, and most
of its projects have been focused on these topics.
Molecular and Cellular
Virology of Hepatitis C
Jean DUBUISSON, DR1 CNRS
Group Members :
Belouzard Sandrine, CR2 CNRS
Cocquerel Laurence, CR1CNRS
Rouillé Yves, DR2 CNRS
Séron Karin, CR1CNRS
Wychowski Czeslaw, DR2 CNRS
Callens Nathalie, IE2 CNRS
Montpellier Claire, IE2 CNRS
Pillez André, Technician CNRS
Ung Sophana, Technician IPL
Popescu Costin-Ioan, Postdoctoral fellow CNRS
Meuleman Philip, Postdoctoral fellow FWO (Belgium)
Tews Birke, Postdoctoral fellow Marie Curie
Thomas Xavier, Postdoctoral fellow CNRS
Albecka Anna, PhD Student CNRS
Aslaheh Khaled, PhD Student CNRS
Calland Noémie, PhD Student MENRT
Delavalle Pierre Yves, PhD Student MENRT
Goueslain Lucie, PhD Student ANRS
Potel Julie, PhD Student MENRT
Vieyres Gabrielle, PhD Student MENRT
1. Functional studies of HCV
envelope glycoproteins
The genome of HCV encodes a single polyprotein of about 3000
amino acids, which is cleaved co- and post-translationally by
cellular and viral proteases to yield at least ten mature products.
Cleavage of the viral polyprotein by a cellular signal peptidase
gives rise to the envelope glycoproteins, E1 and E2. Our
laboratory has been a pioneer in characterizing the biogenesis
of HCV envelope glycoproteins. Briefly, these glycoproteins are
type I membrane proteins containing a large N-terminal
ectodomain and a C-terminal transmembrane domain. During
their synthesis, E1 and E2 ectodomains are directed to the lumen
of the endoplasmic reticulum (ER), and their transmembrane
domains are inserted into the membrane of this compartment.
During their biogenesis, E1 and E2 assemble as non-covalent
heterodimers which are retained in the ER. These heterodimers
are the form of E1E2 present at the surface of viral particles and,
thus, must be involved in interaction(s) with cellular receptor(s).
Furthermore, E1 present at the surface of HCV particles has also
been shown to be trimeric, suggesting that HCV envelope
glycoproteins form a network of interactions that is probably
important in the assembly and entry steps of the HCV life cycle.
After having deeply characterized the transmembrane domains
of HCV envelope glycoproteins, we have also compared the
transmembrane domains with those of the envelope proteins
of yellow fever virus, another member of the Flaviviridae family.
Interestingly, our data have indicated that the organization of
these domains and their mechanism of ER localization are
different in yellow fever virus as compared to HCV.
Besides the biogenesis of HCV envelope proteins, we have also
characterized the role of some regions of these proteins in HCV
entry. During the past four years, we have mainly focused
our activity on the hypervariable region 1 (HVR1) of HCV
glycoprotein E2, on the glycans associated with HCV envelope
proteins and on the transmembrane domains. HVR1 is located
at the N-terminus of E2 and has been proposed to modulate
the accessibility to some entry factors. This region has been
described as globally basic, with positively charged residues
located at specific sequence positions. We have demonstrated
that HCV entry globally increases with the number of basic
residues in HVR1, confirming the role of these residues in HCV
entry. We have also focused some of our research activities on
Key words :
HCV, Virus entry. HCV envelope. Viral glycoproteins. Virus-cell
interactions. Virus assembly. Virus life cycle. Flaviviridae.
A - Research Report
The MCVHC is currently part of the CNRS UMR 8161 (Institut de
Biologie de Lille). Hepatitis C virus (HCV) is a major cause of
chronic hepatitis, cirrhosis and hepatocellular carcinoma
throughout the world, which is due to the fact that viral replication occurs primarily, if not exclusively, within hepatocytes. HCV
has also become the main indication for liver transplantation in
a number of countries. Interferon alpha and the nucleoside
analogue ribavirin form the basis for treatment, but have
numerous side effects and are not sufficiently effective, especially against genotype 1, one of the most frequent genotypes
in industrialized countries. With an estimated 170 million
people infected, i.e. nearly 3% of the world population, and an
incidence of new infections of approximately 3 to 4 million
cases per year, HCV poses a major public health problem. More
efficacious and better-tolerated anti-HCV treatments are
therefore sorely needed to combat this major human pathogen.
Although the cloning of the HCV genome, twenty years ago,
allowed for a rapid analysis of the genomic organization and a
biochemical characterization of its proteins, the lack of a cell
culture system allowing efficient amplification of this virus has
long been a major obstacle for the study of its life cycle.
However, several tools have progressively been developed to
study this virus. The first major advance in the field has been the
development of a subgenomic replicon corresponding to a
replicative mini-genome, which was the first tool available to
study genomic replication. This discovery was followed by the
development of retroviral particles pseudotyped with HCV
envelope glycoproteins (HCVpp) to study HCV entry and to
38
Contents
the glycans associated with HCV envelope proteins. These
proteins are indeed heavily modified by conserved N-linked
glycans. By using site-directed mutagenesis, we have shown
that some glycans play a major role in the folding of E1E2
heterodimer or in virus entry. We are currently investigating the
role of the glycans associated with HCV envelope proteins in
the context of the HCVcc system. Furthermore, we have
demonstrated that other glycans present on E2 glycoprotein
contribute to the evasion of HCV from the humoral immune
response by masking the CD81 binding site. We have also
shown that the lectin cyanovirin-N inhibits HCV entry by
binding to HCV glycans and blocking the interaction between
the envelope protein E2 and CD81, indicating that HCV glycans
can be targeted for the development of new antiviral
approaches. We have also characterized the role of the
transmembrane domains of HCV envelope proteins in HCV
entry. Indeed, by using site directed mutagenesis and
tryptophan-replacement scanning of these regions, we have
demonstrated that the transmembrane domains of HCV
envelope glycoproteins play a major role in the fusion process.
Finally, with the use of HCVpp and HCVcc systems, we have
contributed to the characterization of human neutralizing
antibodies directed against the glycoprotein E2, and we have
shown that patients co-infected by HCV and HIV have lower
neutralizing antibody titers.
can affect HCV entry. We have shown that SAA, an acute phase
protein whose concentration increases markedly following
inflammation and infection, has antiviral activity against HCV.
In addition, the antiviral activity of SAA is strongly reduced in
the presence of HDL. These data indicate that HCV exploits HDL
and its receptor, SR-BI, to counteract the antiviral activity of SAA.
We are currently further investigating the mechanism of action
of SAA, as well as its in vivo relevance.
We have also investigated the role of plasma membrane lipids
in HCV entry. For this purpose, we first investigated the effect
of ceramide enrichment of the plasma membrane on HCV entry
by treating target cells with sphingomyelinase, which specifically catabolizes the sphingomyelin located on the outer leaflet
of the plasma membrane, leading to the formation of ceramide.
Our data show that sphingomyelinase inhibits HCV entry by
decreasing the cell surface expression of CD81. We are also
currently investigating the role of cholesterol in HCV entry.
Besides SR-BI, we have also analyzed the role of CD81 and some
of its partners in HCV entry. We have confirmed the role of CD81
in HCV entry within the framework of several collaborations.
Interestingly, members of the tetraspanin family are known to
interact with each other and with other transmembrane
proteins, thus building membrane multi-molecular complexes,
collectively referred to as the tetraspanin web. Importantly, we
have identified a new partner of CD81 which inhibits HCV entry
into Huh-7 cells by blocking E2-CD81 interaction. This CD81
partner, called EWI-2wint is a cleavage product of EWI-2, a major
partner of CD81, which belongs to a family of immunoglobulinlike proteins. Although EWI-2wint is expressed in different cell
lines, it is absent from hepatocytes, suggesting that, in addition
to the presence of specific entry factors in the hepatocytes, the
absence of a specific inhibitor may contribute to the
hepatotropism of HCV. We are currently characterizing the
mechanism of inhibition of HCV entry mediated by EWI-2wint.
Although the early steps of Hepatitis C Virus (HCV) entry into
the hepatocytes have yet to be elucidated, numerous studies
have shown a key role of the tetraspanin CD81 in this process.
In our lab, we have developed a human hepatoma Huh-7 cell
clone (Huh-7w7) having lost CD81 expression and that can be
infected by HCV when human CD81 is ectopically expressed.
Interestingly, we have shown that ectopic expression of mouse
CD81 (mCD81) in Huh-7w7 cells is also able to restore HCV
permissivity, indicating that mCD81 in the context of human
hepatocytes mimics human CD81 in HCV entry and likely in
interactions with cellular factors. We took advantage of these
permissive cells expressing mCD81 and the previously
described MT81/MT81w mAbs to analyze the role of
tetraspanin-enriched microdomain (TEM) associated CD81 in
HCV infection. These antibodies recognize murine CD81.
Importantly, the MT81w antibody, which only recognizes TEMassociated mCD81, does not inhibit HCV infection. Furthermore,
cholesterol depletion, which inhibits HCV infection and reduces
cell surface expression of total CD81, does not affect TEMassociated CD81 levels. Similarly, sphingomyelinase treatment,
which also reduces HCV infection and cell surface expression of
total CD81, raises TEM-associated CD81 levels. Altogether, our
data indicate that CD81 associated with TEM does not
participate in the early steps of the HCV life cycle.
After binding to specific receptor(s), virus entry into host cells
involves fusion of the lipid envelope with a cellular membrane.
This process is tightly coordinated in time and space and
requires drastic conformational changes in the fusion proteins,
which are triggered by cellular factors. Enveloped viruses enter
2. Cellular aspects of HCV entry
The recent development of functional models to analyze the
early steps of the HCV life cycle has led to the identification of
several cell surface proteins involved in HCV entry. The data that
have recently been accumulated suggest that HCV entry is a
slow and complex multi-step process. The exact role of each
molecule involved in HCV entry remains to be determined, but
current knowledge allows us to draw up a model. Glycosaminglycans and the LDL receptor may facilitate initial
attachment to the host cell. This interaction is probably
mediated by the lipoproteins associated with HCV virions.
However, one cannot exclude direct contact between HCV
envelope proteins and these cell surface molecules. After the
initial binding step, the particle likely interacts with the
scavenger receptor BI (SR-BI) and the tetraspanin CD81.
Although the sequence of HCV interaction with these two entry
factors has not been unequivocally determined, current
understanding suggests that a first contact with SR-BI might be
necessary before the particle interacts with CD81. The
interaction with SR-BI can potentially be direct or indirect
through HCV-associated lipoproteins. The tight-junction
molecule Claudin-1 (CLDN1) has also been identified as an
additional entry factor for HCV, which plays a role at a late stage
of the entry process, after interactions with SR-BI and CD81.
Since high-density lipoproteins (HDL) are the natural ligand of
SR-BI, we have tested the effect of these lipoproteins on HCV
entry. Interestingly, we have shown that HDL are able to
markedly enhance HCV entry. Furthermore, our data have
indicated a role for the lipid transfer activity of SR-BI in
facilitating HCV entry and suggest that HDL-mediated
enhancement of HCV entry involves a complex interplay
between SR-BI, HDL and HCV envelope glycoproteins.
Furthermore, we have also demonstrated that in addition to
increasing HCV entry, HDL also reduce the neutralizing activity
of anti-HCV antibodies. Based on these observations, we have
analyzed how another ligand of SR-BI, serum amyloid A (SAA),
39
Contents
for the assembly and release of HCV in cell culture and in vivo.
We have characterized the biogenesis of p7 and showed that
cleavages generating this polypeptide are inefficient and involve
specific structural determinants. We are currently trying to
understand the role of p7 in HCV morphogenesis by studying
its interaction with other proteins, its ion channel activity in
cellulo and its involvement in virus assembly with the HCVcc
system.
target cells by fusing their envelope with a cellular membrane.
This can occur at the plasma membrane or inside an endosome
after binding or after binding and endocytosis, respectively. In
this case, the acidic pH of endosomes triggers conformational
changes in the envelope proteins. With the use of endosome
acidification inhibitors, as well as siRNA or dominant-negative
proteins, we showed that HCV entry and that of the closely
related bovine viral diarrhea virus (BVDV), rely on clathrinmediated endocytosis and is pH dependent. For these studies,
we have benefited from the strong experience in cell biology
of one of our senior scientists, who has continued in parallel to
develop a project on the leptin receptor.
development 2010
With approximately 170 million infected people, hepatitis C virus (HCV)
is a major health problem worldwide. This virus often leads to chronic
infection that can progress towards chronic liver diseases, including
cirrhosis and hepatocellular carcinoma. The current antiviral therapy is
expensive, relatively toxic and effective in only half of treated patients,
and there is as yet no vaccine against this virus. Basic research on this
virus is therefore necessary for the development of new antiviral targets
to combat this major pathogen. The lack of a cell culture system
allowing efficient amplification of this virus has long been a major
obstacle for the study of its life cycle. However, the development of a
cell culture system that allows a relatively efficient amplification of HCV
(HCVcc) has finally recently been reported. HCVcc is now the most
relevant system to study the major steps of the HCV life cycle. For the
next 4 years, the activities of our team will therefore be focused on the
cell biology of the major steps of the HCV life cycle. We plan to continue
the characterization of HCV envelope glycoproteins and their
interaction with entry factors to better understand the mechanisms
leading to HCV entry. We will also study the interplay between
membrane trafficking and some steps of the viral life cycle and we will
investigate the mechanisms leading to HCV particle assembly.
3. HCV replication and virus assembly
With the recent development of a cell culture system for HCV,
we have also started to investigate some steps in virus assembly
and replication. The cell culture system for HCV allowed us for
the first time to characterize the subcellular localization of HCV
structural proteins in the context of an infectious cycle. Our data
have shown that, in infected cells, the glycoprotein heterodimer
is retained in the ER, and an association between the capsid
protein and lipid droplets was also observed. However, no clear
co-localization has been observed between the glycoprotein
heterodimer and the capsid protein in these infected cells.
Electron microscopy analyses allowed us to identify a
membrane alteration similar to the previously reported
"membranous web." However, no virus-like particles were found
in this type of structure. We have also tried to improve the
efficiency of replication of HCVcc by serial passages of the virus
in cell culture and by site-directed mutagenesis. Interestingly,
we have identified one major adaptive mutation located in the
E2 glycoprotein sequence. Furthermore, by site-directed
mutagenesis, we also showed that mutations of two major
positions coding for the C-terminal region of the core protein
could also increase the infectivity of HCVcc. This mutated virus
is more efficient for use as a tool in our studies. We have also
used this cell culture system to study the antiviral activity of
ribavirin. Our data suggest that the mutagenic effect of ribavirin
could in part explain its synergistic action with interferon.
Besides the structural proteins, some non-structural proteins can
also be involved in the assembly of HCV particle. One of these
proteins is the p7 polypeptide, which has been described as a
viroporin. Viroporins are small hydrophobic proteins that are
encoded in the genome of a wide variety of viruses and are
crucial for virion assembly and release. The p7 has been shown
to form ion channels in artificial membranes and to be crucial
Publications
2007
Antoine AF, Montpellier C, Cailliau K, Browaeys-Poly E, Vilain JP,
Dubuisson J.
The Alphavirus 6K protein activates endogenous ionic conductances
when expressed in Xenopus oocytes.
J Membr Biol, 2007, 215:37-48.
Akazawa D, Date T, Morikawa K, Murayama A, Miyamoto M,
Kaga M, Barth H,. Baumert T, Dubuisson J, Wakita T.
CD81 expression is important for heterogeneous HCV permissiveness of
Huh7 cell clones.
J Virol, 2007, 81, 5036-5045.
Brochot E, Duverlie G, Castelain S, Morel V, Wychowski C,
Dubuisson J, François C.
Effect of ribavirin on the hepatitis C virus (JFH-1) and its correlation with
interferon sensitivity.
Antivir Ther, 2007, 12:805-813.
Castelain S, Bonte D, Penin F, Francois C, Capron D, Dedeurwaerder S,
Zawadzki P, Morel V, Wychowski C, Duverlie G.
Hepatitis C Virus p7 membrane protein quasispecies variability in
chronically infected patients treated with interferon and ribavirin, with
or without amantadine.
J Med Virol, 2007, 79:144-154.
40
Contents
Dubuisson J, Helle F, Cocquerel L.
Early steps of the Hepatitis C Virus life cycle.
Cell Microbiol, 2008, 10:821-827.
Chapel C, Garcia C, Bartosh B, Roingeard P, Zitzmann N, Cosset FL,
Dubuisson J, Dwek RA, Trépo C, Zoulim F, Durantel D.
Reduction of the infectivity of hepatitis C virus pseudoparticles by
incorporation of misfolded glycoproteins induced by glucosidase inhibitors.
J Gen Virol, 2007, 88:1133-1143.
Helle F, Dubuisson J.
Hepatitis C Virus entry into host cells.
Cell Mol Life Sci, 2008, 65:100-112.
Chevalier C, Saulnier A, Benureau Y, Flechet D, Delgrange D,
Colbere-Garapin F, Wychowski C, Martin A.
Inhibition of Hepatitis C Virus Infection in Cell Culture by Small Interfering RNAs.
Mol Ther, 2007, 15:1452-1462.
Mechanism and inhibition of Hepatitis C Virus entry.
J Viral Entry, 2008, 3:61-67.
Ciczora Y, Callens N, Penin F, Pécheur EI, Dubuisson J.
The transmembrane domains of HCV envelope glycoproteins : residues
involved in E1E2 heterodimerization and involvement of these domains in
virus entry.
J Virol, 2007, 81:2372-2381.
Helle F, Cocquerel L.
L’entrée du virus de l’hépatite C dans ses cellules cibles.
Virologie, 2008, 12:105-116.
Jaffrelo L, Chabas S, Reigadas S, Pflieger A, Wychowski C, Rumi J,
Ventura M, Toulmé JJ, Staedel C.
A functional selection of viral genetic elements in cultured cells to identify
hepatitis C virus RNA translation inhibitors.
Nucleic Acids Res, 2008, 36:e95.
Couturier C, Sarkis C, Séron K, Belouzard S, Chen P, Lenain A, Corset L,
Dam J, Vauthier V, Dubart A, Mallet J, Froguel P, Rouillé Y, Jockers R.
Silencing of OB-RGRP in mouse hypothalamic arcuate nucleus increases
leptin receptor signaling and prevents diet-induced obesity.
Machida K, Kondo Y, Huang J, Chen YC, Cheng KTH, Keck Z,
Foung S, Dubuisson J, Sung VMH, Lai MMC.
HCV-induced Immunoglobulin Hypermutation Reduces the Affinity and
Neutralizing Activities of Antibodies against HCV Envelope Protein.
J Virol, 2008, 82:6711-6720.
Delgrange D, Pillez A, Castelain S, Cocquerel L, Rouillé Y,
Dubuisson J, Wakita T, Duverlie G, Wychowski C.
Robust production of infectious viral particles in Huh-7 cells by
introducing mutations in hepatitis C virus structural proteins .
Gen Virol, 2007, 88:2495-2503.
Molina S, Castet V, Pichard-Garcia L, Wychowski C, Meurs E,
Pascussi JM, Sureau C, Fabre JM, SaCunha A, Larrey D, Dubuisson J,
Coste J, McKeating J, Maurel P, Fournier-Wirth C.
Serum derived HCV infection of primary human hepatocytes is
tetraspanin CD81 dependent.
J Virol, 2008, 82:569-574.
Dubuisson J.
Hepatitis C virus proteins.
World J Gastroenterol, 2007, 13:2406-2415.
Duverlie G, Wychowski C.
Cell culture systems for the hepatitis C virus.
Rocha-Perugini V, Montpellier C, Delgrange D, Wychowski C,
Helle F, Pillez A, Drobecq H, Le Naour F, Charrin S, Levy S,
Rubinstein E, Dubuisson J, Cocquerel L.
The CD81 partner EWI-2wint inhibits hepatitis C virus entry.
PLoS ONE 3, 2008, e1866.
Fournier C, Duverlie G, Francois C, Schnuriger A, Dedeurwaerder S,
Brochot E, Capron D, Wychowski C, Thibault V, Castelain S.
A focus reduction neutralization assay for hepatitis C virus neutralizing
antibodies.
Virol J, 2007, 4:35.
Voisset C, Lavie M, Helle F, Op De Beeck A, Bilheu A, Bertrand-Michel J,
Tercé F, Cocquerel L, Wychowski C, Vu-Dac N, Dubuisson J. (2008)
Ceramide enrichment of the plasma membrane induces CD81
internalization and inhibits HCV entry.
Cell Microbiol, 2008, 10:606-617.
Gatignol A, Dubuisson J, Wainberg MA, Cohen EA, Jean-Luc Darlix JL.
New pandemics : HIV and AIDS, HCV and chronic hepatitis, Influenza virus
and flu.
Retrovirology, 2007, 4:8.
Voisset C, Weiss RA, Griffiths DJ.
Human RNA "rumor" viruses : the search for novel human retroviruses in
chronic disease.
Microbiol Mol Biol Rev, 2008, 72:157-196.
Helle F, Goffard A, Morel V, Duverlie G, McKeating J, Keck ZY,
Foung S, Penin F, Dubuisson J, Voisset C.
The neutralizing activity of anti-HCV antibodies is modulated by specific
glycans on the E2 envelope protein.
J Virol, 2007, 81:8101-8111.
2009
Johansson DX, Voisset C, Tarr AW, Aung M, Ball JK, Dubuison J,
Persson M.
Human combinatorial libraries yield rare antibodies that broadly
neutralize hepatitis C virus. Proc Natl
Acad Sci USA, 2007, 104:16269-16274.
Alvarez-Lajonchere L, Shoukry N, Gra B, Amado-Canizares Y, Helle F,
Bedard N, Guerra I, Drouin C, Dubuisson J, Gonzalez-Horta EE,
Martinez G, Marante J, Cinza Z, Castellanos M, Duenas-Carrera S.
Immunogenicity of CIGB-230, a therapeutic DNA vaccine preparation, in
HCV-chronically infected individuals in a phase I clinical trial.
J Viral Hepatitis, 2009, 16:156-167.
Lavie M, Goffard A, Dubuisson J.
Assembly of a functional HCV glycoprotein heterodimer.
Curr Issues Mol Biol, 2007, 9:71-86.
Charrin S, Yalaoui S, Bartosch B, Cocquerel L, Franetich JF, Boucheix
C, Mazier D, Rubinstein E, Silvie O.
The IG domain protein CD9P-1 down-regulates CD81 ability to support
plasmodium YOELII infection.
J Biol Chem, 2009, 284:31572-31578.
2008
Castelain S, Schnuriger A, François C, Nguyen-Khac E, Fournier C,
Schmit JL, Capron D, Dubuisson J, Wychowski C, Thibault V,
Duverlie G.
Lower levels of HCV neutralizing antibodies in HIV/HCV-coinfected
patients than in HCV-monoinfected patients.
J Infect Dis, 2008, 198:332-335.
Dubois A, Francois C, Descamps V, Fournier C, Wychowski C,
Dubuisson J, Castelain S, Duverlie G.
Enhanced anti-HCV activity of interferon alpha 17 subtype.
Virol J, 2009 , 6:70.
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Contents
Rôle des N-glycanes dans les fonctions des protéines d’enveloppe du virus
de l’hépatite C.
Virologie, 2009, 13 :271-282.
Montserret R, Saint N, Vanbelle C, Salvay A, Simorre JP, Ebel C,
Sapay N, Renisio JG, Böckmann A, Steinmann E, Pietschmann T,
Dubuisson J, Chipot C, Penin F.
NMR structure analysis and ion channel activity of the p7 polypeptide
from hepatitis C virus.
J Biol Chem, 2010, in press.
Kuzmina TI, Olenina LV, Sanzhakov MA, Farafonova TE,
Abramihina TV, Dubuisson J, Sobolev BN, Kolesanova EF.
Antigenicity and B-epitope mapping of hepatitis C virus envelope protein E2.
Biomed Chem (Moscow), 2009, 55:32-40.
Pawlotsky JM, Cocquerel L, Durantel D, Lavillette D, Lerat H,
Pécheur EI, Polyak SJ, Tautz N, Thimme R.
HCV research 20 years after discovery : a summary of the 16th
international meeting on hepatitis c virus and related viruses.
Rocha-Perugini V, Lavie M, Delgrange D, Canton J, Pillez A, Potel J,
Lecoeur C, Rubinstein E, Dubuisson J, Wychowski C, Cocquerel L.
The association of CD81 with tetraspanin-enriched microdomains is not
essential for Hepatitis C Virus entry.
BMC Microbiol, 2009, 9:111.
Popescu CI, Dubuisson J.
Role of lipid metabolism in hepatitis C virus assembly and entry.
Biol Cell, 2010, 102:63-74.
Roohvand F, Maillard P, Lavergne JP, Boulant S, Walic M, Andréo U,
Goueslain L, Helle F, McLauchlan J, Budkowska A.
Initiation of hepatitis C virus infection requires the dynamic microtubule
network: Role of the viral nucleocapsid protein.
J Biol Chem, 2009, 284:13778-13791.
Tews B, Popescu CI, Dubuisson J.
Last stop before exit - Hepatitis C assembly and release as antiviral drug targets.
Viruses, 2010, 2:1782-1803.
Vieyres G, Thomas X, Descamps V, Duverlie G, Patel AH, Dubuisson J.
Characterization of the envelope glycoproteins associated with infectious
hepatitis C virus.
J Virol, 2010, 84:10159-10168.
Tews BA, Cocquerel L.
Occludin, an additional key for hepatitis C virus entry.
Med Sci (Paris), 2009, 25:549-550.
Tews BA, Dubuisson J.
Occludin, the final essential factor for HCV entry ?
Future Virology, 2009, 4:329-333.
PhD
François HELLE
Directeur de thèse : J. Dubuisson
« les N-glycanes du Virus de l'Hépatite C protègent contre la neutralisation
mais sont une cible thérapeutique potentielle «
27 septembre 2007
Touvier T, Conte-Auriol F, Briand O, Cudejko C, Paumelle R, Caron S,
Baugé E, Rouillé Y, Salles JP, Staels B, Bailleul B.
LEPROT and LEPROTL1 cooperatively decrease hepatic growth hormone
action in mice.
J Clin Invest, 2009, 119 :3830-3838.
Vieyres G, Angus AG, Haberstroh A, Baumert TF, Dubuisson J, Patel AH.
Rapid synchronization of hepatitis C virus infection by magnetic adsorption.
J Virol Methods, 2009, 157:69-79.
David DELGRANGE
Directeur de thèse : C. Wychowski
« Etude de la multiplication de la souche JFH-1 du virus de l’hépatite C (VHC)
en cellules Huh-7. Adaptation et sélection de mutations permettant une
production rapide et massive du VHC. Localisation subcellulaire de
protéines du VHC. Mise en évidence de clones cellulaires résistants à
l’infection par le VHC »
21 septembre 2007
2010
Belouzard S.
Mécanismes d’entrée des coronavirus.
Virologie, 2010, 14:1-15.
Belouzard S, Madu I, Whittaker GR.
Elastase activation of the SARS coronavirus spike protein at discrete sites
within S2 domain.
J Biol Chem, 285:22758-22763.
Vera ROCHA
Directeur de thèse : L. Cocquerel
«Identification d’EWI-2wint, un partenaire de CD81 qui inhibie l’entrée du VHC»
26 septembre 2008
Ahmed-Belkacem A, Ahnou N, Barbotte L, Wychowski C, Pallier C,
Brillet R, Pohl RT, Pawlotsky JM.
Silibinin and related compounds are direct inhibitors of hepatitis C virus
RNA-dependent RNA polymerase.
HDR
Ngoc VU-DAC
« Rôle des lipoprotéines, transfert lipidique et mécanismes d’entrée du
VHC dans les hépatocytes »
2006
Ciczora I, Callens N, Séron K, Rouillé Y, Dubuisson J.
Identification of a dominant endoplasmic reticulum retention signal in
yellow fever virus pre-membrane protein.
J Gen Virol, 2010, 91:404-414.
Goueslain L, Alsaleh K, Horrelou P, Roingeard P, Descamps V,
Duverlie G, Ciczora Y, Wychowski C, Dubuisson J, and Rouillé Y.
Identification of GBF1 as a cellular factor required for HCV RNA replication.
J Virol, 2010, 84:773-787.
Laurence COQUEREL
« Identification et caractérisation d'un inhibiteur de l’entrée du virus de
l'hépatite C »
2006
Helle F, Vieyres G, Elkrief L, Popescu I, Wychowski C, Descamps V,
Castelain S, Duverlie G, Dubuisson J.
Role of N-linked glycans in the functions of HCV envelope proteins
incorporated into infectious virions.
J Virol, 2010, in press.
Karin SERON
« Etude de différents aspects du trafic membranaire : de la levure
Saccharomyces cerevisiae à l'obésité humaine »
2006
42
Contents
Paneth cells in mice. MDP is also a major component of
Freund's complete adjuvant that regulates antigen-specific
T-cell responses and antibody production. In collaboration
with Richard Flavell’s laboratory at the Yale University and
with Gabriel Nùñez at the University of Michigan, our data
revealed that MDP-triggered adaptive immunity is abolished
in Nod2-deficient animals. Further fundamental work is now
required to determine the cellular and molecular basis of the
immunomodulatory properties of MDP through NOD2mediated signalling. Consistent with our data, clinical studies
in Crohn’s Disease strongly support our hypothesis by
unravelling a deficiency in both alpha- and beta-defensins
and an impaired immunological response to MDP by beta
mononuclear cells isolated from peripheral blood.
NODS-Like Receptors in
26 Aug 2009
Mathias CHAMAILLARD, CR1 Inserm
Group Members :
Dessein Rodrigue, AHU Univ Lille 2/CHRU
Delacre Myriam, Technician IPL
Delanoye Anne, Technician IPL
Grandjean Teddy, Technician IPL
Normand Sylvain, Postdoctoral fellow IPL
Couturier Aurélie, PhD Student Inserm/NPdC
Key words :
Similarly to NOD2, we identified that NOD1-dependent
signalling is activated in response to the dipeptide
γ-D-glutamyl-meso-diaminopimelic acid (iE-DAP) or γ-Dglutamic-meso-diaminopimelic acid (iQ-DAP). iE-DAP and
iQ-DAP represent a signature of bacterial infection in that
such motifs are not expressed by eukaryotic cells.
Consistently, our data revealed an essential role of NOD1 in
limiting infection by both Gram-negative and Gram-positive
bacteria in vivo. By using a biochemical and systematic
mutational analysis, we identified a general mechanism by
which NOD1 and NOD2 detect different structures of the PGN
through their leucine-rich repeats. The N-terminal and
C-terminal leucine-rich repeats were differentially required in
the engagement of NOD2-mediated signalling by MDP.
Notably, the residues thought to be involved in the
βstrand/βturn of the C-terminal leucine-rich repeats are
essential for the MDP-induced NF-κB activation. We also
identified specific residues and essential regulatory domains
of NOD1 and NOD2 involved in NF-κB activation. Lastly, our
data provided a better understanding of the molecular basis
of disease susceptibility by revealing that loss-of-function
and gain-of-function mutations are predisposing respectively
to Crohn’s disease and to Blau syndrome, another
granulomatous disorder that affects solely aseptic sites.
Inflammatory bowel diseases. Infection. Inflammation. Innate
immunity. Intestinal homeostasis. Microbiota. Mucosal immunity. Systemic immunity. Tolerance. Tumorigenesis.
A - Research Report
Rationale. The gastrointestinal mucosa enables tolerance
towards commensals whereas it promotes a robust
antimicrobial response to enteropathogens when they first
breach the integrity of the mucosal barrier. While food-borne
pathogens remain a major cause of morbidity and mortality
worldwide, the intestinal microbiota is also involved in the
pathogenesis of several incurable and unpredictive chronic
inflammatory diseases, including Crohn’s disease, colorectal
cancer and asthma. The molecules involved in this dynamic
surveillance machinery still remain an unresolved theme in
medicine and biology.
Main Results. Recent evidence, including work from us,
revealed an essential role of NOD2 in anti-bacterial immunity
and in the pathogenesis of Crohn’s Disease, a prototypical
and incurable form of inflammatory bowel disease that
affects millions worldwide. NOD2 is a member of the recently
identified family of cytosolic NODs-Like Receptors (NLRs),
which share similarities with the superfamily of plant diseaseresistance proteins. Similarly to the membrane-bound
Toll-like receptors, certain NLRs play an essential role in
detecting a diversified set of threats that primarily originate
from microorganisms and cell remnants. NLRs-mediated
signalling is thought to recruit and activate downstream
effectors such as mitogen-activated protein kinase (MAPK)
and transcription factors, including nuclear factor-kappa B.
Among the elicitors that are also thought to play a role in
innate immunity and in the pathogenesis of chronic
inflammatory bowel diseases, we have then focused our
interest on the physiological role of the soluble c-type lectin
REG3β, in that the mechanisms whereby REG3β is involved in
intestinal homeostasis remain elusive. Our most recent data
revealed a new regulatory function in intestinal inflammation
and host defence for REG3β. While REG3β expression is
induced during both colitis and enteric infections, its absence
limits intestinal injury and bacterial clearance. Release of
REG3β is coordinated by the TLR2-mediated recognition of
components of the microbiota independently of NOD2.
Treatment with exogenous REG3β significantly triggered
neutrophil recruitment through IL-17A and secretion of
CXCL1 production in vivo. Consistently, DSS-induced injury
triggered impaired neutrophil influx and reduced signs in
REG3β−/− mice when compared to their wild-type
littermates. In the absence of REG3β, the composition of the
microbiota was profoundly altered with an increased fecal
prevelance of Gram-positive bacteria. Colonic REG3β
expression was strongly correlated to transcript levels of IL-8
in CD patients. Collectively, we have identified that REG3β is
involved in neutrophil recruitment through IL-17A and has a
Our recent studies have focused on the respective roles of
NOD2 and NOD1, the NLR molecule closest to NOD2. To gain
further insights into the role of NOD2, we first identified
MurNAc-L-Ala-D-isoGln (MDP) as the essential microbial
structure sensed by NOD2 ex vivo and in vivo. MDP is a
structure commonly found in peptidoglycan from both
Gram-positive and Gram-negative bacteria. The absence of
NOD2 confers increased predisposition to oral, but not
systemic, infection by a restricted panel of infectious agents,
including Listeria monocytogenes but not Yersinia
pseudotuberculosis. Importantly, we found that NOD2
regulates the expression of alpha-defensins produced by
43
Contents
Approaches : We intend to work out the physiological role of
NOD2 from the whole organism to the molecular level.
pathogenic role in gut inflammation, supporting the
development of therapeutic approaches targeting REG3β to
treat relapsing-remitting inflammation in these patients.
The first theme is to determine its biological effect on the
intestinal microbiota and which specific components of
commensals may affect engagement of NOD2-mediated
signaling pathway. To achieve this first specific aim, we will
therefore combine systematic molecular examination of the
gut microbial community and macroarray gene expression
analysis on micro-dissected intestinal areas (i.e. epithelial vs
lamina propria) of antibiotic-treated mice and (monocolonized) germ-free animals.
In parallel, we recently demonstrated that the nuclear receptor
peroxisome proliferator-activated receptor-gamma (PPARγ) is
essential for intestinal homeostasis in response to both
dietary- and microbiota-derived signals. Its role in host
defense remains unknown, however. We showed that PPARγ
functions as an antimicrobial factor by maintaining
constitutive epithelial expression of a subset of β-defensin in
the colon, which include mDefB10 in mice and DEFB1 in
humans. Colonic mucosa of Pparγ-mutant animals shows
defective killing of several major components of the intestinal
microbiota which include Candida albicans, Bacteroides
fragilis, Enterococcus faecalis and Escherichia coli.
Neutralization of the colicidal activity by using an antimDefB10 blocking antibody was effective in a
PPARγ-dependent manner. A functional promoter variant that
is required for DEFB1 expression confers strong protection
against Crohn’s colitis and ileocolitis (p=0.018; OR=0.559).
Consistently, colonic involvement in CD is specifically linked
to reduced expression of DEFB1 independently of
inflammation. These findings support the development of
PPARγ-targeting therapeutic and/or nutritional approaches to
prevent colonic inflammation by restoring antimicrobial
immunity in CD.
The second theme aims at elucidating the critical regulatory
processes involved in the engagement of NOD2 activation in
intestinal homeostasis, we will therefore determine its
respective role within cells that are patrolling or lining the
intestinal mucosa. To achieve this result, the project combines
the use of knockout mice models with systematic molecular
analysis of the microbiota, gene expression on microdissected
intestine, in vivo imaging analysis, bone marrow transfer
experiments and relevant cell-based assays.
Originality : Our global approach will improve our
understanding of intestinal tolerance and of immunoregulatory properties of the microbiota that constitute a major
unresolved theme in biology and medicine. Our original
proposal might therefore have an impact on disease
pathogenesis and on the worldwide biomedical research by
manipulating NOD2 function in vivo for preventing these
widespread illnesses.
development 2010
Publications
Research focus : Despite significant progress in the field, the
characterization of regulators of innate immunity in the
dialogue between the host and the microbiota is currently at
the fundamental research stage. Several challenging issues
still remain unresolved, however. Notably, it remains to be
seen what is the impact(s) of specific components of the
microbiota in host defence, tumorigenesis and autoimmunity
through NOD2 ? Which are the regulatory mechanisms
accounting for the tissue-restricted phenotype of defective
NOD2 signalling, and particularly so at the level of cells that
are lining or patrolling the interface with the luminal
digestive environment? In this context, the scientific
objectives of our junior research group aims now at
deciphering the role of NOD2 in the gut-microbiota dialogue
and at dissecting its sensitizing or protecting impact on the
prevalence and severity of chronic inflammatory diseases at
the cellular level within the gut mucosa.
2007
Chakass D, Philippe D, Erdual E, Dharancy S, Malapel M,
Dubuquoy C, Thuru X, Gay J, Gaveriaux-Ruff C, Dubus P,
Mathurin P, Kieffer BL, Desreumaux P, Chamaillard M.
mu-Opioid receptor activation prevents acute hepatic inflammation and
cell death.
Gut, 2007, 56:974-981.
Foligné B, Dessein R, Marceau M, Poiret S, Chamaillard M, Pot B,
Peyrin-Biroulet L, Chamaillard M.
NOD2 and defensins : translating innate to adaptive immunity in Crohn's
disease.
J endotoxin Res, 2007, 13:135-139.
Working hypothesis : The overall hypothesis guiding the
objectives of the research is that impaired innate immune
surveillance of the microbiota at the intestinal barrier level
may hasten clinical symptoms through the host’s failure to
protect against tissue damage and to drive an optimal
immunological response. If this is true, we should be able to
identify rational and novel targets with a view to curing
inflammatory barrier diseases and/or preventing their natural
progression. To this end, several non-exclusive
physiopathological hypotheses are tested.
Peyrin-Biroulet L, Chamaillard M, Gonzalez F, Beclin E, Decourcelle
C, Antunes L, Gay J, Neut C, Colombel JF, Desreumaux P.
Mesenteric fat in Crohn's disease : A pathogenetic hallmark or an innocent
bystander ?
Gut, 2007, 56:577-583.
Peyrin-Biroulet L, Rodriguez-Gueant RM, Chamaillard M,
Desreumaux P, Xia B, Bronowicki JP, Bigard MA, Gueant JL.
Vascular and cellular stress in inflammatory bowel disease : Revisiting the
role of homocysteine.
Am J Gastroenterology, 2007, 102:1108-1115.
44
Contents
Koslowski MJ, Kübler I, Chamaillard M, Schaeffeler E, Reinisch W,
Wang G, Beisner J, Teml A, Peyrin-Biroulet L, Winter S, Herrlinger KR,
Rutgeerts P, Vermeire S, Cooney R, Fellermann K, Jewell D,
Bevins CL, Schwab M, Stange EF, Wehkamp J.
Genetic variants of Wnt transcription factor TCF-4 (TCF7L2) putative
promoter region are associated with small intestinal Crohn's disease.
PLoS One, 2009, 4:e4496.
Rousseaux C, Thuru X, Gelot A, Barnich N, Neut C, Dubuquoy L,
Dubuquoy C, Merour E, Geboes K, Chamaillard M. Ouwehand A,
Leyer G, Carcano D, Colombel JF, Ardid D, Desreumaux P.
Lactobacillus acidophilus modulates intestinal pain and induces opioid
and cannabinoid receptors.
Nat Med, 2007, 13:35-37.
Nod-like receptors: Cytosolic watchdogs for immunity against pathogens.
Plos Pathogens, 2007, 3:e152.
Standaert-Vitse A, Sendid B, Joossens M, François N, Vandewalle-El
Khoury P, Branche J, Van Kruiningen H, Jouault T, Rutgeerts P,
Gower-Rousseau C, Libersa C, Neut C, Broly F, Chamaillard M,
Vermeire S, Poulain D, Colombel JF.
Candida albicans colonization and ASCA in familial Crohn's disease.
Am J Gastroenterol, 2009, 104:1745-1753.
Vignal C, Singer E, Peyrin-Biroulet L, Desreumaux P, Chamaillard M.
How NOD2 mutations predispose to Crohn's disease ?
Chamaillard M.
NLRs : a Cytosolic Armory of Microbial Sensors Linked to Human Diseases.
Book chapter in Nucleic Acids and Molecular Biology, 2007.
2010
Joossens M, Van Steen K, Branche J, Sendid B, Rutgeerts P,
Vasseur F, Poulain D, Broly F, Colombel JF, Vermeire S, Chamaillard M.
Familial aggregation and antimicrobial response dose-dependently
affect the risk for Crohn's disease.
Inflamm Bowel Dis, 2010, 16:58-67.
2008
Body-Malapel M, Dharancy S, Berrebi D, Louvet A, Hugot JP,
Philpott DJ, Giovannini M, Chareyre F, Pages G, Gantier E,
Girardin SE, Garcia I, Hudault S, Conti F, Sansonetti PJ,
Chamaillard M, Desreumaux P, Dubuquoy L, Mathurin P.
NOD2 : a potential target for regulating liver injury.
Lab Invest, 2008, 88:318-327.
Peyrin-Biroulet L, Beisner J, Wang G, Nuding S, Oommen ST, Kelly D,
Parmentier-Decrucq E, Dessein R, Merour E, Chavatte P, Grandjean
T, Bressenot A, Desreumaux P, Colombel JF, Desvergne B, Stange EF,
Wehkamp J, Chamaillard M.
Peroxisome proliferator-activated receptor gamma activation is required
for maintenance of innate antimicrobial immunity in the colon.
Proc Natl Acad Sci U S A, 2010, 107:8772-8777.
Dessein R, Chamaillard M, Danese S.
Innate immunity in Crohn's disease: the reverse side of the medal.
J Clin Gastroenterol, 2008, 42 Suppl 3 Pt 1:5144-5147.
Rehaume LM, Jouault T, Chamaillard M.
Lessons from the inflammasome : a molecular sentry linking Candida and
Crohn's disease.
Trends Immunol, 2010, 31:171-175.
Vandewalle-El Khoury P, Colombel JF, Joossens S, Standaert-Vitse A,
Collot M, Halfvarson J, Ayadi A, Landers CJ, Vermeire S, Rutgeerts P,
Targan SR, Chamaillard M, Mallet JM, Sendid B, Poulain D.
Detection of antisynthetic mannoside antibodies (A Sigma MA) reveals
heterogeneity in the ASCA response of Crohn's disease patients and
contributes to differential diagnosis, stratification, and prediction.
Secher T, Gaillot O, Ryffel B, Chamaillard M.
Remote control of intestinal tumorigenesis by innate immunity.
Cancer Res, 2010, 70:1749-1752.
Wolowczuk I, Allez M, Chamaillard M.
IL-17/23, potential targets for Crohn’s disease.
Book chapter in series Progress in Inflammation Research, 2008.
PhD
Branche J, Chamaillard M, Colombel JF.
Antibodies : useful tools or pathophysiology markers ?
Book chapter in Falk Symposium 2008.
Rodrigue DESSEIN
Directeur de thèse : Michel Simonet
ou
Co-Directeur : Mathias Chamaillard
Sujet de thèse : Réponse immunitaire de la muqueuse intestinale à
l’agression par Yersinia pseudotuberculosis
29 septembre 2008
2009
André MF, Aumaître O, Grateau G, Chamaillard M, CostedoatChalumeau N, Cardoso MC, Henry-Berger J, Ramakrishna BS,
Delpech M, Piette JC, Creveaux I.
Longest Form of CCTG Microsatellite Repeat in the Promoter of the
CD2BP1/PSTPIP1 Gene Is Associated with Aseptic Abscesses and with
Crohn Disease in French Patients.
Dig Dis Sci, 2009, Sep 3. [Epub ahead of print].
HDR
Mathias CHAMAILLARD
Diplôme : Habilitations à diriger des recherches
18 décembre 2008
Dessein R, Peyrin-Biroulet L, Chamaillard M.
Intestinal microbiota gives a nod to the hygiene hypothesis in type 1
diabetes.
Secher T, Lacas-Gervais S, Gratadoux JJ, Lafont F. Dagorn JC,
Ryffel B, Akira S, Langella P, Nùñez G, Sirard JC, Iovanna J,
Simonet M, Chamaillard M.
Toll-like receptor 2 is critical for induction of Reg3 beta expression
and intestinal clearance of Yersinia pseudotuberculosis.
Gut, 2009, 58:771-776.
45
Contents
1. Anti-Microbial Mucosal Immunity
Lung Infection
and Innate Immunity
(J.C. Sirard - C. Carnoy, U801)
This group focused on the early mechanisms leading to
mucosal immunity during bacterial infection and on the
contribution of virulence factors in the pathogenesis.
Flagellin, a conserved motif expressed by flagellated bacteria,
was used as a model to dissect the mechanisms leading to
lung immunity. Our data showed that intranasal instillation of
flagellin stimulates the pulmonary expression of various genes
including CCL20, a chemokine involved in DC recruitment.
Using whole lung transcriptome and tissue microdissection,
we found that airway epithelial cells (AEC) mediate the
response. We have also studied the role of flagellin domains
on the TLR5-mediated immune response. Interestingly, whilst
the conserved domain of flagellin is pivotal to initiate mucosal
and systemic immunity, the hypervariable region contributes
to trigger systemic pro-inflammatory response.This finding
might have implications in better exploiting the adjuvant
property of flagellin.
In parallel, this group has investigated the role of the
superantigen YPM from the enterobacterium Yersinia
pseudotuberculosis in immune response. The data show that,
in contrast to other superantigens, the immunopathological
effect of YPM is mediated by cellular toxicity in infected hosts.
François TROTTEIN, DR2 CNRS
Group Members :
Carnoy Christophe, MCU Univ Lille 2
Faure Karine, MCU-PH Univ Lille 2
Faveeuw Christelle, CR1 Inserm
Gosset Philippe, CR1 Inserm
Guery Benoît, PUPH Univ Lille2/CHRU
Kipnis Eric, PHU Univ Lille 2
Pichavant Muriel, CR2 Inserm
Sirard Jean-Claude, CR1 Inserm
Tillie-Leblond Isabelle, Doct Med PH Univ Lille 2/CHRU
Fontaine Josette, IE1 IPL
Tabareau Julien, IE IPL (CDD)
Cayet Delphine, Technician IPL
Vendeville Catherine, Technician IPL
Vilain Eva, Technician IPL
Paget Christophe, Postdoctoral fellow
Renneson Joëlle, Postdoctoral fellow
Bialecki Emilie, PhD Student MENRT
Ivanov Stoyan, PhD Student MENRT
Macho-Fernandez Elodie, PhD Student Univ Lille 2
Van Maele Laurye, PhD Student Univ Lille 2
Fougeron Delphine, M2 Pro Université de Tours
Mear Jean Baptiste, Master 2 recherche, Univ de la Méditerranée
Rouyer Cécile, Master 2 recherche, Univ de la Méditerranée
2. Respiratory and Circulatory Distress
(B. Guery, K. Faure, E. Kipnis, EA 2689)
Key words :
This group mainly focused on Pseudomonas aeruginosa
pneumonia. P. aeruginosa is a Gram negative bacterium that
produces a wide spectrum of virulence factors. Among them,
the type III secretion system (TTSS) promotes the cytosolic
injection of 4 exotoxins into target cells. Our results show that
TTSS is a key virulence determinant in vivo and represents a
major target for therapeutic purposes. The potential
protective effects of humanized anti-TTSS antibodies are
presently tested in patients. In P. aeruginosa, the production
of certain virulence factors is regulated by quorum sensing
(QS), a bacterial communication system allowing bacterial
populations to coordinate their responses. Using clinical
samples (ventilator-associated pneumonia), we observed a
correlation between the level of QS activity and the
production of virulence factors, particularly bacterial elastase.
We have also successfully tested the clinical efficacy of
antibiotics in cystic fibrosis patients to modulate the
production of QS-dependent virulence factors. Finally, we
found that modulation of host factors, such as coagulation
and apoptosis, as well as polyunsaturated fatty acid balance
influence P. aeruginosa-mediated pathology in mice.
Respiratory pathogens, Pulmonary inflammation, Innate
immunity, Airway epithelium, Dendritic cells, NKT cells.
A - Research Report
The team “Lung Infection and Innate Immunity” (LIII) was
created in January 2010 and originates from four research
groups having a long-standing interest in innate immune
responses, (pulmonary) inflammation and/or infectious
diseases.
* The group “Anti-Microbial Mucosal Immunity” is headed by
Dr Jean-Claude SIRARD and belonged to Inserm U801
“Cellular and Molecular Interactions of Bacterial Pathogens
with the Host: the Yersinia Model” (Director: Prof Michel
Simonet).
* The group «Laboratoire de Recherche en Pathologie
Infectieuse» was part of EA 2689, University of Lille 2
«Détresses Respiratoires et Circulatoires” (Director: Prof Regis
Matran) and was headed by Prof. Benoit GUERY.
* The group «Epithelial interface in pulmonary inflammation»
is headed by Dr Philippe GOSSET and belonged to Inserm
U774 “Biomolecules and Pulmonary Inflammation” (Director:
Dr Philippe Lassalle).
* The group “Innate Immunity and Immunoregulation” is
headed by Dr François TROTTEIN and belonged to Inserm
U547, “Schistosomiase, Paludisme et Inflammation” (Director:
Prof Monique Capron).
3. Epithelial interface in pulmonary
inflammation (P. Gosset - I. Tillie-Leblond, U774)
This group aimed at better understanding the role of airway
epithelium in allergic asthma. The mechanisms leading to
airway remodeling, in particular in children developing severe
asthma, have been examined. Expression of some matrix
metalloproteinases (MMP) in pulmonary tissues (AEC)
correlates with the intensity of allergic reaction. In the mouse
system, ADAM-12 MMP plays key role in asthma by enhancing
46
Contents
the mobilization of inflammatory cells through CXCL1 and
CXCL8. Airway epithelium shedding and increased lung
permeability are characteristics of severe asthma. We showed
that the local delivery of the epithelial growth factor KGF is
beneficial in the rat system. Together, our data suggest the
potential clinical interest of MMPs and KGF in severe asthma.
In parallel, this group has investigated the role of innate
sensors expressed by airway epithelium in asthma
development. Our finding revealed that AEC sense allergens,
as well as microbial components, to mobilize dendritic cells
into the lungs and to trigger lung immunity. In this setting, we
have highlighted the role of some scavenger receptors
(CXCL16 and LOX-1) and TLR members. More recently, the role
of TLR3 expressed by AEC in the recognition of viral
component (dsRNA) has been underlined, a finding that may
be relevant to better understand the mechanisms leading to
virus-triggered asthma exacerbation.
factors in the pathogenesis. We will examine this issue using
Streptococcus pneumoniae, known to develop strategies to
colonize the respiratory tract and to invade the
reticuloendothelial system, thereby causing bacteremia. To
this end, we will compare the early immune response and
developed disease in resistant vs susceptible mice infected
with fully virulent bacteria or virulence-attenuated mutants.
2. Respiratory infection and chronic
airway inflammation (P. Gosset)
This theme is intended to determine the role of respiratory
pathogens in the development of asthma and COPD. It is well
established that respiratory infections and chronic lung
inflammatory diseases impact on each other. Respiratory
viruses are associated with the initiation and/or exacerbation
of allergic asthma, notably during childhood. COPD patients
are highly susceptible to pulmonary bacterial infections which
in turn exacerbate the pathogenesis of COPD. We aim at
defining the molecular and cellular mechanisms involved in
asthma initiation by influenza A virus (IAV) and COPD
exacerbation by S. pneumoniae. A particular focus will be
placed on the role of AEC/innate immune cell interplays in
host defence and pathology. Experimental mouse models as
well as clinical samples will be used to address this question.
4. Innate Immunity and Immunomodulation (F. Trottein - C. Faveeuw, U547)
The main interest of this group is to better understand innate
mechanisms leading to T helper polarization during infection.
In this setting, through their ability to explosively secrete
immunoregulatory cytokines, Natural Killer T (NKT) cells play
pivotal functions, although their mode of activation is still
elusive. NKT cells recognize lipid Ags presented by the MHC
class I-like protein CD1d expressed by APC and play part in a
broad range of pathologies. Using a model of helminth
infection (Schistosoma mansoni), this group showed that NKT
cells play pivotal functions on the Th2 response and
pathology. This group has also recently identified new modes
of NKT cell activation in response to parasite (S. mansoni) and
viral stimuli. In these settings, some host factors (CD1drestricted lipids, cytokines) contribute to NKT cell activation.
Finally, this group has shown that some, but not all APC
(sub)populations are important to activate and polarize NKT
cells in vivo, a finding that might also ultimately lead to better
control these cells for therapeutical purposes.
3. NKT cells during respiratory
infection and chronic airway
inflammation (F. Trottein)
NKT cells are important in host defence and pathology during
infection but the initial trigger and the mechanisms
underlying their activation, as well as their precise functions
during respiratory infection, are ill-defined. The objective of
this theme is to examine these questions in different
experimental models of viral (IAV) and bacterial
(S. pneumoniae) (co)-infection. Their role in asthma and COPD
exacerbation by viral and bacterial infection will be also
examined. We also aim at better manipulating these cells for
therapeutic purposes.
development 2010
4. Conclusions
Our proposal should (1) provide an insight into the nature of
the early host factors influencing innate immune and (chronic)
inflammatory responses during pulmonary bacterial and viral
infections and (2) lead to the identification and exploitation
of novel immunomodulatory components and of new cellular
and molecular targets for the development of improved
therapies and vaccines.
The objective of the team “Lung Infection and Innate
Immunity” is to investigate the role and the mode of activation
of innate immune cells, either as sentinels or effectors, in host
defence and pulmonary pathogenesis during respiratory
bacterial and viral infections. Experimental mouse models as
well as clinical studies will be used to address this question.
Three main themes will be developed.
1. Respiratory bacterial infection and
innate immunity (JC. Sirard)
The objective of this theme is to define innate defence
mechanisms involved in the early clearance of respiratory
bacterial pathogens and to determine the role of virulence
47
Contents
Michel ML, Castro-Keller A, Paget C, Trottein F, Schneider E, Dy M,
Leite de Moraes M.
Identification of a new IL-17-producing iNKT cell subset involved in airway
neutrophilia.
J Exp Med, 2007, 204:995-1001.
Publications
2007
Ader F, Le Berre R, Lancel S, Faure K, Viget NB, Nowak E, Neviere R,
Guery BP.
Inhaled nitric oxide increases endothelial permeability in Pseudomonas
aeruginosa pneumonia.
Intensive Care Med, 2007, 33:503-510.
Molle C, Nguyen M, Flamand V, Trottein F, Willems F, Goldman M
and Goriely S.
IL-27 Synthesis Induced by TLR Ligation Critically Depends on IFN
Regulatory Factor 3.
J Immunol, 2007, 178:7607-7615.
Breuilh L, Vanhoutte F, Fontaine J, van Stijn CM, Tillie-Leblond I,
Capron M, Faveeuw C, Jouault T, van Die I, Gosset P, Trottein F.
Galectin-3 modulates immune and inflammatory responses during
helminthic infection: impact of galectin-3 deficiency on the functions of
dendritic cells.
Infect Immun, 2007, 75:5148-5157.
Paget C, Mallevaey T, Speak AO, Torres D, Fontaine J, Sheehan KC,
Capron M, Ryffel B, Faveeuw F, Leite de Moraes M, Platt F, Trottein F.
Activation of iNKT cells by TLR9-stimulated myeloid dendritic cells requires
type I interferon and charged glycosphingolipids.
Immunity, 2007, 27:597-609.
Pierre M, Husson MO, Leberre R, Desseyn JL, Galabert C, Beghin L,
Beermann C, Dagenais A, Berthiaume Y, Cardinaud B, Barbry P,
Gottrand F, Guery BP.
Omega 3 polyunsaturated fatty acids improve host response in chronic
Pseudomonas aeruginosa lung infection in mice.
Am J Physiol Lung Cell Mol Physiol, 2007, 292:1422-1431.
Foligne B, Zoumpopoulou G, Dewulf J, Chareyre F, Sirard JC, Pot B,
Grangette. C.
A key role of dendritic cells in probiotic functionality.
PLoS ONE, 2007, 2:e313.
Gosset P, Tillie-Leblond I, Le Berre R, Janin A, Prangère T, Tonnel AB,
Guery BP.
KGF improves lung permeability increase and epithelium alterations in
allergic rats.
Eur Respir J, 2007, 30:31-39.
Nod-like receptors : cytosolic watchdogs for immunity against pathogens.
PLoS Pathog, 2007, 3:e152.
Speak A, Neville D, Priestman D, Salio M, Platt N, Heare T,
Butters TD, Dwek RA, Trottein F, Exley M, Cerundolo V, Platt F.
Restricted presence of isoglobotrihexosylceramide in mammals :
implications for the identity of NKT cell ligands.
Guo A, M. Lasaro A, Sirard JC, Kraehenbuhl JP, Schifferli DM.
Adhesin-dependent binding and uptake of Salmonella enterica
serovar Typhimurium by dendritic cells.
Microbiology, 2007, 153:1059-1069.
Hammad A, Kool M, Soullié T, Narumiya S, Trottein F, Hoogsteden
Lambrecht B.
Activation of the D prostanoid 1 receptor suppresses asthma by
modulation of lung dendritic cell function and induction of regulatory
cells.
J Exp Med, 2007, 204: 357-367.
Tetaert D, Pierre M, Demeyer D, Husson MO, Béghin L, Galabert C,
Gottrand F, Beermann C, Guery BP, Desseyn JL.
Dietary n-3 fatty acids have suppressive effects on mucin upregulation
in mice infected with Pseudomonas aeruginosa. Respir Res, 2007, 5:39.
Tillie-Leblond I, Gosset P, Le Berre R, Janin A, Prangère T, Tonnel AB,
Guery BP.
Keratinocyte growth factor improves alterations of lung permeability and
bronchial epithelium in allergic rats.
Eur Respir J, 2007, 30:31-39.
Hatzfeld-Charbonnier AS, Lasek A, Gosset P, Velu T, Formstecher P,
Mortier L, Marchetti P.
Influence of heat stress on human monocyte-derived dendritic cell
functions with immunotherapeutic potential for antitumor vaccines.
J, Leukoc, Biol, 81:1179-1187.
Vanhoutte F, Breuilh L, Fontaine J, Zouain C, Mallevaey T, Vasseur V,
Capron M, Goriely S, Faveeuw F, Ryffel B, Trottein F.
Toll-like receptor (TLR)2 and TLR3 sensing is required for dendritic cell
activation, but dispensable to control Schistosoma mansoni infection
and pathology.
Just N, Moreau C, Lassalle P, Gosset P, Perez T, Wallaert B, Destee A,
Defebvre L, Tonnel AB, Devos D.
High erythropoietin and low vascular endothelial growth factor levels in
cerebrospinal fluid from hypoxemic ALS patients suggest an abnormal
response to hypoxia.
Neuromuscul. Disord, 2007, 17:169-173.
Wesley JD, Tessmer MS, Paget C, Trottein F and Brossay L.
A Y-chromosome linked factor impairs iNK T development.
J Immunol, 2007, 179:3480-3487.
Kipnis E, Guery BP.
Targeting lung coagulation disorders in acute lung injury : easy as pie
(PAI-1) ?
Crit Care Med, 2007, 35:1448-1449.
2008
Kipnis E, Guery BP, Wiener-Kronish J.
Promises and limitations of the bronchoscopic microsampling probe.
Chest, 2007, 132:1414.
Ader F, Faure K, Guery BP, Nseir S.
Pseudomonas aeruginosa and Candida albicans interaction in the
respiratory tract: From pathophysiology to a therapeutic perspective.
Pathol Biol (Paris), 2008, 56:164.
Mallevaey T, Fontaine J, Breuilh L, Paget C, Vendeville C, CastroKeller A, Capron M, Leite de Moraes M, Trottein F, Faveeuw C.
Invariant and non-invariant Natural Killer T cells exert opposite regulatory
function the immune response during murine schistosomiasis.
Infect Immun, 2007, 75:2171-2180.
Ader F, Le Berre R, Fackeure R, Raze D, Menozzi FD, Viget N, Faure K,
Kipnis E, Guery BP, Jarraud S, Etienne J, Chidiac C.
In vivo effect of adhesion inhibitor heparin on Legionella pneumophila
pathogenesis in a murine pneumonia model.
Intensive Care Med, 2008, Mar 26.
48
Contents
Depontieu F, Grigoriu BD, Scherpereel A, Adam E, Delehedde M,
Gosset P, Lassalle P.
Loss of Endocan tumorigenic properties after alternative splicing
of exon 2.
BMC Cancer, 2008, 8:14.
Torres D, Paget C, Fontaine J, Mallevaey T, Matsuoka T, Maruyama
T, Narumiya S, Capron M, Gosset P, Faveeuw C, and Trottein F.
Prostaglandin D2 Inhibits the Production of IFN-{gamma} by Invariant
NK T Cells : Consequences in the Control of B16 Melanoma.
J Immunol, 2008, 180:783.
Didierlaurent A, Goulding J, Patel S, Snelgrove R, Low L, Bebien M,
Lawrence T, van Rijt LS, Lambrecht BN, Sirard JC, Hussell T.
Sustained desensitization to bacterial Toll-like receptor ligands after
resolution of respiratory influenza infection.
J Exp Med, 2008, 205:323.
Vanhoutte F, Breuilh L, Paget C, Fontaine J, Goriely S, Ryffel B and
Trottein F.
Toll-like receptor (TLR)2 and TLR3 synergy and cross-inhibition in murine
myeloid dendritic cells.
Immunol Lett, 2008, 116:86.
Faveeuw, C., Mallevaey, T, and Trottein, F.
Role of Natural Killer T lymphocytes during helminth infections.
Parasite, 2008, 15:284-288.
2009
Betbeder D, Trottein F.
The effect of alpha-galactosylceramide on the activation of Natural Killer
T lymphocytes.
Vaccine, 2009, 27:3691.
Guery BP, Guidet B, Beloucif S, Floret D, Le Gall C, Chouaid C,
Jarreau PH, Régnier B.
The organisation of intensive care in a situation of pandemic avian
influenza.
Rev Mal Respir, 2008, 25:223.
Bialecki E, Paget C, Fontaine J, Capron M, Trottein F, and Faveeuw C.
Role of marginal zone B lymphocytes in invariant Natural Killer T cell
activation.
J Immunology, 2009, 82: 6105-6113.
Henry E, Desmet CJ, Garzé V, Fiévez L, Bedoret D, Heirman C, Faisca
P, Jaspar FJ, Gosset P, Jacquet AP, Desmecht D, Thielemans K, Lekeux
P, Moser M, Bureau F.
Dendritic cells genetically engineered to express IL-10 induce long-lasting
antigen-specific tolerance in experimental asthma.
J Immunol, 2008, 181:7230-7242.
Boutoille D, Marechal X, Pichenot M, Chemani C, Guery B, Faure K.
FITC-albumin as a marker for assessment of endothelial permeability in
mice: comparison with 125I-albumin.
Exp Lung Res, 2009, 35:263-271.
Kipnis E, Hansen K, Sawa T, Moriyama K, Zurawel A, Ishizaka A,
Wiener-Kronish J.
Proteomic analysis of undiluted lung epithelial lining fluid.
Chest, 2008,134:338-345.
Carnoy C, Roten CA.
The dif/Xer recombination systems in proteobacteria.
PLoS One, 2009, 4:e6531.
Chemani C, Imberty A, de Bentzmann S, Pierre M, Wimmerová M,
Guery BP, Faure K.
Role of LecA and LecB lectins in Pseudomonas aeruginosa-induced lung
injury and effect of carbohydrate ligands.
Infect Immun, 2009, 77:2065-2075.
Le Berre R, Nguyen S, Nowak E, Kipnis E, Pierre M, Ader F, Courcol R,
Guery BP, Faure K. Pyopneumagen Group.
Quorum-sensing activity and related virulence factor expression in
clinically pathogenic isolates of Pseudomonas aeruginosa.
Clin Microbio Infect, 2008, 14:337.
Secher T, Lacas-Gervais S, Gratadoux JJ, Lafont F, Dagorn JC,
Ryffel B, Akira S, Langella P, Nùñez G, Sirard JC, Iovanna J, Simonet
M, Chamaillard M.
Toll-like receptor 2 is critical for induction of Reg3 beta expression and
intestinal clearance of Yersinia pseudotuberculosis.
Gut, 2009, 58:771-776.
Mokart D, Kipnis E, Guerre-Berthelot P, Vey N, Brun JP, Blache JL,
Mege JL, Blaise D, Guery BP.
Monocyte deactivation in neutropenic acute respiratory distress
syndrome patients treated with granulocyte colony-stimulating factor.
Crit Care, 2008, 12:R17.
Nempont C, Cayet D, Rumbo M, Bompard C, Villeret V, Sirard JC.
Deletion of Flagellin’s Hypervariable Region Abrogates Antibodymediated neutralization and systemic activation of TLR5-dependent
immunity.
J Immunol, 2008, 181:2036.
Estrella C, Rocks N, Paulissen G, Quesada-Calvo F, Noël A, Vilain E,
Lassalle P, Tillie-Leblond I, Cataldo D, Gosset P.
Role of A disintegrin and metalloprotease-12 in neutrophil recruitment
induced by airway epithelium.
Am J Respir Cell Mol Biol, 2009, 41:449-458.
Rocks N, Estrella C, Paulissen G, Quesada-Calvo F, Gilles C, Guéders
MM, Crahay C, Foidart JM, Gosset P, Noel A, Cataldo DD.
The metalloproteinase ADAM-12 regulates bronchial epithelial cell
proliferation and apoptosis.
Cell Prolif, 2008, 41:988-1001.
Galperine T, Flateau C, Venon MD, Lescure FX, Béraud G,
Said Ibrahim T, Delisle F, Durand F, Faure K, Pialoux G, Guery B.
Recurrent bacteremia, a complication of cyanoacrylate injection for
variceal bleeding: report of two cases and review of the literature.
Case Report Med, 2009, 407053.
Roumier T, Capron M, Dombrowicz D, and Faveeuw C.
Pathogen induced regulatory cell populations preventing allergy through
the Th1/Th2 paradigm point of view.
Immunol Res, 2008, 40:1.
Ganter MT, Roux J, Su G, Lynch SV, Deutschman CS, Weiss YG,
Christiaans SC, Myazawa B, Kipnis E, Wiener-Kronish JP, Howard M,
Pittet JF.
Role of small GTPases and alphavbeta5 integrin in Pseudomonas
aeruginosa-induced increase in lung endothelial permeability.
Am J Respir Cell Mol Biol, 2009, 40(1):108-118.
Tillie-Leblond I, J de Blic, F Jaubert, B Wallaert, P Scheinmann,
P Gosset.
Airway Remodeling is correlated with obstruction in children with severe
asthma.
Allergy, 2008, 63:533.
Janot L, Sirard JC, Secher T, Noulin N, Fick L, Akira S, Uematsu S,
Didierlaurent A, Hussell T, Ryffel B, Erard F.
Radioresistant cells expressing TLR5 control the respiratory epithelium's
innate immune responses to flagellin.
Eur J Immunol, 2009, 39:1587-1596.
49
Contents
Moreau C, Gosset P, Brunaud-Danel V, Lassalle P, Degonne B, Destee
A, Defebvre L, Devos D.
CSF profiles of angiogenic and inflammatory factors depend on the
respiratory status of ALS patients.
Amyotroph Lateral Scler, 2009, 10:175-181.
Torres D, Dieudonné A, Ryffel B, Vilain E, Si-Tahar M, Pichavant M,
Lassalle P, Trottein F, Gosset P.
Double-Stranded RNA Exacerbates Pulmonary Allergic Reaction through
TLR3 : Implication of Airway Epithelium and Dendritic Cells.
J Immunol, 2010, 185:451-459.
Paget, C., Bialecki, E., Fontaine, J., vendeville, C., Mallevaey, T.,
Faveeuw, C and Trottein, F.
Role of invariant Natural Killer T cells in immune responses to CpG
oligonucleotides.
J Immunol, 2009, 182:1846-1853.
Janot L, Erard F, Bertout J, Leger H, Sebbane F, Benecke A,
Renauld JC, Hardt WD, Ryffel B, Sirard JC.
TLR5 Signaling Stimulates the Innate Production of IL-17 and IL-22 by
CD3negCD127+ Immune Cells in Spleen and Mucosa.
J Immunol, 2010, 185:1177-1185.
Sirard JC, Didierlaurent A, Cayet D, Sierro F, Rumbo M.
Toll-like receptor 5- and lymphotoxin beta receptor-dependent epithelial
Ccl20 expression involves the same NF-kappaB binding site but distinct
NF-kappaB pathways and dynamics.
Biochim Biophys Acta, 2009, 1789:386-394.
PhD
François VANHOUTTE
Directeur de thèse : F. Trottein
« Rôle des récepteurs Toll-like dans les interactions hôte/schistosome »
Université de Lille 2. Discipline Immunologie
15 Juin 2007
Taront S, Dieudonné A, Blanchard S, Jeannin P, Lassalle P,
Delneste Y, Gosset P.
Implication of scavenger receptors in the interactions between diesel
exhaust particles and immature or mature dendritic cells.
Part Fibre Toxicol, 2009, 13:6-9.
Laetitia BREUILH
Directeur de thèse : I. Tillie-Leblond
« Rôle des récepteurs Toll-like et des récepteurs lectiniques dans la réponse
immune de type 2 : modèle de la schistosomiase »
8 Octobre 2007
Trottein F, Schaffer L, Ivanov S, Paget C, Vendeville C, Cazet A,
Groux-Degroote S, Lee S, Krzewinski-Recchi MA, Faveeuw C, Head
SR, Gosset P, Delannoy P.
Differential glycosyltransferase and sulfotransferase gene expression
profiles in human monocytes, dendritic cells and macrophages.
Glycoconjugate J, 2009, 26:1259-1274.
Rozen LEBERRE
Directeur de thèse : B. Guery
Physiopathologie et rôle des facteurs de virulence dans la pneumonie à
Pseudomonas aeruginosa.
Université de Marseille
15 Octobre 2007
2010
Adamczyk S, Robin E, Simerabet M, Kipnis E, Tavernier B, Vallet B,
Bordet R, Lebuffe G.
Sevoflurane pre- and post-conditioning protect the brain via the
mitochondrial K ATP channel.
Br J Anaesth, 2010, 104:191-200.
Eric KIPNIS
Directeur de thèse : B. Guery
Place du SST III dans l’atteinte infectieuse.
Université de Lille 2, Sciences de la vie et de la santé
13 Décembre 2007
Beauvillain C, Meloni F, Sirard JC, Blanchard S, Jarry U, Scotet M,
Magistrelli G, Delneste Y, Barnaba V, Jeannin P.
The scavenger receptors SRA-1 and SREC-I cooperate with TLR2 in
the recognition of the hepatitis C virus non-structural protein 3 by
dendritic cells.
J Hepatol, 2010, 52:644-51.
Solenne TARONT
Directeur de thèse : P. Gosset
« Impact des motifs bactériens et des particules de diesel sur les
interactions épithélium bronchique / cellules dendritiques »
3 Octobre 2008
DeKruyff RH, Bu X, Ballesteros A, Santiago C, Chim YL, Lee HH,
Karisola P, Pichavant M, Kaplan GG, Umetsu DT, Freeman GJ,
Casasnovas JM.
T cell/transmembrane, Ig, and mucin-3 allelic variants differentially
recognize phosphatidylserine and mediate phagocytosis of apoptotic
cells.
J Immunol. 2010, 184:1918-30.
Patent
Marichal T, Bedoret D, Mesnil C, Pichavant M, Goriely S, Trottein F,
Cataldo D, Goldman M, Lekeux P, Bureau F, Desmet CJ.
Interferon response factor 3 is essential for house dust mite-induced
airway allergy.
J Allergy Clin Immunol, 2010, Jul 29.
Sirard JC
Novel immunoadjuvant flagellin-based compounds and use thereof
25 juin 2008 – EP n°08 305 327.2, 23 juin 2009 - PCT/EP2009/057836,
30 décembre 2009 - WO2009/156405
Muñoz N, Van Maele L, Marqués JM, Rial A, Sirard JC, Chabalgoity JA.
Mucosal administration of flagellin protects mice from Streptococcus
pneumoniae lung infection.
Infect Immun, 2010,
Renneson J, Guabiraba R, Pereira-Silkva RE, Ivanov S, Fontaine J,
Paget C, Ryffel B, Quesniaux V, Faveeuw C, Teixeira M, Trottein F.
A detrimental role for invariant Natural Killer T cells in the pathogenesis
of dengue virus infection.
J Immunol, 2010, in press.
50
Contents
cytokine production after direct in vitro stimulation of human
peripheral blood mononuclear cells (PBMC), although variable
and sometimes even opposite immuno-modulatory properties
were noted. We observed moreover, that the in vitro cytokine
expression profiles correlated nicely with the capacity to
protect mice from an experimental 2,4,6-trinitrobenzenesulfonic acid (TNBS)-induced colitis and established that
this protection was strain-specific and dose-dependant.
In a further mechanistic follow-up study, we have demonstrated
that this selective protection against intestinal inflammation
could be achieved through the systemic route, i.e. after intraperitoneal administration of LAB, suggesting a mechanism that
may act at distance and involve the migration of regulatory cell
populations. As a result we established that certain lactobacilli
are able to differentially activate bone marrow derived dendritic
cells (BMDCs), leading to the generation of regulatory DCs,
which confer protection against colitis upon adoptive transfer
to TNBS-treated mice. Our results further suggested that the
anti-inflammatory effect depends on the activation of
regulatory T cells and the Indolamine 2, 3 dioxygenase immunesuppressive pathway. By the use of BMDCs derived from
deficient mice (KO), we could further show that this effect was
TLR2 and Nod2-dependent. It is known that Nod1 and Nod2
(also known as CARD15 and CARD4, respectively) sense
intracellularly microbial-associated molecular patterns (MAMPs)
and activate the NF-κB immunoregulatory pathway, important
in inflammation. Conversely, NOD2 has also been shown to be
a negative regulator of the TLR2-dependent NF-κB signalling
cascade. It is striking that clinical trials with probiotic bacteria
on patients with Crohn’s disease have rarely been effective,
especially now that it is known that mutations in the Nod2 gene
are a confirmed risk factor for Crohn’s disease. Using the same
TNBS colitis model in Nod2 KO mice, we participated in a study
(collaboration with JP Hugot, Hopital H. Debré, Paris), which
showed that card15/nod2 deficiency induced an abnormal
development and function of Peyer’s patches, characterized by
an exaggerated immune response and an increased intestinal
permeability. In the framework of the CIIL there is a clear interest
in studying the role of these genes in the bacteria-host signalling.
Earlier results indicated a possible pivotal role of certain cellwall components in the selective immunomodulation
potential of LAB, as some cell-wall mutants of a Lactobacillus
plantarum reference strain were shown to increase
significantly their anti-inflammatory properties in the in vitro
and in vivo models used (collaboration with P. Hols and
J. Delcour, Université Catholique de Louvain, Belgique).
Further research, using purified fractions of peptidoglycan
and proteins is now underway and initial evidence was
obtained that structurally different peptidoglycans are indeed
responsible for some of the observed differences in the antiinflammatory potential of probiotics.
Using the available models and the selected strains,
characterized in full, we will further extend this research topic
and continue to study the interaction between the bacteria
and the intestinal mucosa by identifying the signals and the
cellular populations involved in this cellular crosstalk. A special
interest will be paid to the characterization of different
compounds of the bacterial cell-wall (collaboration with the
team of Ivo Boneca, Institut Pasteur, Paris, France and Thomas
Hartung, University of Konstanz, Germany). In collaboration
with the team of Martine Heyman (Hopital Necker, Paris,
France) we studied e.g. the anti-inflammatory capacity of
metabolites produced by a Bifidobacterium strain. We also
participated in a collaborative study (P. Langella, INRA, Jouy-
Lactic Acid Bacteria
and Mucosal Immunity
Bruno POT, DR IPL
Group Members :
Daniel-Desseyn Catherine, CR1 IPL
Foligné Benoit, CR1 IPL
Grangette Corinne, CR2 IPL
Breton Jérôme, CE1 IPL
Dennin-Peucelle Véronique / Véronique Valenti IE1 IPL
Dewulf Joëlle, IE1 IPL
Abboud George, Postdoctoral fellow IPL
Goudercourt-Boutillier Denise, Technician IPL
Poiret Sabine, Technician IPL
Macho Fernandez Elise, PhD Student MENRT
Key words :
Lactic acid bacteria. Probiotics. Mucosal immunity. Immunemodulation. Live vehicle. Food safety. Functional foods.
Gastro-intestinal diseases. Gut homeostasis. Inflammation
models. Infection models. Heavy metal detoxification.
A - Research Report
The LABMI team is currently a team supported by the Institut
Pasteur de Lille and has a major interest in health applications
of lactic acid bacteria (LAB). In an attempt to substantiate the
effectiveness of the so-called ‘probiotic’ LAB, we have developed
and used different standardized in vitro and in vivo models.
While screening immune-modulation properties we observed
strong strain-specific differences, yielding strains with highly
different properties that was used as the basic material for a
number of comparative research projects aiming at
understanding the underlying mechanisms of action. Knowledge about mechanisms is of interest to the industry and the
medical community, as it facilitates the definition of suitable
biomarkers and will guide the selection process for strains for
improved, dedicated health applications. A second research axis
of the laboratory focuses on the demonstration that recombinant LAB can efficiently deliver vaccine antigens and therapeutic
molecules. The direct delivery of vaccines or therapeutic
molecules at the host’s mucosal surfaces by safe LAB with
inherent adjuvant properties has led to promising perspectives
for their development as prophylactic or therapeutic tools. The
lab has also shown that the activities of selected LAB can be
enhanced by genetic modification, for example by changing
selected target structures at the bacterial surface.
1. Immuno-modulatory properties of
lactic acid bacteria
In the context of a previous European network (Deprohealth,
N° QLK-CT2000-00146) and in collaboration with industrial
partners, we have studied a large collection of LAB, which are
thought to have beneficial properties for the host. In order to
understand the differences between pathogenic bacteria on
the one hand and commensal or therapeutic bacteria on the
other hand, we studied in vitro the immune-modulation
properties of a large collection of these LAB. We could show
that various LAB strains are able to differentially modulate
51
Contents
en-Josas) aiming to characterize of the anti-inflammatory
capacities of Faecalibacterium prausnitzii, a member of the
Firmicutes from the commensal flora and for which a reduced
level has been associated with increased risk of postoperative
recurrence of ileal Crohn disease.
The scientific and economic viability of the approach will
hopefully be confirmed during an ongoing clinical trial at the
Catholic University Leuven, Belgium, under the supervision of
Dr. P. Rutgeers and Dr. K. Verbeke. In this trial, we are currently
investigating the efficacy of a probiotic strain selected by our
team, in patients suffering from pouchitis and ulcerative colitis.
In collaboration with J. Pestel (Inserm U 416, Institut Pasteur de
Lille), we also showed that selected probiotic strains can inhibit
the in vitro production of Th2 cytokines IL-4 and IL-5 after
stimulation of PBMC’s of allergic patients. In the framework of
two other EU research projects (LABDEL and Interreg-III) we
demonstrated that mucosal co-application of LAB and wellknown allergens (the birch pollen allergen Bet v 1 in
collaboration with the team of U. Wiedermann, University
of Vienna, Austria and the major house dust mite allergen Der
p 1 of Dermatophagoides pteronyssinus, in collaboration
with A. Jacquet, University of Brussels, Belgium) induced a
T helper type 1 (Th1) biased allergen specific response.
possibility of improving the observed adjuvanticity of LAB by
modifying the LAB cell wall has led to a patent and was also
illustrated in a vaccine model against Helicobacter pylori in
collaboration with the team of B. Corthesy (Lausanne, Suisse).
The successful application of recombinant LAB strains in
different animal models has led to their further evaluation as live
vehicles to deliver human therapeutic molecules such as
allergens. As mentioned above, we first demonstrated that
recombinant LAB strains producing Bet v 1 and Der p 1 were able
to shift in vivo the specific Th2 allergic immune response towards
a Th1 anti-allergic immune response. These observations were
only possible when the allergen and the LAB were administered
via the same route of administration. In order to assure an
efficient delivery we constructed different LAB strains, which
secreted or produced the intact protein Bet v 1 and the protein
pro-Der p 1. Using these constructs, interesting results were
obtained in mouse models of allergy, confirming the potential
of LAB to deliver therapeutic molecules at the mucosal level.
In a very successful collaboration with the team of M. Simonet,
Inserm U801, we recently developed new expression vectors
to secrete anti-inflammatory molecules in the digestive tract
such as the immunomodulatory LcrV (Low calcium responsive
V antigen) protein naturally secreted by pathogenic Yersinia.
We demonstrated that recombinant LAB strains producing
LcrV protected and treated mice against experimentally
induced intestinal inflammation in a TLR-2 and IL-10
dependent manner. This novel approach will be further
elaborated in future projects which will highlight the potential
of (pathogen-derived) immune-molecules delivered by LAB,
used as novel therapeutics in inflammatory bowel disease.
Similarly we evaluated the potential of the recombinant
Lactococcus lactis strain secreting the Yersinia pseudotuberculosis low-calcium response V (LcrV) antigen for mucosal
vaccination against Yersinia infections. We showed that the
recombinant strain induced specific systemic and mucosal
antibody and cellular immune responses after intranasal
immunization and protected mice against both oral and
systemic Y. pseudotuberculosis infections. This constituted the
first proof of principle for a novel anti-Yersinia mucosal
vaccination strategy using recombinant lactic acid bacteria.
2. Safety properties of lactic acid
bacteria
Despite the fact that LAB are generally known to be safe and
used extensively in many food and feed applications, a limited
number of LAB strains have been associated with endocarditis,
sepsis and abscesses. During a European research project on
probiotic safety (ProSafe) a collection of ‘clinical’ isolates of LAB
was characterized. From the project coordinator (Prof. H.
Goossens, University Antwerp, Belgium) we obtained some
well-characterized strains isolated from clinical material. Using
these strains (already resistant to multiple antibiotics) and four
well-selected probiotic reference strains from our probiotic
collection (artificially labelled with multiple antibiotic
resistances), we measured the bacterial translocation from the
gut to extra-intestinal organs in the above-mentioned TNBSinduced colitis model. This study confirmed that LAB in
general do NOT translocate from the gut in normal healthy
mice, but revealed a consistent translocation and worsening
of colitis for two ‘pathogenic’ strains of Lactobacillus in mice
that had been treated with very high doses of TNBS. The
probiotic control strains, however, did not translocate under
these conditions and reduced the inflammation caused by the
TNBS instillation. These experiments confirmed that the safety
and functionality of LAB is strain dependent. These results
clearly demonstrate that although European QPS guidelines
can be followed, some additional safety criteria may be
investigated and results added to the probiotic-supporting
dossier of the strain studied.
4. Lactic acid bacteria to fight pathogens and obesity
There are many indications that LAB can have negative growth
effects on pathogenic bacteria. During a former Marie-Curie
fellowship we have developed an in vivo infection model for
Salmonella (in collaboration with the team of M. Simonet, Inserm
U801). Using a positive reference strain of Lactobacillus
fermentum, selected for its in vitro activity against Salmonella, we
set up a mice model to confirm in vivo anti-Salmonella activity.
Using this Salmonella typhimurium infection model it was
possible to study the immunological as well as the metabolic
and anti-microbial aspects of potential probiotic strains. In a later
stage, we set up a second murine infection model with the
pathogen Citrobacter rodentium, where we can measure the antiinflammation and anti-infection potential of a given probiotic or
mixture of strains. This model, which induces colitis and causes
infection, allows us to study various mechanistic aspects of an
intervention with selected strains and to further explore, for
example, the role of oxidative stress in infection and
inflammation and to perhaps demonstrate a possible protective
role of LAB through their anti-oxidant capacities.
3. Use of lactic acid bacteria as vaccine
and therapeutic delivery vehicles
Within two successive European networks (LABVAC) we
explored the potential of LAB to efficiently deliver antigens, (i.e.
Tetanus toxin fragment C) to mucosal surfaces and to induce
protective immune responses. We notably demonstrated that
the use of recombinant mutant strains was more efficient in
delivering antigens and in enhancing immune responses. The
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Contents
allow us to gain better insight in the physical aspects of host
- bacteria interaction and may be an interesting tool to follow
the faith of different LAB strains after ingestion.
development 2010
Publications
The intestine is a complex and dynamic ecosystem containing
about one thousand bacterial species, comprising potential
pathogens, commensal bacteria or microorganisms beneficial
for the health of the host. This microbiota establishes shortly
after birth and is essential in priming the immune system, in
contributing to gut homeostasis and in intestinal functionality.
The emergence of a number of immunological disorders, such
as Th2-driven allergic diseases or disorders driven by Th1
effector cells, like inflammatory bowel diseases (IBD), type 1
diabetes and multiple sclerosis, have been attributed to a
modern lifestyle linked to certain dietary, hygienic and medical
habits. Notably, IBD was suggested to be associated with
abnormal immune responses and a breakdown of tolerance
against the indigenous microflora. It has been shown recently
that recognition of commensal bacteria by Toll like receptors
(TLR) at the host cell surface plays a crucial role in the
maintenance of intestinal homeostasis. Largely used in a variety
of food fermentation processes, certain lactic acid bacteria (LAB)
are well known to have health benefits in humans or animals.
The proposal of probiotics in a medical context has recently
gained a lot of momentum, and therapeutic / prophylactic
capacities have been demonstrated both in animal models and
in clinical studies. Despite some successes, a number of studies
did not yield significant results. The failure of these studies in
many cases can be linked to the lack of a proper strain selection
prior to the trial and to the definition of endpoints, which are
defined according to criteria that apply to clinical trials with
pharmaceutical endpoints. The use of probiotics as an add-on
therapy in immune disorder treatments will in many cases
prove to be very rewarding, given that adapted strains and
proper administration / delivery protocols are used.
In this respect we are currently expanding our research field
to study in depth the mechanisms underlying the observed
link between peptidoglycan type and anti-inflammation. This
could lead to the development of potent anti-inflammatory
preparations that can be used in a prophylactic approach to
fight IBD. In the meantime we hope to obtain positive results
from the ongoing clinical trial in Belgium, where a
Bifidobacterium breve strain is used to extend remission time
in patients with ulcerative colitis and pouchitis.
More recently we started to explore to which extend the antiinflammatory activity of probiotic strains might be useful in
the therapy of obesity (collaboration with Isabelle Wolowczuk,
CNRS) and in reducing the development of glucose tolerance.
In an attempt to better understand the observed efficiency of
selected probiotics to treat active phases of Clostridium difficile
diarrhea, we are currently optimizing a third infection model
that will allow us to compare different strains and to study the
underlying mechanisms of the protection.
We furthermore extend the possible application fields of LAB
in the framework of a national ANR research project (ANR 09CESA-016) that will study the potential of probiotics, bacteria
and yeast, to reduce the toxicity of heavy metals like cadmium
and lead, taken up from the environment or through the diet.
Finally, in 2010 we will also optimize the luciferase labeling of
LAB, allowing to detect and follow live microorganisms in
/along the digestive tract of small animals. This project will
2007
Barreau F, Meinzer U, Chareyre F, Berrebi D, Niwa-Kawakita M,
Dussaillant M, é B, Ollendorff V, Heyman M, Bonacorsi S, Lesuffleur T,
Sterkers G, Giovannini M, Hugot JP.
CARD15/NOD2 is required for Peyer's patches homeostasis in mice.
Plos One, 2007, 2:e523.
Daniel C, Repa A, Mercenier A, Wiedermann U, Wells J.
The European LABDEL project and its relevance to the prevention and
treatment of allergies.
Allergy, 2007, 62:1237-1242.
De Bruyne V, Al-Mulla F, Pot B.
Methods for microarray data analysis.
Meth Mol Biol, 2007, 382:373-391.
Foligné B, Dessein R, Marceau M, Poiret S, Dewulf J, Pot B, Simonet,
Daniel C.
Therapeutic potential of Yersinia anti-inflammatory components.
Adv Exp Med Biol, 2007, 603:361-366.
Foligné B, Dessein R, Marceau M, Poiret S, Chamaillard M, Pot B,
Gastroenterol, 2007, 133:862-874.
Foligné B, Nutten S, Grangette C, Dennin V, Goudercourt D,
Poiret S, Dewulf J, Brassard D, Mercenier A, Pot B.
Correlation between in vitro and in vivo immunomodulatory properties
of lactic acid bacteria.
Foligné B, Zoumpopoulou G, Dewulf J, Ben Younes A, Chareyre F,
Sirard JC, Pot B, Grangette C.
A key role of dendritic cell in probiotic functionality.
Plos One, 2007, 2:e313.
Grangette C.
Probiotiques et régulation de la réponse immune allergique et
inflammatoire.
Cah Nutr Diét, 2007, 42: 2S76-2S85.
Hisbergues M, Magi M, Rigaux P, Steuve J, Garcia L, Goudercourt D,
Pot B, Pestel J, Jacquet A.
In vivo and in vitro immunomodulation of Der p 1 allergen-specific
response by Lactobacillus plantarum bacteria.
Clin Exper Allergy, 2007, 37:1286-1295.
Perea Vélez M, Verhoeven T, Draing C, Von Aulock S, Pfitzenmaier
M, Geyer A, Lambrichts I, Grangette C, Pot B, Vanderleyden J, De
Keersmaecker SCJ.
Functional Analysis of D-alanylation of Lipoteichoic Acid in the Probiotic
Strain Lactobacillus rhamnosus GG.
Appl Environ Microbiol, 2007, 73:3595-3604.
Pot B, Gillis M.
The genus Aquaspirillum.
In The Prokaryotes Vol. 5: Proteobacteria: Alpha and Beta Subclasses.
Dworkin, M. Falkow, S.; Rosenberg, E.; Schleifer, K.-H.; Stackebrandt, E.
(Eds.) 3rd ed. Springer. 2007, p 710-722.
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Contents
Ratajczak C, Duez C, Grangette C, Pochard P, Tonnel AB, Pestel J.
Impact of lactic acid bacteria on dendritic cells from allergic patients in
an experimental model of intestinal epithelium.
J Biomed Biotechnol, 2007, Article ID 71921, 9 pages doi:10.
1155/2007/71921.
Heuvelin, E, Lebreton C, Grangette C, Pot B, Cerf-Bensussan N,
Heyman M.
Mechanisms Involved in Alleviation of Intestinal Inflammation by
Bifidobacterium breve Soluble Factors.
PloS ONE, 2009, 4:e5184.
Rigaux P, Daniel C, Pot B, Pestel J, Jacquet A.
Immunomodulation of house dust mite allergy by a recombinant lactic
acid bacteria producing Der p 1.
Allergy, 2007, 62: 245.
Pirnay, JP; Bilocq, F; Pot, B, Cornelis, P, Zizi, M, Van Eldere J,
Deschaght P, Vaneechoutte M, Jennes S, Pitt T, De Vos
Pseudomonas aeruginosa Population Structure Revisited.
PloS ONE, 2009, 4:e7740.
2008
Pot B, Tsakalidou E
Taxonomy and metabolism of Lactobacillus. Chapter 2 In: Lactobacillus
Molecular Biology.
Edited by : Ljungh and Wadstrom, 2009, Caister Academic press, p 3- 58.
Mattarelli P, Bonaparte C, Pot B, Biavati B.
Proposal to reclassify the three biotypes of Bifidobacterium longum as
three subspecies: Bifidobacterium longum subsp. longum subsp. nov.,
Bifidobacterium longum subsp. infantis comb. nov. and Bifidobacterium
longum subsp. suis comb. nov.
Int J Syst Evol Microbiol, 2008, 58:767-777.
Rigaux P, Daniel C, Hisbergues M, Muraille E, Hols P, Pot B, Pestel J,
Jacquet A.
Immunomodulatory properties of Lactobacillus plantarum and its use as
a recombinant vaccine against mite allergy.
Allergy, 2009, 64:406-414.
Pot B.
The taxonomy of lactic acid bacteria. In. Bactéries lactiques, de la
génétique aux ferments.
G. Corrieu, F.M. Luquet, eds. Lavoisier, 2008, p. 1-152.
Zoumpopoulou G, Tsakalidou E, Dewulf J, Pot B, Grangette C.
Differential crosstalk between epithelial cells, dendritic cells and bacteria
in a co-culture model.
Int J Food Microbiol, 2009, 131:40-51.
Roussel Y, Daniel C.
Bactéries lactiques utilisées comme vecteurs de délivrance de molécules
vaccinales et thérapeutiques aux voies muqueuses. Les bactéries lactiques :
Physiologie, Métabolisme, Génomique et Applications Industrielles des
bactéries lactiques.
Editions Economica. 2008, In press (non traduit).
2010
Asteri IA, Papadimitrio K, Boutou E, Anastasiou R, Pot B, Vorgias CE,
Tsakalidou E.
Characterization of pLAC1, a cryptic plasmid isolated from Lactobacillus
acidipiscis and comparative analysis with its related plasmids.
Sokol H, Pigneur B, Watterlot L, Lakhdari O, Gratadoux JJ, Blugeon S,
Bridonneau C, Furet JP, Corthier G, Grangette C, Vasquez N, Pochar Pt,
Trugnan G, Thomas G, Blottiere HM, Doré J, Marteau P, Seksik P,
Langella P.
The commensal Firmicute Faecalibacterium prausnitzii exhibits in vitro
and in vivo anti-inflammatory effects with potential applications for
Crohn's disease.
Proc Natl Acad Sci U S A, 2008, in press.
Cousin F.J., Foligne B.,Jan G.
Dairy propionibacteria as human probiotics : recent evidences.
Review, Dairy Sci Technol, 2010, In press.
De Vuyst L, Vincent P, Makras E, Leroy F, Pot B.
Peptide Extracts from Cultures of Certain Lactobacilli Inhibit Helicobacter
pylori.
Probiotics & Antimicro Prot, 2010, 2:26–36.
Vankerckhoven, Huys G, Vancanneyt M, Vael C, Klare I ,Romond MB,
Entenza JM, Moreillon P, Wind RD, JKnol J, Wiertz E, Pot B,
Vaughan EE, Kahlmeter G, Goossens H.
Biosafety assessment of probiotics used for human consumption:
recommendations from the EU-PROSAFE project.
Trends in Food Sci Technol, 2008, 19:102-114.
Foligne B, Dewulf J, Vandekerckove P, Pignede G, Pot B.
Probiotic yeasts: anti-inflammatory potential of various non-pathogenic
strains in experimental colitis in mice.
Wolowczuk I, Verwaerde C, Viltart CO, Delanoye A, Delacre M, Pot B,
Grangette C.
Feeding our immune system: impact on metabolism.
Clin Develop Immunol, 2008, 639:803.
Foligne B, Dewulf J, Vandekerckove P, Pignede G, Pot B.
Probiotic yeasts : anti-inflammatory potential of various non-pathogenic
strains in experimental colitis in mice.
World J Gastroenterol, 2010,16:2134-2145.
Zoumpopoulou G, Foligne B, Christodoulou K, Grangette C, Pot B,
Tsakalidou E.
Lactobacillus fermentum ACA-DC 179 displays probiotic potential in vitro
and protects against TNBS-induced colitis and Salmonella infection in
murine models.
Foligné B, Dewulf J, Breton J, Claisse O, Lonvaud-Funel A, Pot B.
Probiotic properties of non-conventional lactic acid bacteria :
Immunomodulation by Oenococcus oeni.
Mattocks C.J, Watkins G, Ward D, Janssens T, Bosgoed E AJ, van der Donk
K, Ligtenberg M JL, Pot B, Theelen J, Cross N CP, Scheffer H, Matthijs G
Inter-laboratory Diagnostic Validation of Conformation Sensitive
Capillary Electrophoresis for Mutation Scanning.
Clin Chem, 2010, 56:593-602.
2009
Daniel C, Sebbane F, Poiret S, Goudercourt D, Dewulf J, Mullet C,
Simonet M, Pot B.
Protection against Yersinia pseudotuberculosis infection conferred by a
Lactococcus lactis mucosal delivery vector secreting LcrV.
Vaccine, 2009, 27:1141-1144.
O’Flaherty S, Saulnier DM, Pot B, Versalovic J.
How Can Probiotics and Prebiotics Impact Mucosal Immunity ?
Gut Microbes, 2010, 1:5, 1-8.
Foligne, B, Pot, B.
Comment on "Some immunomodulatory effects of probiotic bacteria
might be due to porcine neutrophil elastase inhibitor, a serpin present in
MRS broth".
Immunology Letters, 2009, 122:227-228.
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Contents
the interaction of overlapping complex responses and
signals of the immune system, including T-cells and
cytokine/chemokine networks, which may play a central role
in the outcome of the disease. The diversity of the responses
induced may depend on the complex composition of the
parasite (including antigenic, superantigenic and mitogenic
activities) that may lead to an overstimulation of the immune
system. Using experimental models, C57BL/6 (B6) mice
infected with either Plasmodium (P.) yoelii (protective
responses) or P. berghei ANKA (pathological responses) and
cohorts of P. falciparum infected patients, we analyzed
lymphocyte populations involved in the different immune
responses to determine their phenotype as well as their
repertoire diversity, their dynamic and their function at
physiological state and during pathogenesis.
Basic and Clinical
Immunology of Parasitic
Diseases
Sylviane PIED, DR2 CNRS
Group Members :
Hermann Emmanuel, MCU Univ Lille 2
Riveau Gilles, DR2 CNRS
Roland Jacques, CR1 CNRS
Cazenave Pierre-André, Pr Emérite Univ Paris 6
Herbert Fabien, IE2 CNRS (CDD)
Schacht Anne-Marie, IE IPL
Bansal Devendra, Postdoctoral fellow CEFIPRA
Shrivastava Sandeep, Postdoctoral fellow CNRS
Gaayeb-Juillian Lobna, PhD Student Univ Lille 2
Hannon Jean-baptiste, Master 2 Student Univ Lille 2
1.1 Immune responses involved in protection against
primary P.yoelii infection.
Most studies exploring protective immune responses during
Plasmodium infection focused on reactions to immunization
with irradiated sporozoites or candidate vaccines. Little is
known about the wide variety of responses induced by
primary infection. In addition, our understanding of protective
immunity remains incomplete. We used the experimental
model of B6 mice infected with P. yoelii 265 By. The mice
develop parasitemia that is rapidly and spontaneously cured.
The advantage of using the P. yoelii model is that infection can
be initiated by injecting either sporozoites, to obtain a
complete cycle of plasmodium within the mammalian host
(intrahepatic schizonts and blood stages) or by injecting blood
stages. Thus, this system enables differentiate between
responses specific to liver stages or blood stages. In this
model, we analysed the immune response both at cellular and
at molecular levels. We determined the kinetics and
phenotypes of responding T cells and analyzed their functions
during infection. Our studies mainly focus on the intrahepatic
T cells. We demonstrated these last years that nonconventional natural killer αβ (NKT) cells, a major T
lymphocyte population in the liver that recognizes glycolipids
presented by a non–classical MHC class I molecule, CD1d,
highly conserved among species, are implicated in the first
defense mechanisms against P. yoelii liver stage. The NKT cells
elicited in P. yoelii-infected B6 mouse livers are heterogeneous
and composed of CD1d-dependent invariant CD4+ NKT cells
and of CD4CD8 double-negative (DN) NKT cells, which include
CD1d-dependent invariant and non-invariant cells, and CD1dindependent cells (Soulard et al., 2007). During infection,
parasite-activated hepatic DN NKT cells secrete interferon
(IFN)γ and tumor necrosis factor (TNF)α (Th1 cytokines).
These cells isolated from infected mice inhibited intra-hepatic
development of the parasite in vitro in a CD1d-dependent
manner. Pertinently, the parasite burden was higher in the
livers of B6.CD1d-deficient (no NKT cells) than in those of B6
mice demonstrating thus that NKT cells and CD1d may be
involved in the early control of primary P. yoelii infection.
Clearly we demonstrate that innate immune responses
mediated by NK and NKT cells participate in early immune
mechanisms that control Plasmodium infection mainly by
acting on Plasmodium pre-erythrocytic stages (Roland, 2006;
Soulard, 2007). One of the common factors of the P. yoelii
induced NK and NKT cells response is their release of IFNγ
upon in vivo stimulation during infection. Consequently, we
examined the kinetics of pro-inflammatory cytokines such
Key words :
Malaria, Schistosomiasis, immune responses, immunopathology, immunoregulation, genetic control, clinical immunology,
immuno-epidemiology, vaccine strategies, clinical trials, Africa,
Asia.
A - Research Report
The BCIP team results from the fusion of the “Malaria
Immunopathophysiology” group 1 headed by Sylviane Pied
from the Infectious Immunophysiopathology Unit, CNRS URA
1961, Department of Immunology at the Institut Pasteur in
Paris and the "Clinical Parasite Immunology" group 2 headed
by Gilles Riveau working on schistosomiasis and malaria in
Africa and Emmanuel Herman, both from Inserm U 547,
Institut Pasteur de Lille. The team 1 has joined the Institut
Pasteur of Lille in January 2008 and was supported by the
“Programme d’Incitation à la Mobilité d’Equipe” PIME) of CNRS
and Inserm U 547.
1. Malaria Immunophysiopathology
The malaria parasite life cycle involves 2 obligatory hosts,
and infection results from intricate interactions between the
parasite and the mosquito, and then man. Because these 2
hosts are separated by hundreds of millions of years of
divergent evolution, successful parasite survival is
contingent on its own evolutionary adaptation to both of
them. To adapt, the parasite must use host signals and even
host immune responses to achieve parasitism. The immune
responses of the vertebrate host during malaria infection are
complex and not well understood, although different animal
models provide insight into possible effector mechanisms.
In the light of the complexity of Plasmodium development,
different immune response components evidently operate
at different stages of its life cycle in different organs, but little
is known about their fine specificity, dynamics, regulation
and/or influence on infection outcome. While some of these
responses are protective, others help the parasite to evade
those measures or induce pathology. The infection induces
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as IFNγ, TNFα and IL-12 production during P. yoelii
sporozoites-induced infection in B6 mice. We found that the
early immune response to sporozoite-induced malaria is
characterize by a peak of IFNγ in the serum at day 5 of
infection produced by splenic CD4 T cells. We also observed
that IFNγ deficient mice develop less parasitemia than wild
type. Altogether these results challenge the current view
regarding the role of IFNγ in the immune response to
Plasmodium. An early IFNγ production during malaria can
be deleterious for the host. They data also highlight the
complex regulation of the primary immune response to
malaria (Soulard, 2009).
identification of five strains highly resistant to CM, whereas
most laboratory mouse strains are susceptible. Using a
genome-wide screening approach, we were able to link the
phenotype of CM resistance to marker loci located in two
different chromosomal regions Berr 1 (chromosome 1) and
Berr 2 (chromosome 11), using a backcross cohort derived
from the CM resistant strains (WLA) and the C57BL/6
laboratory strain (CM susceptible). On the other hand, the
analysis of F2 mice derived from the same combination (WLA
and C57BL/6) revealed three disease phenotypes. Two of
them, CMSHPS (B6 phenotype) and CMRHPS (WLA
phenotype) were expected, and a third new phenotype,
CMRHPR, emerged. The animals displaying the latter are able
to control their parasitemia efficiently. This resistance
phenotype has not been reported previously and offers a
promising experimental model of chronic malaria that
appears more relevant to the human disease than the animal
models currently used. We have also obtained evidence for
linkage of this HP resistance phenotype to Berr 1 and to
another marker locus on chromosome 9 (Berr 3). Congenic
mouse strains for the chromosomal regions so far identified
respectively associated with CM and HP resistance are under
construction.
1.2 Immune responses associated with CM pathology
induced by P. berghei ANKA.
Cerebral Malaria (CM) occurs in 10-15% of P. falciparum
infected patients and is responsible for high mortality. CM is
characterized by 2 main factors: sequestration of parasitized
erythrocytes and the T-cell response. To date, the exact
mechanisms causing CM have not yet been clearly
established, but compelling evidence implicates T
lymphocytes in its development. Animal models, even if they
do not reproduce all the features of human CM, enabled us
to directly demonstrate T-cell involvement. Our findings
linked CM development to αβ T cells in mice infected with
P. berghei ANKA, a strain able to induce CM in susceptible
mice. Thus, we characterized the αβT-cells subsets and
studied their pathogenic functions. We demonstrated that
CM development in P. berghei ANKA -infected mice was
correlated with a significantly higher number of
CD8+TCRVβ8+ cells in peripheral blood (PBL) and in the
brains of mice that had been infected with sporozoites or
blood stage parasites than mice not developing CM (CM–).
Phenotypically, these CD8+ T cells were activated and they
secreted IFNγ and TNFα, two cytokines known to be
implicated in CM. In spite of all these indications, the exact
role played by these CD8+ T cells is still unknown. In
addition, the mechanisms involved in the control of the
balance between the efficient immune responses that
control parasite infection and inefficient responses leading
to chronic infections or pathological manifestations are
unknown. As a result, we consider that one of the
mechanisms could be an inefficient control of P. berghei
ANKA-induced pathogenic effector T cells by regulatory T
cells (Treg). Therefore, we evaluated the role of Treg cells in
CM in P. berghei ANKA infected B6 mice. We found more
activated CD4+CD25high T cells expressing Foxp3, with a
biased CDR3 TCRVß repertoire and in vitro suppressive
function during the course of the infection. However, in vivo
those cells do not protect against CM, because CM was not
exacerbated in mice depleted of their CD25+ Treg cells.
Moreover, this absence of protection was not due to in vivo
blockade of the Treg cells suppressive function by interleukin
(IL)-6 produced during the infection, because IL-6–depleted
or –deficient B6 mice were as susceptible as the controls to
CM (Vigário AM.; 2007).
1.4 Studies on Pf-infected patients.
To elucidate the role of immune response in the pathogenesis
of P. falciparum infection and particularly in CM, a study was
started on 2 cohorts of patients from different geographic
areas: 1) Gondia, India, an endemic village in northeastern
Maharashtra state where transmission is seasonal
(collaboration Dr. Gyan C. Mishra, NCCS, Pune, India, project
funded by IFCPAR) and 2) Libreville, Gabon, where
transmission is holo-endemic (collaboration Pr. Maryvonne
Kombila, Department of Parasitology-Mycology, Libreville
Hospital Center, Gabon, PAL+ project funded). The Gabonese
cohorts are constituted of 310 children (140 girls and 170
boys) of 2 months to 5 years-old. The Indian cohorts consist
of approximately 100 adult patients, including endemic and
non–endemic uninfected controls. P. falciparum infected
individuals were divided into five groups according to World
Health Organization criteria: 1) asymptomatic (AM), 2)
uncomplicated malaria (UM), 3) severe non-cerebral malaria
(SNCM), 4) CM and 5) individuals that recovered from CM (ex
CM). After obtaining informed consent (patients or their
parents or guardians) and Ethics Committee approval, blood
samples were collected at hospital admission (day 0), then on
days 7 and 30 after treatment.
Due to the multi-factorial character of malaria, we used an
integrated approach that addresses numerous interconnected
aspects of transmission, infection, immune responses and
disease by studying 1) T cell populations looking at their
phenotypes and repertoire diversity; 2) global profiles of
autoreactive antibodies to brain proteins produced during
infection using the PANAMA-BLOT method, 3) Profile of P.
falciparum specific antibodies, 4) P. falciparum genotypings
and 5) patterns of circulating cytokines.
Several observations have already been made from these
studies. In particular, a correlation between an increase of
self-reacting antibodies with disease severity. It is well
estabkshed that hypergammaglobulinemia and polyclonal
B-cell activation commonly occur in malaria, a fraction of
antibodies produced recognize self-components from
various tissues and organs. However, it is unclear whether
1.3 Identification of genetic traits controlling host resistance
to CM.
Host genes are among the main determinants of CM
susceptibility/resistance. Initial studies conducted using 12
inbred mouse strains (derived from ancestor pairs trapped in
different localities in Europe and North Africa) led to the
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these autoantibodies play a role in protective immunity or
contribute to the events leading to development of
neurological complications. We found that CM patients
develop a high autoantibody response to two brain
autoantigens, the alpha-2-spectrin (Guiyedi, 2007) and the
beta-tubulin-3 (Bansal, 2009). These autoantibody
reactivities were correlated positively with increased plasma
concentrations of cytokines that have been previously
associated with CM. These data strongly suggest the
involvement of an autoimmune counterpart targeting brain
antigens in CM. Clearly, the role of autoreactive antibodies
in the pathogenesis of CM, and other severe malaria
manifestations should be investigated further.
In conclusion, this multiparametric analytical approach,
combined with multivariate statistical analyses (MANOVA,
discriminant analysis, k-mean clustering) should help us to
elucidate the role of the immune response in the outcome of
P. falciparum infection. We hope that this strategy will allow
us to define the signatures of the various components of the
immune response associated with clinical subphenotypes of
P. falciparum malaria.
and the amplitude of the induced immune response, and
this action was effective when these specific motifs were
functionally recognized by Toll-like receptor 9 (TLR9).
2.2 Immuno-epidemiological studies on schistosomiasis
and malaria
We studied acquired immunological reactions during
schistosomiasis in patients from endemic areas. These
epidemiological studies have been completed by a
programme focusing on the changes in the immune
responses during co-infection, associating schistosomiasis
and malaria. In this way, we started to develop research on
the causes of variability of the host response during parasite
infection and their consequences on pathogenesis. These
studies shed light on the immune response acquired during
natural infection and helped us to design protocols of clinical
trials adapted to the selected target population. Recently, in
collaboration with UR024 (F. Simondon & F. Remoué) and
UR016 (D. Fontenille) of IRD, the laboratory of vector and
parasite ecology of Dakar University (UCAD; L. Konaté), the
medical entomology laboratory of Pasteur Institute of Dakar
(I. Dia), and the parasite biology and epidemiology
laboratory of IMTSSA (Ch. Rogier, Marseille), we developed
immunological studies in the field on the relationships
between the human host and vectors in the context of
malaria infections. We determined the influence of
environmental heterogeneities on vector bionomics and
malaria epidemiology in the Senegal River basin, and the
variability of the antibody response to P. falciparum in
children according to their exposure of Anopheles gambiae
s.l or Anopheles funestus vectors.
2. Clinical Immunology of Malaria
and Schistosomiasis
Since 1992, our group has developed two different
approaches on immunity against schistosomiasis, i) laboratory
research devoted to vaccine development using the
schistosome 28-kDa glutathione S-transferase (28GST), and ii)
studies on human acquired immunity in the endemic region
of the Senegal River. Since the region, like numerous subsaharian regions, is endemic for malaria, research in the field
has been extended to studies on acquired immunity during
co-infection (schistosomiasis and malaria). Today, our research
is oriented to the clinical development of our schistosomiasis
vaccine candidate, and to various environmental factors
influencing the immune response against malaria infection
and its pathology, including co-infection, nutritional
environment and vector density.
2.3 Clinical development of schistosomiasis vaccine
Progress has been made to limit the disease severity by using
chemotherapy, but continuous re-infection and risks of drug
resistance point to the necessary development of alternative
strategies. It is widely agreed that immunological prevention
of chronic parasitic infections will be extremely difficult to
achieve. Conversely, in some major helminth infections like
schistosomiasis, where parasite egg laying in the tissues is
the exclusive cause of pathology, inhibition of parasite
fecundity might represent for the future a novel way to
prevent the deleterious effects of these chronic infections in
man. The concept to target by vaccination the cause of the
pathology (fecundity) rather than the parasite itself has been
developed by our laboratory. Our work has designated
28GST as a potent candidate for therapeutic vaccination, and
should provide a potent tool to control a major chronic
infection. In the frame of the clinical development of our
vaccine candidate against schistosomiasis, the 28-kDa GST
of S. haematobium (Sh28GST), the team initiated, developed
and coordinated several clinical studies in humans. This
protein, selected as a therapeutic vaccine candidate, has
been produced under GMP conditions by Eurogentec S.A.,
and has been evaluated in pre-clinical toxicological studies.
Following a first safety phase undertaken in the CIC of Lille
and funded by Inserm, three clinical phases in Senegal and
one in Niger demonstrated that this candidate was safe and
immunogenic in infected children, which represent the
target population. These clinical studies were supported by
an EC demonstration project.
2.1 Immuno-pharmacology of vaccine
Vector design for mucosal or cutaneous vaccination against
schistosomiasis has been extensively developed. Synthetic
(liposomes, microparticules) and recombinant live vectors
(BCG, attenuated B. pertussis, attenuated S. thyphi) were
engineered in collaboration with several European
laboratories including the C. Locht team. All these vectors
were adapted for mucosal immunisation and provided a
unique panel of tools and models to develop future vaccine
processes. As an example, a single-dose mucosal
administration with biodegradable microparticles produced
by spray-drying technique with particular polyester
polymers and entrapping Schistosoma mansoni antigen,
was enough to generate protective immune response in a
mouse model. A nucleic acid vaccine was also developed
using the 28GST gene. Beside the fact that this kind of
vaccine presentation could be very effective in experimental
models, we determined that the constitution of the DNA
vector is a crucial element, which could influence the quality
of the immune response and could be adapted to generate
appropriate effects. We showed that a methylated CpGcontaining plasmid has a particular role in the orientation
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Poulain-Godefroy O, Vendeville C, Locht C, Riveau, G.
Bordetella pertussis filamentous haemagglutinin delivered by mucosal
routes enhances immunoglobulin level in serum and mucosal fluids
FEMS.
Immunol Med Microbiol, 2008, Oct;54:129-136.
development 2010
2009
We will pursue the projects that have shown the most
potential and promise. Basic and field studies will be
conducted in parallel, often in a highly interactive manner to
study the additive role of environment and host-parasite
interactions in malaria and in its degrees of severity. We will 1)
evaluate the appropriateness of the immune response during
malaria, 2) identify biomarkers associated to cerebral malaria,
3) identify host genetic factors associated with protection or
pathological processes and 4) characterize environmental
factors that modulate these responses.
Understanding the duality of immune responses to malaria is
expected to contribute to the design of new therapeutics and
vaccine strategies.
Bansal D, Herbert F, Lim P, Deshpande P, Bécavin C, Guiyedi V, de
Maria I, Rousselle JC, Namane A, Jain R, Cazenave PA, Mishra GC,
Ferlini C, Fesel C, Benecke A, Pied S.
IgG autoantibody to brain beta tubulin III associated with cytokine
cluster-II discriminate cerebral malaria in central India.
PLoS One, 2009, 4:e8245.
Driss V, Legrand F, Hermann E, Loiseau S, Guerardel Y, Kremer L,
Adam E, Woerly G, Dombrowicz D, Capron M.
TLR2-dependent eosinophil interactions with mycobacteria : role of
alpha-defensins.
Blood, 2009, 113:3235-3244.
Fillol F, Sarr JB, Boulanger D, Cisse B, Sokhna C, Riveau G, Simondon
KB, Remoué F.
Impact of child malnutrition on the specific anti-Plasmodium falciparum
antibody response.
Malar J, 2009, 8:116.
Publications
Guiyedi V, Deshpande P, Fesel C, Jain R, Dzeing-Ella A, Cazenave P-A,
Kombila M, Mishra G C, Pied S.
Correlation of Plasma Soluble Fas Ligand Levels with Severe Anaemia in
Gabonese and Indian Plasmodium falciparum Malaria Patients.
The Open Trop Med J, 2009, 2:17-23.
2007
Duarte J, Deshpande P, Guiyedi V, Mecheri S, Fesel C, Cazenave PA,
Mishra GC, Kombila M, Pied S.
Total and functional parasite specific IgE responses in Plasmodium
falciparum-infected patients exhibiting different clinical status.
Malar J, 2007, 6:1
Legrand F, Driss V, Woerly G, Loiseau S, Hermann E, Fournié JJ,
Héliot L, Mattot V, Soncin F, Gougeon ML, Dombrowicz D, Capron M.
A functional gammadeltaTCR/CD3 complex distinct from gammadeltaT
cells is expressed by human eosinophils.
PLoS One, 2009, 4:e5926.
Guiyedi V, Chanseaud Y, Fesel C, Snounou G, Rousselle JC, Lim P,
Koko J, Namane A, Cazenave PA, Kombila M, Pied S.
Self-reactivities to the non-erythroid alpha spectrin correlate with
cerebral malaria in Gabonese children. 2007.
PLosOne, 2007, 2:e389.
Soulard V, Roland J, Gorgette O, Barbier E, Cazenave PA, Pied S.
An early burst of IFN-gamma induced by the pre-erythrocytic stage
favours Plasmodium yoelii parasitaemia in B6 mice.
Malaria J, 2009, 8:128.
Sarr JB, Remoue FJ, Samb B, Dia I, Guindo SK, Sow C, Maiga S,
Tine S, Thiam C, Schacht AM, Simondon ., Konate L, Riveau G.
Evaluation of antibody response to Plasmodium falciparum in children
according to exposure of Anopheles gambiae or Anopheles funestus vectors.
Malaria J, 2007, 6:117-125.
2010
Baptista FG, Pamplona A, Pena AC, Mota MM, Pied S, Vigário AM.
Accumulation of Plasmodium-infected red blood cells in the 1 brain is
crucial for the 2 development of cerebral malaria in mice.
Infect Immunity, 2010, 78:4033–4039.
Soulard V, Roland J, Sellier C, Gruner AC, Leite-De-Moraes M,
Franetich JF, Renia L, Cazenave PA, Pied S.
Primary infection of C57BL/6 mice with Plasmodium yoelii induces a
heterogeneous response of NKT cells.
Infect Immun, 2007, 75:25112522.
Diallo TO, Remoue F, Gaayeb L, Schacht AM, Charrier N, De Clerck
D, Dompnier JP, Pillet S, Garraud O, N'diaye A, Riveau G.
Schistosomiasis coinfection in children influences acquired immune
response against Plasmodium falciparum malaria antigens.
PLoSONE, 2010, 5:e12764.
Vigário AM, Gorgette O, Dujardin HC, Cruz T, Cazenave PA,
Adrien Six, Bandeira A, Pied S.
Regulatory CD4+ T cells expand during experimental Plasmodium
infection but do not prevent cerebral malaria.
Int J Parasitol, 2007, 37:963-973.
Diop MA, Riveau G, Tendeng L, Sallet G. A METAPOPULATION.
Model for Transmission of Schistosomiasis.
Mathematical Biosciences, 2010, In press.
2008
Poinsignon A, Samb B, Doucoure S, Drame PM, Sarr JB, Sow C,
Cornelie S, Maiga S, Thiam C, Rogerie F, Guindo S, Hermann E,
Simondon F, Dia I, Riveau G, Konate L, Remoue F.
A First attempt to validate the gSG6-P1 salivary peptide as an imunoepidemiological tool for evaluating human exposure to An. funestus bites.
Trop Med & Int Health, 2010, 15:1198-1203.
Bompart F, Hirsch F, Bertoye PH, Vray M, Riveau G. participants in
Round Table No1, Giens XXIII.
Good clinical practice in developing countries : applying recommendations.
Therapie, 2008, 63:83-8, 77-82.
Dia I, Konate L, Samb B, Sarr JB, Diop A, Rogerie F, Faye M,
Riveau G, Remoue F, Diallo M, Fontenille D.
Bionomics of malaria vectors and relationship with malaria transmission
and epidemiology in three physiographic zones in the Senegal River Basin.
Acta Tropica, 2008, 105:145-153.
58
Contents
Pulmonary immunity
1. Endothelium and pulmonary
inflammation
Anne TSICOPOULOS, DR2 Inserm
(P. Lassalle and C. Glineur)
Group Members :
1.1 Discovery of the functions of a new pulmonary
endothelial proteoglycan (PG) : endocan.
Role of endocan in sepsi : endocan, a lung EC PG, the cDNA
of which was cloned in our lab, is a modulator of the integrin
LFA-1 and may modulate leukocyte recruitment. The normal
blood endocan level is around 1 ng/mL. In patients with
severe sepsis, blood levels of endocan increase from 5 to 100
fold. To evaluate the role of endocan in animal models,
recombinant mouse endocan was produced, and a specific
mouse/rat ELISA was developed. Endotoxinic rats treated with
the anti-protease calpain showed a reduction of leukocyte
rolling and adhesion to the mesenteric vessels, a reduction in
cardiac lesions, and increased serum endocan levels,
suggesting a protective role of endocan in sepsis.
Lassalle Philippe, CR1 Inserm
Duez Catherine, CR1 Inserm
Glineur Corine, CR1 CNRS
Delehedde Maryse, DR contractuel Inserm
Scherpereel Arnaud, PU- PH Univ Lille 2/CHRU
Tonnel André-Bernard, PU-PH Univ Lille 2/CHRU
Wallaert Benoît, PU-PH Univ Lille 2/CHRU
Perez Thierry, PH Univ Lille 2/CHRU
Ramon Philippe, PH Univ Lille 2/CHRU
Just Nicolas, PH Univ Lille 2/CHRoubaix
Moreau Caroline, PH Univ Lille 2/CHRU
Devos David, MCU Univ Lille 2/CHRU
Vorng Han, CE1 IPL
Hauw Philippe, Technician IPL
Kervoaze Gwenola, Technician Inserm
Marchandise Geneviève, Technician IPL
Marquillies Philippe, Technician IPL
De Freitas Caires Nathalie, Postdoctoral fellow Inserm
Ait Yahia Saliha, PhD Student Univ Lille 2
Azzaoui Imane, PhD Student Univ Lille 2
Bousquet Guilhem, PhD Student, Paris St Louis
Chaouche Siham, PhD Student, ENS Alger, Algerie
Ple Coline, PhD Student Univ Lille 2
Awad Ali, Master 2 Student Univ Lille 2
Pastre Jean, Master 2 Student Univ Lille 2
Role of endocan in cancer : Endocan has been defined as a
tumor endothelial marker in non small cell lung cancer (NSCLC)
and in hepatocarcinoma. Survival inversely correlated with
endocan blood levels in NSCLC, and with microvascular density
measured by endocan in hepatocarcinoma. Endocantransfected HEK cells formed tumors when injected in SCID
mice. Both the glycan and the protein cores were involved in
endocan tumorigenesis. Thus our working hypothesis is that
endocan, by inhibiting LFA-1-dependent transendothelial cell
migration of NK cells (see preliminary experiments in part 1.1
of the project), may prevent tumor leukocyte infiltration. We
also found that mesothelin, which is a 40-kDa glycoprotein
shed from the mesothelial cell surface, was increased in blood
and pleural fluid from malignant pleural mesothelioma (MPM),
and constituted a bad prognosis marker of MPM. Overall, these
data suggest that endocan may be used as a novel endothelial
dysfunction marker in inflammatory pulmonary diseases and
may represent a therapeutic target in lung cancer.
Key words :
Respiratory diseases. Allergy. Asthma. Sepsis. Lung Cancer.
COPD. Endothelial cells. Proteoglycans. NK cells. Chemokines.
Adaptive immunity. Animal models of respiratory diseases.
A - Research Report
1.2 Nature of the interactions between proteoglycans and
inflammatory mediators.
Cyclophilin B is an inflammatory mediator able to bind
cyclosporin A, which mediates the chemotaxis of memory T
cells by binding to PGs. The specific sequence of interaction
was identified as 3-O-sulfated N-unsubstituted glucosamine
residue (collaboration Fabrice Allain, Lille). In contrast,
hepatocyte growth factor/scatter factor (HGF/SF) required
mainly electrostatic interactions through iduronate residues.
HGF/SF also interacted with other molecules than PGs, such
as Neuropilin-1, through a “heparin” mimetic site, which
modulated its activities on endothelial cells. Therefore,
complex interactions taking place between endothelial
proteoglycans and mediators may modify the course of the
inflammatory reaction.
Among respiratory diseases, allergic asthma, pulmonary
infections and lung cancer represent major problems of public
health. These diseases affect millions of people, are in constant
increase and are a major cause of morbidity and mortality.
Although considerable therapeutic progress has been made
over the last 20 years, there is still no treatment able to modify
the natural course of chronic respiratory diseases. These
diseases share common key target cells involved in their
pathogenesis: the epithelial cells as the first line of defence,
the inflammatory cells as one of the early host-response
events, and the endothelial cells (EC) as a link between
epithelial and inflammatory cells. Our goals have been to
evaluate how these cells and their mediators can orchestrate
the host inflammatory reaction and tissue remodeling in
response to allergens, bacteria or tumor cells.
1.3 Neurodegenerative disorders, endothelium and
hypoxia
Amyotrophic lateral sclerosis (ALS) is a progressive motor
neuron degenerative disease, in which the prognosis is
governed by a progressive respiratory failure. Animal models
have suggested that an abnormal response of VEGF to hypoxia
may be at the origin of the ALS. We have shown that ALS
patients indeed exhibited a hypo-responsiveness to hypoxia
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Contents
specific to VEGF and to the disease. Thus abnormal angiogenic
responses to hypoxia may lead to neurodegenerative and
lung disorders.
of BCG or BCG-IL-18 before OVA-sensitization prevented Th2
cytokine production, eosinophilic inflammation and AHR.
Intratracheal instillation of BCG-IL-18, once the pulmonary
allergic reaction was established, also inhibited the asthmatic
reaction, with a concomitantly increased number of NK cells.
This prompted us to evaluate NK cells in the regulation of
allergic asthma (see part 3 of the project). Altogether these
data suggest that bacteria can redirect the deleterious Th2
response, and that innate cells such as NK cells may play a role
in this effect.
2. Cell homing and pulmonary
inflammation (Leaders : A Tsicopoulos and C Duez)
2.1 New insights into the mechanisms involved in allergic
asthma.
DC are usually considered as key players in the initiation of the
allergic reaction. In a murine model of ovalbumin (OVA)-driven
asthma, it was shown that the depletion of DC during allergen
challenges abolished the characteristic features of asthma,
which were restored by adoptive transfer of DC, (collaboration
BN Lambrecht, Rotterdam, The Netherlands). Thus, DC also have
a key role in ongoing asthma. Eosinophils are considered as
effector cells in asthma. However, following intranasal
administration of OVA, eosinophils expressing high levels of
MHC class II and CD86 were found in the mediastinal lymph
nodes (MLN), suggesting that they may have an antigenpresenting activity in ML (Collaboration EW Gelfand, Denver,
USA). The relationship between neutrophils and airway
hyperresponsiveness (AHR) remains unclear. Following allergen
challenge, we observed an early and transient neutrophil
infiltration mediated by IL-1 and IL-18. The blockade of IL-1 and
IL-18 did not affect the development of AHR and eosinophilic
airway inflammation, thereby suggesting that neutrophils do
not play a direct role in allergen induced AHR.
2.3 Establishment of original humanized SCID mouse
models mimicking human diseases.
The absence of animal models able to accurately reproduce
human diseases led us to set up a humanized SCID mouse
model consisting in double grafts of human skin and
peripheral mononuclear cells. Using this model, it was shown
that cutaneous CCL17 can translocate to the skin draining
lymph nodes and mediate the recruitment of human memory
CD4+ cells and DC in lymph nodes, followed by a
redistribution of Th2 cells but not Th1 cells in the skin. These
data argue for a role of CCL17 in both the initiation and the
development of Th2-associated skin inflammation.
2.4 Breakthroughs in clinical studies
Clinical research involves several departments of the regional
university hospital of Lille, but mostly the pulmonary
department, and has been devoted to the search for new
diagnosis tools, new prognosis biomarkers (see part 2.2), and
novel therapeutic approaches in pulmonary diseases. In
particular, the beneficial effect of acetylcysteine in idiopathic
pulmonary fibrosis or the anti-inflammatory effect of opiate
derivatives on mucosal explants from inflammatory bowel
diseases was observed (collaboration M Chamaillard,
U795/CIIL). Overall, these results highlight the translation of
research from bench to bedside.
The chemokine CCL18 is preferentially expressed in the lung,
is induced by Th2 cytokines and attracts naive T cells and DC.
We showed that allergen stimulation induced the production
of CCL18 by monocytes from asthmatics through an IL-4/IL-13
dependent pathway. CCL18 also attracted polarized Th2 cells
and basophils, and was increased in broncho-alveolar lavage
fluids from allergic asthmatics. However, CCL18 is also induced
by IL-10, a suppressive cytokine, and inhibits eosinophil
recruitment. Ongoing work shows that under physiological
conditions, CCL18 recruits regulatory T cells, and directly affect
T cells and DC in favour of a regulatory profile. Altogether
these results show an unusual anti-inflammatory effect of a
chemokine under physiological conditions, which seems to
be dysregulated in allergic asthma.
2.2 Advances in the regulation of allergic asthma by
environmental factors.
Pollutants are a major environmental factor promoting allergic
asthma. We and others have previously shown that diesel
exhausts exacerbate cellular inflammation in already
sensitized patients. Diesel exposure of mononuclear cells from
nonatopic donors induced an increased production of CCL18,
and a decreased production of CXCL10 (a pro-Th1
chemokine), leading to the recruitment of Th2 cells. Therefore,
diesel exposure may be involved in the increased prevalence
of allergic diseases.
Bacteria can also regulate allergic diseases. In an in vitro
polarized intestinal epithelium model, probiotic bacteria
activated DC from allergic patients, inducing IFN-γ production
by T cells, (collaboration C Grangette, CIIL), and inhibited DCinduced Th2 responses, even after re-challenge with the
allergen alone. BCG has a Th1 immunostimulatory activity. To
increase this effect, a BCG producing IL-18 (BCG-IL-18) was
constructed (collaboration F Biet, C Locht, CIIL). Administration
Perspectives and
development 2010
The goal of our team will be to characterize some of the
mechanisms involved in the pulmonary immune response
associated with these respiratory diseases in order to highlight
potential therapeutic strategies. Three themes will be
developed:
Theme 1: Endothelium response to environmental factors
(inflammatory stress, infection and hypoxia): Endothelial cells
are on the front line to regulate leukocyte influx to
inflammatory sites. They produce proteoglycans (PGs) which
may affect the distribution and functions of mediators
associated to immune inflammatory responses. We will study
the control of leukocyte infiltration and activation by endocan
(PG preferentially expressed by lung endothelial cells) and
endocan peptide fragments, and analyze its role in asthma
and COPD. Moreover, the endothelial vascular response to
environmental factors will be characterized by identifying its
PG profile and its functional consequences. This project should
allow to decipher the functional role of endothelial PGs, which
60
Contents
Dusser D, Montani D, Chanez P, de Blic J, Delacourt C, Deschildre A,
Devillier P, Didier A, Leroyer C, Marguet C, Martinat Y, Piquet J,
Raherison C, Serrier P, Tillie-Leblond I, Tonnel AB, Tunon de Lara M,
Humbert M.
Mild asthma: an expert review on epidemiology, clinical characteristics
and treatment recommendations.
Allergy, 2007, 62:591-604.
may represent key components of vascular remodeling,
providing new targets for follow-up and therapeutic
intervention in asthma and COPD.
Theme 2: Control of the pulmonary immune response by
chemokines: Chemokines and their receptors actively
participate to the inflammatory reaction in the lung not only
by the recruitment of effector and regulatory cells, but also by
their direct remodeling and immunoregulatory effects. The
project will first focus on CCL18, which we showed to be
associated with asthma. The regulation of the pulmonary
adaptive immune response by CCL18 through T cells and
dendritic cells and in experimental asthma will be studied. The
characterization of its receptor aims to identify a homing
chemokine receptor targeting the lung. Finally, to open news
perspectives on the regulatory functions of chemokines, their
contribution to the pulmonary immune response to NOD-like
receptor (NOD)1 in asthma will be determined (collaboration
M Chamaillard). These studies should allow to unravel the
complex in vivo role of CCL18 in asthma, and open new
insights in the links between innate and adaptive immunity
in asthma.
Theme 3: Regulation of pulmonary immunity by Natural Killer
(NK) cells: NK cells are sensors of the environment bridging
innate and adaptive immunity. Although present in high
numbers in the lungs, their recruitment and homing to this
organ and their role in the immune response associated with
asthma are largely unknown. The project aims to identify the
role of NK cells in the regulation of acute and chronic
experimental asthma, NK cell subsets or activation pathways
involved in this regulation, and factors involved in their
recruitment to the lungs and mediastinal lymph nodes. Their
implication in the regulation of experimental asthma induced
by Toll-like receptor2 and NOD2 will also be determined.
Overall, this work should help to characterize the role of NK
cells in the regulation of pulmonary immunity in asthma, and
may allow identification of new therapeutic targets.
Altogether, this project should identify novel therapeutic
strategies in asthma and COPD.
Freitag L, Ernst A, Unger M, Kovitz K, Marquette CH.
A proposed classification system of central airway stenosis.
Eur Respir J, 2007, 30:7-12.
Greillier L, Cavailles A, Fraticelli A, Scherpereel A, Barlesi F, Tassi G,
Thomas P, Astoul P.
Accuracy of pleural biopsy using thoracoscopy for the diagnosis of
histologic subtype in patients with malignant pleural mesothelioma.
Cancer, 2007, 110:2248-2252.
Grigoriu BD, Scherpereel A, Devos P, Chahine B, Letourneux M,
Lebailly P, Grégoire M, Porte H, Copin MC, Lassalle P.
Utility of osteopontin and serum mesothelin in malignant pleural
mesothelioma diagnosis and prognosis assessment.
Clin Cancer Res, 2007, 13:2928-2935.
Janssen JP, Collier G, Astoul P, Tassi GF, Noppen M, RodriguezPanadero F, Loddenkemper R, Herth FJ, Gasparini S, Marquette CH,
Becke B, Froudarakis ME, Driesen P, Bolliger CT, Tschopp JM.
Safety of pleurodesis with talc poudrage in malignant pleural effusion :
a prospective cohort study.
Lancet, 2007, 369:1535-1539.
Just N, Moreau C, Lassalle P, Gosset P, Perez T, Brunaud-Danel V,
Wallaert B, Destée A, Defebvre L, Tonnel AB, Devos D.
High erythropoietin and low vascular endothelial growth factor levels in
cerebrospinal fluid from hypoxemic ALS patients suggest an abnormal
response to hypoxia.
Neuromuscul Disord, 2007, 17:169-73.
Makris D, Hatthabi M, Scherpereel A, Lafitte JJ, Marquette CH.
Haemothorax after pig-tail catheter removal in a patient with primary
spontaneous pneumothorax.
Emerg Med J, 2007, 24:e17.
Makris D, Marquette CH.
Tracheobronchial stenting and central airway replacement.
Publications
Makris D, Scherpereel A, Copin MC, Colin G, Brun L, Lafitte JJ,
Marquette CH.
Fatal interstitial lung disease associated with oral erlotinib therapy for
lung cancer.
BMC Cancer, 2007, 7:150.
2007
Does not Increase Allergic Airway Disease in Mice When Produced by BCG.
J Biomed Biotechnol, 2007:67276.
Makris D, Scherpereel A, Leroy S, Bouchindhomme B, Faivre JB,
Remy J, Ramon P, Marquette CH.
Electromagnetic navigation diagnostic bronchoscopy for small
peripheral lung lesions.
Eur Respir J, 2007, 29:1187-1192.
Berghmans T, Lafitte JJ, Lecomte J, Alexopoulos CG, Van Cutsem O,
Giner V, Efremidis A, Berchier MC, Collon T, Meert AP,
Scherpereel A, Ninane V, Leclercq N, Paesmans M, Sculier JP ;
European Lung Cancer Working Party.
Second-line paclitaxel in non-small cell lung cancer initially treated with
cisplatin: a study by the European Lung Cancer Working Party.
Br J Cancer, 2007, 96:1644-1649.
Nseir S, Duguet A, Copin MC, De Jonckheere J, Zhang M,
Similowski T, Marquette CH.
Continuous control of endotracheal cuff pressure and tracheal wall
damage: a randomized controlled animal study.
Crit Care, 2007, 11:R109.
Collet F, Mallart A, Bervar JF, Bautin N, Matran R, Pattou F,
Romon M, Perez T.
Physiologic correlates of dyspnea in patients with morbid obesity.
Int J Obes (Lond), 2007, 31:700-706.
Ratajczak C, Duez C, Grangette C, Pochard P, Tonnel AB, Pestel J.
Impact of lactic Acid bacteria on dendritic cells from allergic patients in
an experimental model of intestinal epithelium.
J Biomed Biotechnol, 2007, 2007:71921.
Conti M, Robin E, Porte H, Marquette CH, Wurtz A.
Management of postintubation tracheobronchial ruptures.
Ann Thorac Surg, 2007, 83:1924.
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Contents
Berghmans T, Gourcerol D, Lafitte JJ, Kotsori K, Paesmans M,
Scherpereel A, Leclercq N, Sculier JP; for the European Lung Cancer
Working Party.
Mitomycin plus vinorelbine salvage chemotherapy in non-small cell lung
cancer : a prospective study.
Lung Cancer, 2008, 61:378-384.
Rennel E, Mellberg S, Dimberg A, Petersson L, Botling J, Ameur A,
Westholm JO, Komorowski J, Lassalle P, Cross MJ, Gerwins P.
Endocan is a VEGF-A and PI3K regulated gene with increased expression
in human renal cancer.
Exp Cell Res, 2007, 313:1285-1294.
Scherpereel A; French Speaking Society for Chest Medicine (SPLF)
Experts Group.
Guidelines of the French Speaking Society for Chest Medicine for
management of malignant pleural mesothelioma.
Respir Med, 2007, 101:1265-1276.
Carré PC, Roche N, Neukirch F, Radeau T, Perez T, Terrioux P,
Ostinelli J, Pouchain D, Huchon G.
The Effect of an Information Leaflet upon Knowledge and Awareness of
COPD in Potential Sufferers. A Randomized Controlled Study.
Respiration, 2008, 76:53-60.
Scherpereel A, Lee YC.
Biomarkers for mesothelioma.
Catlow KR, Deakin JA, Wei Z, Delehedde M, Fernig DG, Gherardi E,
Gallagher JT, Pavão MS, Lyon M.
Interactions of Hepatocyte Growth Factor/Scatter Factor with Various
Glycosaminoglycans Reveal an Important Interplay between the Presence
of Iduronate and Sulfate Density.
J Biol Chem, 2008, 283:5235-5248.
Sculier JP, Lafitte JJ, Lecomte J, Alexopoulos CG, Van Cutsem O,
Giner V, Efremidis A, Berchier MC, Collon T, Meert AP, Scherpereel A,
Ninane V, Paesmans M, Berghmans T ; European Lung Cancer
Working Party.
A phase III randomised trial comparing sequential chemotherapy using
cisplatin-based regimen and paclitaxel to cisplatin-based chemotherapy
alone in advanced non-small-cell lung cancer.
Ann Oncol, 2007, 18:1037-1042.
Depontieu F, Grigoriu BD, Scherpereel A, Adam E, Delehedde M,
Gosset P, Lassalle P.
Loss of Endocan tumorigenic properties after alternative splicing of exon 2.
BMC Cancer, 2008, 8:14.
Sculier JP, Lafitte JJ, Paesmans M, Lecomte J, Alexopoulos CG,
Van Cutsem O, Giner V, Efremidis A, Berchier MC, Collon T, Meert AP,
Scherpereel A, Ninane V, Koumakis G, Vaslamatzis MM, Leclercq N,
Berghmans T ; European Lung Cancer Working Party.
Chemotherapy improves low performance status lung cancer patients.
Eur Respir J, 2007, 30:1186-1192.
Ferrari F, Liu ZH, Lu Q, Becquemin MH, Louchahi K, Aymard G,
Marquette CH, Rouby JJ.
Comparison of lung tissue concentrations of nebulized ceftazidime in
ventilated piglets: ultrasonic versus vibrating plate nebulizers.
Intensive Care Med, 2008.
Grigoriu BD, Scherpereel A.
Diagnostic value of soluble mesothelin in malignant mesothelioma.
Thorax, 2008, 63:87-88.
Shuvaev VV, Christofidou-Solomidou M, Scherpereel A, Simone E,
Arguiri E, Tliba S, Pick J, Kennel S, Albelda SM, Muzykantov VR.
Factors modulating the delivery and effect of enzymatic cargo
conjugated with antibodies targeted to the pulmonary endothelium.
J Control Release, 2007, 118:235-244.
Plé C, Duez C.
Toll-like receptor expressing cells for antiallergy compound screening.
Expert Opinion on drug Discovery, 2008, 3:629-641.
Tillie-Leblond I, Gosset P, Le Berre R, Janin A, Prangère T, Tonnel AB,
Guery BP.
Keratinocyte growth factor improves alterations of lung perme>ability
and bronchial epithelium in allergic rats.
Eur Respir J, 2007, 30:31-39.
Revel MP, Faivre JB, Remy-Jardin M, Deken V, Duhamel A,
Marquette CH, Tacelli N, Bakai AM, Remy J.
Automated lobar quantification of emphysema in patients with severe
COPD.
Eur Radiol, 2008.
Tunon-de-Lara JM, Laurent F, Giraud V, Perez T, Aguilaniu B,
Meziane H, Basset-Merle A, Chanez P.
Air trapping in mild and moderate asthma : effect of inhaled corticosteroids.
J Allergy Clin Immunol, 2007, 119:583-590.
Roche N, Dalmay F, Perez T, Kuntz C, Vergnenègre A, Neukirch F,
Giordanella JP, Huchon G.
Impact of chronic airflow obstruction in a working population.
Eur Respir J, 2008, 31:1227-1233.
Vanpouille C, Deligny A, Delehedde M, Denys A, Melchior A,
Liénard X, Lyon M, Mazurier J, Fernig DG, Allain F.
The heparin/heparan sulfate sequence that interacts with cyclophilin B
contains a 3-O-sulfated N-unsubstituted glucosamine residue.
J Biol Chem, 2007, 282:24416-24429
Sapede C, Gauvrit A, Barbieux I, Padieu M, Cellerin L, Sagan C,
Scherpereel A, Dabouis G, Grégoire M.
Aberrant splicing and protease involvement in mesothelin release from
epithelioid mesothelioma cells.
Cancer Sci, 2008, 99:590-594.
2008
Adam E, Sarrazin S, Landolfi C, Motte V, Lortat-Jacob H, Lassalle P,
Delehedde M.
Efficient long-term and high-yielded production of a recombinant proteoglycan in eukaryotic HEK293 cells using a membrane-based bioreactor.
Tillie-Leblond I, Wislez M, Valeyre D, Crestani B, Rabbat A, IsraelBiet D, Humbert M, Couderc LJ, Wallaert B, Cadranel J.
Interstitial lung disease and anti-Jo-1 antibodies : difference between
acute and gradual onset.
Thorax, 2008, 63:53-59.
Berghmans T, Dusart M, Paesmans M, Hossein-Foucher C, Buvat I,
Castaigne C, Scherpereel A, Mascaux C, Moreau M, Roelandts M,
Alard S, Meert AP, Patz EF Jr, Lafitte JJ, Sculier JP; European Lung
Cancer Working Party for the IASLC Lung Cancer Staging Project.
Primary tumor standardized uptake value (SUVmax) measured on
fluorodeoxyglucose positron emission tomography (FDG-PET) is of
prognostic value for survival in non-small cell lung cancer (NSCLC) : a
systematic review and meta-analysis (MA) by the European Lung Cancer
Working Party for the IASLC Lung Cancer Staging Project.
J Thorac Oncol, 2008, 3:6-12.
Tsicopoulos A, Duez C, Saxon A.
Environmental Factors in IgE production. Allergy and Allergic Diseases
(Second Edition).
Blackwell publishing, 2008 Chapter 7
Tonnel AB, Pouwels-Frys S, Tillie-Leblond I.
Allergic bronchopulmonary aspergillosis. Allergy and Allergic Diseases
(Second Edition).
Blackwell publishing, 2008 Chapter 82
62
Contents
Maurage CA, Adam E, Minéo JF, Sarrazin S, Debunne M,
Siminski RM, Baroncini M, Lassalle P, Blond S, Delehedde M.
Endocan expression and localization in human glioblastomas.
J Neuropathol Exp Neurol, 2009, 68:633-641.
2009
Behr J, Demedts M, Buhl R, Costabel U, Dekhuijzen RP, Jansen HM,
MacNee W, Thomeer M, Wallaert B, Laurent F, Nicholson AG,
Verbeken EK, Verschakelen J, Flower CD, Petruzzelli S, De Vuyst P,
van den Bosch JM, Rodriguez-Becerra E, Lankhorst I, Sardina M,
Boissard G; IFIGENIA study group.
Lung function in idiopathic pulmonary fibrosis--extended analyses of the
IFIGENIA trial.
Respir Res, 2009, 10:101.
Moreau C, Gosset P, Brunaud-Danel V, Lassalle P, Degonne B,
Destee A, Defebvre L, Devos D.
CSF profiles of angiogenic and inflammatory factors depend on the
respiratory status of ALS patients.
Amyotroph Lateral Scler, 2009, 10:175-181.
Estrella C, Rocks N, Paulissen G, Quesada-Calvo F, Noël A, Vilain E,
Lassalle P, Tillie-Leblond I, Cataldo D, Gosset P.
Role of A disintegrin and metalloprotease-12 in neutrophil recruitment
induced by airway epithelium.
Am J Respir Cell Mol Biol, 2009, 41:449-458.
Perez T, Arnould B, Grosbois JM, Bosch V, Guillemin I, Bravo ML,
Brun M, Tonnel AB; TIPHON Study Group.
Validity, reliability, and responsiveness of a new short Visual Simplified
Respiratory Questionnaire (VSRQ) for health-related quality of life
assessment in chronic obstructive pulmonary disease.
Int J Chron Obstruct Pulmon Dis, 2009, 4:9-18.
Gilet J, Chang Ying, Chenivesse C, Legendre B, Vorng H, Duez C,
Wallaert B, Porte H, Senechal S, Tsicopoulos A.
Role of CCL17 in the generation of cutaneous inflammatory reactions in
hu-PBMC-SCID mice grafted with human skin.
J Invest Dermatol, 2009, 129:879-890.
Ryningen A, Stapnes C, Lassalle P, Corbascio M, Gjertsen BT,
Bruserud O.
A subset of patients with high-risk acute myelogenous leukemia shows
improved peripheral blood cell counts when treated with the
combination of valproic acid, theophylline and all-trans retinoic acid.
Leuk Res, 2009, 3:779-787.
Grigoriu B, Chahine B, Zerimech F, Grégoire M, Balduyck M,
Copin MC, Devos P, Lassalle P, Scherpereel A.
Serum mesothelin has a higher diagnostic utility than hyaluronic acid
in malignant mesothelioma.
Clin Biochem, 2009, 42:1046-50.
Roux AL, Catherinot E, Ripoll F, Soismier N, Macheras E, Ravilly S,
Bellis G, Vibet MA, Le Roux E, Lemonnier L, Gutierrez C, Vincent V,
Fauroux B, Rottman M, Guillemot D, Gaillard JL, Herrmann JL,
Wallaert B for the OMA Group.
Multicenter study of prevalence of nontuberculous mycobacteria in
patients with cystic fibrosis in france.
J Clin Microbiol, 2009, 47:4124-4128.
Grigoriu BD, Grigoriu C, Chahine B, Gey T, Scherpereel A.
Clinical utility of diagnostic markers for malignant pleural mesothelioma.
Monaldi Arch Chest Dis, 2009, 71:31-38.
Grigoriu B, Scherpereel A.
Screening for malignant mesothelioma--looking for the holy grail.
Am J Respir Crit Care Med, 2009, 179:851.
Taront S, Dieudonné A, Blanchard S, Jeannin P, Lassalle P,
Delneste Y, Gosset P.
Implication of scavenger receptors in the interactions between diesel
exhaust particles and immature or mature dendritic cells.
Part Fibre Toxicol, 2009, 6:9.
Hatfield K, Øyan AM, Ersvaer E, Kalland KH, Lassalle P, Gjertsen BT,
Bruserud Ø.
Primary human acute myeloid leukaemia cells increase the proliferation
of microvascular endothelial cells through the release of soluble
mediators.
Br J Haematol, 2009, 144:53-68.
Vandermeers F, Hubert P, Delvenne P, Mascaux C, Grigoriu B,
Burny A, Scherpereel A, Willems L.
Valproate, in combination with pemetrexed and cisplatin, provides
additional efficacy to the treatment of malignant mesothelioma.
Clin Cancer Res, 2009, 15:2818-2828.
Grigoriu BD, Chahine B, Vachani A, Gey T, Conti M, Sterman DH,
Marchandise G, Porte H, Albelda SM, Scherpereel A.
Kinetics of soluble mesothelin in patients with malignant pleural
mesothelioma during treatment.
2010
Barlési F, Balleyguier C, Besse B, Bonodeau F, Brenac F, Corneloup O,
Dansin E, Ferretti G, Gaubert JY, Gervais R, Lacombe C, Loundou A,
Moro-Sibilot D, Planchard D, Scherpereel A, Menu Y.
Inter- and intraobserver consistency in assessing eligibility for
bevacizumab (BVZ) in non-small-cell lung cancer (NSCLC) patients with
centrally located tumors.
Ann Oncol, 2010 Jan 11. [Epub ahead of print]
Holmberg K, Tonnel AB, Dreyfus I, Olsson P, Cougnard J, Mesbah K,
Devillier P.
Desloratadine relieves nasal congestion and improves quality-of-life in
persistent allergic rhinitis.
Allergy, 2009, 64:1663-1670.
Burgel PR, Paillasseur JL, Caillaud D, Tillie-Leblond I, Chanez P,
Escamilla R, Court-Fortune I, Perez T, Carré P, Roche N ; on behalf
of the Initiatives BPCO Scientific Committee.
Clinical COPD phenotypes : a novel approach using principal component
and cluster analyses.
Eur Respir J, 2010 Jan 14. [Epub ahead of print]
Hubert D, Le Roux E, Lavrut T, Wallaert B, Scheid P, Manach D,
Grenet D, Sermet-Gaudelus I, Ramel S, Cracowski C, Sardet A,
Wizla N, Deneuville E, Garraffo R.
Continuous versus intermittent infusions of ceftazidime for treating
exacerbation of cystic fibrosis.
Antimicrob Agents Chemother, 2009, 53:3650-3656.
Chang Y, de Nadai P, Azzaoui I, Morales O, Delhem N, Vorng H, Tomavo
S, Ait Yahia S, Zhang G, Wallaert B, Chenivesse C, Tsicopoulos A.
The chemokine CCL18 generates adaptive regulatory T cells from memory
CD4+ T cells of healthy but not allergic subjects.
FASEB J, 2010, Aug 11. [Epub ahead of print]
King TE Jr, Albera C, Bradford WZ, Costabel U, Hormel P,
Lancaster L, Noble PW, Sahn SA, Szwarcberg J, Thomeer M,
Valeyre D, du Bois RM ; Wallaert B for the INSPIRE Study Group.
Effect of interferon gamma-1b on survival in patients with idiopathic
pulmonary fibrosis (INSPIRE) : a multicentre, randomised, placebocontrolled trial.
Lancet, 2009, 374:222-228.
63
Contents
Deswarte G, Richardson M, Polge AS, Pouwels S, Ennezat PV,
Trochu JN, Wallaert B, Deklunder G, Le Tourneau T.
Longitudinal Right Ventricular Function as a Predictor of Functional
Capacity in Patients with Mitral Stenosis : An Exercise Echocardiographic
Study.
J Am Soc Echocardiogr, 2010 Apr 29. [Epub ahead of print].
Scherpereel A, Astoul P, Baas P, Berghmans T, Clayson H, de Vuyst P,
Dienemann H, Galateau-Salle F, Hennequin C, Hillerdal G,
Le Péchoux C, Mutti L, Pairon JC, Stahel R, van Houtte P,
van Meerbeeck J, Waller D, Weder W.
Guidelines of the European Respiratory Society and the European Society
of Thoracic Surgeons for management of Malignant Pleural
Mesothelioma.
Eur Respir J, 2009 Aug 28. [Epub ahead of print]
Just N, Bautin N, Danel-Brunaud V, Debroucker V, Matran R, Perez T.
The BORG dyspnoea score : a relevant clinical marker of inspiratory
muscle weakness in amyotrophic lateral sclerosis.
Eur Respir J, 2010, 35:353-360.
Scherpereel A, Berghmans T, Lafitte JJ, Colinet B, Richez M,
Bonduelle Y, Meert AP, Dhalluin X, Leclercq N, Paesmans M,
Willems L; J.P. Sculier for the European Lung Cancer Working Party
(ELCWP).
Valproate-doxorubicin : promising therapy for progressing mesothelioma. A phase II study.
Eur Respir J, 2010, Jun 7.[Epub ahead of print]
Leroy X, Aubert S, Zini L, Franquet H, Kervoaze G, Villers A,
Delehedde M, Copin MC, Lassalle P.
Vascular endocan (ESM-1) is markedly overexpressed in clear cell renal
cell carcinoma.
Histopathology, 2010, 56:180-187.
Tillie-Leblond I, Grenouillet F, Reboux G, Roussel S, Chouraki B,
Lorthois C, Dalphin JC, Wallaert B, Millon L.
Hypersensitivity pneumonitis and metalworking fluids contaminated by
Mycobacteria.
Eur Respir J, 2010, Aug 6.
Morawiec E, Hachulla-Lemaire AL, Chabrol J, Rémy-Jardin M,
Wallaert B.
Venoatrial compression by lymphadenopathy in sarcoidosis.
Eur Respir J, 2010, 35:1188-1191.
Mouillot G, Carmagnat M, Gérard L, Garnier JL, Fieschi C, Vince N,
Karlin L, Viallard JF, Jaussaud R, Boileau J, Donadieu J, Gardembas M,
Schleinitz N, Suarez F, Hachulla E, Delavigne K, Morisset M, Jacquot S,
Just N, Galicier L, Charron D, Debré P, Oksenhendler E, Rabian C
for the DEFI Study Group.
B-Cell and T-Cell Phenotypes in CVID Patients Correlate with the Clinical
Phenotype of the Disease.
J Clin Immunol, 2010, 30:746-755.
Torres D, Dieudonné A, Ryffel B, Vilain E, Si-Tahar M, Pichavant M,
Lassalle P, Trottein F, Gosset P.
Double-stranded RNA exacerbates pulmonary allergic reaction through
TLR3: implication of airway epithelium and dendritic cells.
J Immunol, 2010, 185:451-459.
Van Meerbeeck JP, Scherpereel A, Surmont VF, Baas P.
Malignant pleural mesothelioma : The standard of care and challenges
for future management.
Crit Rev Oncol Hematol, 2010, May 11. [Epub ahead of print].
Paesmans M, Berghmans T, Dusart M, Garcia C, Hossein-Foucher C,
Lafitte JJ, Mascaux C, Meert AP, Roelandts M, Scherpereel A,
Terrones Munoz V, Sculier JP; European Lung Cancer Working Party,
and on behalf of the IASLC Lung Cancer Staging Project.
Primary tumor standardized uptake value measured on
fluorodeoxyglucose positron emission tomography is of prognostic value
for survival in non-small cell lung cancer : update of a systematic review
and meta-analysis by the European Lung Cancer Working Party for the
International Association for the Study of Lung Cancer Staging Project.
J Thorac Oncol, 2010, 5:612-619.
PhD
Cécile CHENIVESSE
Directeur de thèse : Anne Tsicopoulos
Les chimiokines, actrices de la déviation immune.
11 décembre 2007
Pourcet B, Pineda-Torra I, Derudas B, Staels B, Glineur C.
SUMOylation of human peroxisome proliferator-activated receptor alpha
inhibits its trans-activity through the recruitment of the nuclear corepressor
NCoR.
J Biol Chem, 2010, 285(9):5983-5992
Stéphane SARRAZIN
Directeur de thèse : Maryse Delehedde
Caractérisation des interactions glycosaminoglycannes/protéines dans
le but de développer des molécules d’intérêt thérapeutique : exemples de
l’Endocan et de l'interféron gamma.
27 juin 2007
Raiko I, Sander I, Weber DG, Raulf-Heimsoth M, Gillissen A,
Kollmeier J, Scherpereel A, Brüning T, Johnen G.
Development of an enzyme-linked immunosorbent assay for the detection
of human calretinin in plasma and serum of mesothelioma patients.
BMC Cancer, 2010, 10:242.
Latiffa AMNIAI
Directeur de thèse : Catherine Duez
Régulation de la réaction allergique pulmonaire : inhibition par un BCG
recombinant dans un modèle murin et déficit de la réponse des cellules
Natural Killer de patients allergiques à la chimiokine CCL18.
17 décembre 2007
Reikvam H, Hatfield KJ, Lassalle P, Kittang AO, Ersvaer E, Bruserud Ø.
Targeting the angiopoietin (Ang)/Tie-2 pathway in the crosstalk between
acute myeloid leukaemia and endothelial cells : studies of Tie-2 blocking
antibodies, exogenous Ang-2 and inhibition of constitutive agonistic Ang1 release.
Expert Opin Investig Drugs, 2010, 19:169-183.
Nathalie de FREITAS CAIRES
Directeur de thèse : Philippe Lassalle
Etude de la dégradation d’endocan par les neutrophiles et implication
dans le sepsis.
19 décembre 2008
Sarrazin S, Lyon M, Deakin JA, Guerrini M, Lassalle P, Delehedde M,
Lortat-Jacob H.
Characterization and binding activity of the chonDroitin / dermatan
sulphate chain from Endocan, a soluble endothelial Proteoglycan.
Glycobiology, 2010, Jun 24. [Epub ahead of print].
64
Contents
Jules GILET
Directeur de thèse : Anne Tsicopoulos
Implication des chimiokines dans le développement des réactions
allergiques : apport des modèles murins.
18 décembre 2008
Benjamin LEGENDRE
Directeur de thèse : Philippe Lassalle
Etude comparative structurale et fonctionnelle de CCL18 recombinant
produit par la CHO et E.Coli.
10 mars 2010
Coline PLE
Directeur de thèse : Catherine Duez
Role des cellules NK dans l’asthme allergique.
2 avril 2010
Patents
Lassalle P, Grigoriu BD, Depontieu F, Marchandise G, Kervoaze G,
Janin A.
The use of a non-glycanated polypeptide for treating a cancer
Dépôt le 24 octobre 2007. Brevet BIO60226
de Freitas Caires N, Kervoaze G, Marchandise G, Lassalle P.
Marker peptides for determining the occurrence of an inflammatory state
in a subject
Dépôt le 8 octobre 2008. EP 08305655.6
65
Contents
Cancer
66
Contents
Genetic, Functional
and Structural
approaches
of cancer biology
CNRS UMR 8161
Yvan de LAUNOIT
Group Members Common Facilities :
Contact :
Secrétariat général
Boutin Philippe, Engineer Research CNRS
00 33 3 20 87 11 17
[email protected]
Secrétariat général de l'IBL et Ressources Humaines
Cluckers Francine, Technician CNRS
Joonekindt Cathy, Technician CNRS
Equipes Secrétariat Gestion de l'IBL
Lefebvre Frédéric, Assistant Engineer CNRS
Blanchet Régine, Technician CNRS
Bouchez Marie-Christine, Technician CNRS
Crampe Patricia, Technician CNRS
Romont Françoise, Technician CNRS
Devassine Nicole, Technician Inserm
Equipe Informatique de l'IBL
Oulmi Karl, Engineer CNRS
Bercker Geoffrey, Technician CNRS
Connart Guilllaume, Student CNRS
Equipe Technique et Logistique de l'IBL
Caplier Hugues, Assistant Engineer CNRS
Messemanne Marc, Technician CNRS
Vitoux Jean-Yves, Technician CNRS
Leman Jerome, Technician CNRS
Equipe Laverie/Milieux centralisés de l'IBL
Pattin Michèle, Technician Inserm
Deffrasnes Daniel, Agent Technique Inserm
Hamy Sylvie, Agent Techique IPL
Lecocq Nadine, Agent Technique IPL
Fleurbaix Marie Andrée, Agent Technique CNRS
Chargée de la Mise en OEuvre des règles d'hygiène et de sécurité pour l'UMR8161 et le
bâtiment IBL
Roland Isabelle, Technician IPL
Key Words :
Cancer. c-MET. Apoptosis. Senescence. NF-kappaB. Metastasis.
PEA3. HIC1. Biostructure. Chemistry of Biomolecules.
Angiogenesis. Ets.
67
Contents
Cancer
Teams
* Structural and Functional Approaches of Pathogenesis
Vincent VILLERET
Virus, Cancer and Transcription
Yvan de LAUNOIT
* Identification and Characterisation of New Genetic Events Involved in
an early Step of Tumor Development : the Senescence Escape
David BERNARD
Functional Analyses of the tumor suppressor HIC1
Dominique LEPRINCE
VE-statin/egf17 and vascular development
Fabrice SONCIN
Cancer Biology and Chemistry
Oleg MELNYK
Initiation of epithelial cancers
Corinne ABBADIE
* Parvovirus : Oncosuppression and Gene Therapy
Dominique STEHELIN
Signalling, Apoptosis and Cancer
David TULASNE
* Pharmacochemistry and Proteomics
Patricia MELNYK
* Team are no longer part of UMR 8161 since Juanary 1st, 2010
68
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Cancer
2013 period to reinforce the links between the three cancer
research centers to finally propose a joint program in 2013.
Research Report
In the present project, a special attention will be given to
reinforce the interdisciplinary programs between the chemists
and biologists, as well as between the “structuralists” and
biologists of the UMR 8161. This is for example the case of the
strong collaboration between the biologists studying the
HGF/SF receptor, MET, and the chemists specialized in sugar
potential inhibitor (deregulated MET signaling is involved in
tumor progression). This collaboration has resulted in the
generation of a multidisciplinary research team: team 1. This
research has been supported by the CNO. The three groups
will contribute in a complementary program on the discovery
of new extracellular MET inhibitors : O. Melnyk (chemical
synthesis of polysaccharides and high throughput screening),
V. Fafeur (biological activity of the compounds : kinase and
heparanase assays, proliferation, survival, invasion,
morphogenesis) and E. Adriaenssens (pre-clinical in vivo
models). In short, after initial screenings of polysaccharides by
chemists and biologists, it will be necessary to determine the
mode of action of the selected inhibitory polysaccharides. The
identification of compounds able to inhibit HGF/MET
signaling and/or heparanase activity should lead to highly
promising anticancer therapies. The team of D. Tulasne (team
6) will study some fundamental aspects of the HGF/MET
pathway. More particularly, they will study the initial events
regulating the MET receptor ; i.e. the proteolytic processing
during development and tumorigenesis
From 2006-2009, parts of the teams within the IBL building
have been merged in one research unit: UMR 8161, “Institut
de Biologie de Lille”. The research conducted in the unit has
been mainly focused on cancer and microbiology in close
interaction with chemists and biostructuralists. Since the
discovery of the first oncogenes in Lille, a strong activity of
research in the field of cancer has been held in UMR 8161; i.e.
the identification of molecular mechanisms (from membrane
receptors to transcription factors) by which a normal cell
becomes tumoral, and finally metastatic. Several of these
research teams are currently involved in the North-West
“Canceropole”. The second research topic concerned structural
studies of membrane structure and transport, focused so far
on bacterial systems. The third research topic concerned the
major steps of hepatitis C virus life cycle and how this virus
interacts with hepatocytes. Finally, a fourth topic was also
developed in the field of cellular biology of the early steps of
bacterial infection; this allowed the development of new
confocal microscopic tools applicable to cancer research.
These last two microbiology teams will join the future “Centre
of Infection and Immunity of Lille” in 2010. In parallel, the
Laboratory of Genetic Toxicology of the “Institut Pasteur de
Lille” was part of the research group EA2690 “Occupational
and environmental toxicants and carcinogens: new markers
and health effects” whose aim was to develop and optimize
models of genotoxicity in order to better understand
mechanisms of action, and to apply the developed models to
the study of environmental hazard/risk assessment. This group
will join the future team “Impact of environmental chemicals
on human health” in 2010.
In the next 4 years we will continue our efforts to support
interdisciplinary projects specifically, for example by upgrading the technological platforms. A particular effort will be
made on the support of team 2. In fact, basic research on the
epigenetic of cancer will be one of the main lines of this
project. More particularly, the major goals of the team of D.
Leprince will be to pursue the study on the precise mechanism
brought by the product of a tumor suppressor gene on the
transcriptional repression of its target genes. More specifically,
they will decipher the epigenetic modifications of HIC1-target.
This will be correlated with HIC1-induced tumorigenesis in
animal models, as well as in human tumors. We will do our
best to recruit young scientists in order to develop new
research programs with D. Leprince. Recent data on
epigenetic changes in cancer also concern the link between
DNA methylation and histone modifications on the one hand
and expression of non-coding RNAs on the other hand. E.
Adriaenssens (team 1) will focus part of his work on the role
of the 91H antisense non-coding-RNA (H19/IGF2 locus) on
tumorigenesis.
The first steps of tumorigenesis will also be depicted through
the angle of the escape from senescence or programmed cell
death. The research teams will pursue their work on the
characterization of the molecular mechanisms by which a
normal cell becomes pre-tumoral by escaping senescence.
C. Abbadie (team 4) will address this topic by searching for
genomic alterations in senescent cells induced by their
endogenous oxidative stress or by the microenvironment and
which could contribute to senescence escape. In parallel, they
will also investigate whether the emergence of transformed
cells necessitates escaping autophagic cell death. Finally, their
interest will be focused on post-senescent emergent cells that
can be cancer stem cells.
Perspectives and
Development
Scientific project
For the next 4-year period, the UMR 8161 interest will be
focused on cancer research. For this purpose, the unit is
composed of 6 independent and interactive teams. Most of
these teams are involved in INCa and ANR programs, as well
as in the North-West Canceropole (CNO) program, Y. de Launoit
being board member of the CNO.
In order to reinforce the cancer research visibility in the Nord
– Pas de Calais region, a special effort has been made since
2004 to favor joint programs and collaborations between the
regional teams performing different aspects of cancer
research; i.e. the J.P. Aubert Research Center at the University
of Lille 2 on the CHR campus, different units at the University
of Lille 1 (USTL) and the UMR 8161 located on the Pasteur
Institute campus. This partnership permits, for example, to
propose a research program on the molecular basis of colon
cancer metastasis. Moreover, the support on cancer research
by the Nord – Pas de Calais region is shown by multiple PhD
grants and special effort on research programs, as well as
expensive equipments. We will take advantage of the 2010-
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Cancer
The knowledge of the first steps of tumorigenesis is crucial to
fight cancer more efficiently. Furthermore, tumoral neoangiogenesis is also a key phenomenon prior to metastasis
development. The team of F. Soncin (team 3) will pursue their
work on the role of VE-statin in tumor angiogenesis.
Supported by a “labellisation” of the “Ligue Nationale Contre
le Cancer”, this research team will specifically develop new
mouse pre-clinical models.
The interaction with the clinics will be reinforced, more
particularly thanks to the teams studying the immune
response in EBV- and HCV-related tumors. Hence, team 5 (Y.
de Launoit) will dissect the immunosuppression mechanism
in HCV-induced hepatocarcinoma. Moreover, this team will
pursue the use of the H-1 Parvovirus on hepatocarcinoma as
a specific oncolytic treatment.
Team 6 of D. Tulasne will receive the help of the group of A.
Chotteau-Lelièvre to dissect the links between the HGF/SF
pathway and the PEA3 transcriptional program in the
metastatic process.
The significant mobilization of our forces around bacterial
systems has not prevented us from exploring new directions
recently. Thus, lately, we launched a new research program
which contrasted with the general "historical" themes of the
laboratory. Our motivation was the desire to reinforce our
activities toward fields corresponding to patent social
requests. Moreover, the Transcription-Development-Cancer
team (team 2), headed by Y. de Launoit, was looking for
expertise in the field of structural biology in order to progress
in understanding transcriptional regulatory processes of
factors involved in cancer metastasis. This allowed the setting
up of a timely collaboration between the two teams in the
field of cancer, as the structural characterization of eukaryotic
transcription factors requires not only approaches by
diffraction techniques, but also the solution approaches
recently implemented in our laboratory. The project is
boosting, with the recruitment in 2008 of a CNRS research
associate, A. Verger, as well as grants from different origins.
Type V secretion : the Two-Partner Secretion (TPS) pathway
The targeting of proteins to their dedicated subcellular
compartments is essential for cell function and organelle
biogenesis. The translocation of proteins across or insertion
into membranes is mediated by protein machineries some of
which have been conserved throughout evolution, such as the
transporters of the Omp85/TpsB superfamily. TpsB
transporters are components of TPS systems, the most widely
distributed secretion mechanism known, which is devoted to
the secretion of large, mostly -helical proteins serving
generally as virulence factors in Gram-negative bacteria and
collectively called “TpsA” proteins. The superfamily also
includes the Toc75, Sam50/Tob55 and Omp85/YaeT
homologs, which are the cores of large hetero-oligomeric
complexes involved in protein transport across, and in
insertion of -barrel proteins into the outer membranes of
chloroplasts, mitochondria and Gram-negative bacteria,
respectively. Omp85/TpsB transporters have all been
predicted to be composed of a conserved C-terminal
transmembrane -barrel and a soluble N-terminal region
harboring 1 to 5 putative polypeptide-transport associated
(POTRA) domains, hypothesized to mediate protein-protein
interactions. They also harbor conserved C-proximal signature
motifs of unknown function in their pore-forming regions.
A hallmark of the TPS pathway is also the presence in the TpsA
proteins of a conserved N-proximal module called the TPS
domain, which is essential for secretion. The TPS domain is
thus hypothesized to bear secretion determinants mediating
specific interactions of TpsA proteins with their TpsB
transporters at the periplasmic side of the outer membrane.
These recognition events are thought to trigger the
translocation of the TpsA proteins, starting from their Nterminus, across that membrane.
In spite of their implication in critical physiological processes
such as membrane biogenesis and secretion of virulence
proteins, the molecular mechanisms of protein translocation
or integration by those transporters remain poorly understood
and in particular, no X-ray structure was available for any of the
partners of such secretion systems when we initiated our
studies. To address these issues, we have studied, in
collaboration with microbiologists, the Tps prototype system
FhaC/FHA that mediates the translocation to the bacterial
surface of filamentous hemagglutinin (FHA), the major adhesin
of the whooping cough agent Bordetella pertussis.
The composition of the UMR8161 for the next four years
is thus :
1. Cancer biology and chemistry (Oleg Melnyk)
2. Functional studies of the tumor suppressor gene HIC1
(Dominique Leprince)
3. VE-statin/egfl7 functions during physiological and tumoral
vascular development (Fabrice Soncin)
4. Initiation of epithelial cancers (Corinne Abbadie)
5. Virus-cancer and transcription (Yvan de Launoit)
6. Signalling, apoptosis and cancer (David Tulasne)
Structural and Functional
Approaches of
Pathogenesis
Vincent VILLERET
DR2 - CNRS
Since its creation in 2002 as an ATIP of the CNRS, the main focus
of our lab has been to gain insights into the structural and
functional aspects of the virulence of bacterial pathogens that
are closely related to membrane systems. Since then we have
used X-ray diffraction as a main technique, to study stable and
structured proteins or domains. Many proteins that are
emerging in connection with bacterial virulence appear to
possess structural heterogeneities resulting from a multidomain topology and/or the presence of intrinsically
unstructured regions. Thus their structural characterization
cannot be achieved solely by diffraction techniques but
requires the use of other methods. We have recently acquired
new skills, allowing “multifaceted” approaches of biological
problems, including molecular biology, biochemistry,
bioinformatics and the use of biophysical tools such as light
scattering, circular dichroïsm and SAXS, a powerful technique
allowing the determination of molecular shapes at low
resolution (around 10-15 Å) and well suited for the study of
partially folded proteins.
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Cancer
We have determined i) the crystal structure of the TPS domain
of FHA and ii) the crystal structure of the transmembrane
transporter FhaC that mediates specifically the translocation
to the bacterial surface of FHA. FHA transits through the
periplasm in an extended conformation before its transport
across the outer membrane by FhaC and as such, must be
protected from degradation. Recently, the periplasmic
chaperone Par27, the prototype of a new group of parvulins,
was shown to bind to FHA fragments. Par27 also displays
affinity for other proteins rich in amphipathic structure such
as outer membrane porins, and therefore, it has been
proposed to serve as a general periplasmic chaperone in
B. pertussis. We have also determined the structure of Par27,
using a combination of X-ray crystallography, SAXS and
modelling analyses.
All these data have been complemented by site-directed
mutagenesis studies, biochemical assays and topological
studies. These results have already been published (Clantin et
al., PNAS USA 101: 6149; 2004 - Hodak et al., Mol Microbiol 61:
368; 2006 - Méli et al., J Biol Chem 281: 158; 2006 - Clantin et al.,
Science, 317: 957; 2007 - Wohlkönig et al., Acta Cryst Struct Biol
Com In press ; 2008 - Clantin et al., J Struct Biol Submitted). Our
long term objective is to unravel at the structural and functional
level the secretion process mediated by TPS systems, and
further extend our research to the Omp85/TpsB superfamily.
The BvgA-BvgS TCS is a signal transduction device responding
to changing growth conditions. We have initiated the study of
the periplasmic part of the inner membrane regulator BvgS,
which is constituted by two covalently linked "PBP-like"
domains. We have determined the high resolution structure
of one of this PBP domain. We have also crystallized the fulllength periplasmic part, but obtained twinned crystals
unsuitable for structure determination. We have now obtained
the global structure of the full-length periplasmic part by a
combination of X-rays, SAXS and modelling techniques (In
preparation).
B) heparin-binding haemagglutinin (HBHA), the major adhesin
in Mycobacterium tuberculosis virulence.
M. tuberculosis, the worldwide leading causative agent of
death owing to a single etiologic agent, adheres to epithelial
cells via the HBHA, a 199-residue protein that recognizes
heparan sulphate proteoglycans (HSPG). HBHA has been
shown to be involved (i) in the mycobacterial interaction with
epithelial cells, but not with professional phagocytes and (ii)
in the extrapulmonary dissemination of the bacilli by a
mechanism that still remains to be explained. The HBHAmediated adherence seems to rely at least in the interaction
of its C-terminal lysine-rich domain with HSPG receptors
present on the surface of its target cells. But in spite of its
crucial implication in M. tuberculosis virulence, this adhesin has
been only poorly characterized. We have tried to determine
its X-ray structure, having succeeded in producing large
amounts of highly pure and stable protein. All our
crystallization trials proved unsuccessful. By using various
approaches such as circular dichroïsm, cross-linking
experiments, light scattering and SAXS, we were able to show
that the protein contains large unstructured regions (probably
preventing crystallization). Using these techniques, we have
determined a low resolution molecular envelope of HBHA. Our
studies have also shown the importance of the disordered
regions in the recognition process of the adhesin with its
various ligands. We have also shown that HBHA shrinks during
its interaction with a sulfated disaccharide, which mimics
sulfated chondroitines present at the surface of epithelial cells.
This shrinking could be involved in the phagocytic process of
the bacilli. These results allow to progress in understanding
the interactions of HBHA with its ligands, despite the
unstructured nature of the protein. These results have still to
be published. Preliminary data have been reported (Dupres et
al., Nature Methods 2: 515; 2005 - Verbelen et al., submitted).
Regulation and effectors of virulence
A) Two Component Systems
The import of nutriments and solutes is required for bacterial
survival and adaptation to environmental conditions. They may
also represent various modulators of virulence, via for example
the two-component signal transduction pathway, a common
mechanism used by bacteria to sense changes in their
environment and controlling the synthesis of virulence
effectors. In Gram-negative bacteria, these molecules must
transit via the periplasm and, in ABC, Trap or TTT transporters,
their import involves various periplasmic binding proteins
(PBPs) which bind specific ligands and deliver them to their
respective inner membrane partners. Many such proteins have
been identified in Bordetella pertussis, and some of them are
among the most abundant periplasmic proteins, suggesting
important but yet poorly characterized function. Hence these
last years our team has structurally and functionally
characterized, in collaboration with microbiologists, so-called
"Bug" proteins from B. pertussis, which form a large family of
periplasmic solute-binding receptors. These structures reveal
high conservation of the Bug architecture, despite limited
sequence identity. They display a specific ligand-binding motif
highly conserved and designed to accommodate carboxylated
solutes. The vast expansion of the Bug family in several
bacterial genera is likely to be explained by the possible
diversity of ligands. We also characterized PBPs potentially
involved in the regulation of virulence in B. pertussis : Bug27,
Dctp6 and Dctp7. These studies allowed to propose
mechanisms of ligand - protein interactions, and also paved
the way to the study of the Two-Component System BvgABvgS, the master regulator of virulence controlling virtually all
known virulence traits of B. pertussis, which also involves PBPlike domains. These studies on PBPs have been published
(Huvent et al., Acta Cryst D Biol Crystal 62: 1375; 2006 - Huvent
et al., J Mol Biol 356: 1014; 2006 - Rucktooa et al., Acta Cryst F
Biol Crystal 62: 970; 2006 - Rucktooa et al., J Mol Biol 370: 93;
2007 - Herrou et al., J Mol Biol 373: 954; 2007).
C) Type Three Secretion Systems (T3SS)
Historically we had a long interest in the study of virulence
effectors translocated in host cells by T3SS but the structural
analysis of a major effector of Salmonella enterica really started
with our moving into the IBL in 2002. The invasion of epithelial
cells by S. enterica is mediated by bacterial "effector" proteins
that are delivered into the host cell by T3SS. Although
primarily known for their roles in actin rearrangements and
membrane ruffling, translocated effectors also affect host cell
processes that are not directly associated with invasion.
SopB/SigD, an effector with phosphoinositide phosphatase
activity, has anti-apoptotic activity in Salmonella-infected
epithelial cells. The crystallographic study of the phosphatase
domain of SopB could not only provide the first structural
characterization of an inositol 4-phosphatase, it could also
open the way to a detailed structure-function study of such
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Cancer
phosphatase. These results would provide new insights into
the mechanisms of apoptosis and highlight how bacterial
effectors can intercept signaling pathways to manipulate host
responses. Type III effectors require the presence in the
bacterial cytosol of specific TTS chaperones. During the 20022004 period, we co-expressed SopB with its specific
chaperone SigE and purified the complex but no crystals
could be obtained probably due to conformational
heterogeneities in the complex. We thus mapped the different
structural domains of the complex by limited proteolysis. This
enabled us to define the limits for the chaperone-binding
domain as well as for the C-terminal catalytic domain of SopB
which have been subsequently produced in E. coli and
purified. The structure of SigE had been previously solved by
others and the constructs of several domains of SopB were
also available but no crystals could ever be obtained.
We have now turned towards a structural approach in solution
for the whole complex as well as for the several constructions
designed while mapping the domains. In that way we recently
collected SAXS data at the ESRF in Grenoble that are currently
processed (PhD work of P. Lebrun) in order to describe the
interaction between SopB and its chaperone and the
quaternary structure on the complex. A manuscript describing
the first step of this work is in preparation.
For biophysical studies, the first bottleneck to overcome is to
produce enough material. We thus screened a large number
of constructs for the expression of Erm in bacteria, and we
recently succeeded in overexpressing Erm1-122 (see below),
which encompasses the complete transactivation domain
including the first SUMOylation site. In collaboration with
team 2, we have initiated its structural characterization and
the effect of its SUMO modification on lysine 89. To achieve
this, we also needed to develop protocols allowing the
sumoylation of overexpressed proteins in bacteria.
Numerous constructs including up to residue 370 of Erm (just
before the DNA binding domain) were tested for expression
in bacteria, and we were able to produce and purify in high
yields 6 His-tagged Erm1-122. To sumoylate this fragment on
lysine 89, we co-expressed it in bacteria with the components
of the sumoylation pathway (the E1 activating enzyme
Aos1/Uba2, the E2 conjugating enzyme Ubc9 and SUMO-1).
To do so, we have used the bacterial vector pT-E1E2S1
developed by Hisato Saitoh, who solved the structure of the
sumoylated thymine DNA glycosylase (Baba et al., Nature 435:
979; 2005). We have observed significant level of ERM
modified by SUMO, indicating that the E. coli modification
system is effective. After 3 steps of purification (Ni-NTA [Ni2+
affinity], MonoQ [anion-exchange] and gel filtration [size
exclusion] chromatographies) and concentration, we have
obtained few mgs of pure Erm (1-122) that could be
concentrated up to 15mg/ml and also pure Erm (1-122)
modified by SUMO, concentrated up to 8mg/ml. After scaleup, we expect that purified Erm (1-122) and Erm-SUMO will be
suitable soon in amounts for structural studies combining CD,
light scattering, FTIR, and Small-angle X-ray scattering (SAXS)
analysis. Crystal screening will also be attempted, although
not much can be expected if the unstructured character of this
domain is confirmed by our biophysical studies.
Structural / functional relationship of the PEA3 group
transcription factors
As described below in the report of team 2, the PEA3 group of
human transcription factors is composed of three members:
Pea3, Er81 and Erm, which are key players in the transcriptional
regulation of enzymes involved in breast metastasis (de Launoit
et al., BBA 1766: 79; 2006). They all possess an ETS DNA binding
domain, two conserved transactivating domains (TADs) lying
respectively at the N and C termini and a negative regulatory
domain (NRD) flanking the acidic N-terminal transactivation
domain (TADn). If the structure of DNA binding domains has
been thoroughly investigated and structural classes clearly
defined, in sharp contrast, the structural constraints put on
transactivation regions is poorly understood. In PEA3 members,
as in many eukaryotic transcription factors, these regions are
predicted to contain large disordered segments (Mauen et al.,
BBA 1760: 1192; 2006) which are hypothesized to be structured
upon the recruitment of partners and/or post-translational
modifications. Regarding the PEA3 factors, team 2 has initiated
functional studies aiming at identifying transcriptional partners
of Erm and also at characterizing post-translational
modifications regulating its function. This team has shown
recently that SUMO modifications of the NRD play crucial roles
in the regulation of Erm. Erm sumoylation causes inhibition of
TADn transcription-enhancing activity. TADn extends to residue
122 and overlaps from residue 73 with the NRD domain, which
extends up to residue 298. The NRD domain contains three
SUMO sites, one of which being in the overlapping segment
with TADn on lysine 89. The current view is that Erm does not
contain a negative regulatory region per se but multiple short
inhibitory modules corresponding to sumoylation motifs. TAD1122 is thus a region of high interest, as it contains the complete
acidic transactivation domain as well as the first sumoylation
motif, able to influence TAD activity. How SUMO works to alter
TAD structure and function is unknown. SUMO may interfere
with the recruitment of co-factors by decreasing the interaction
with transcriptional elements required for transcriptional
activity.
Since 2004, as a regional X-ray crystallography platform, our
research team has also collaborated on different research
programs linked to structural biology and drug design.
- Side effects associated with tuberculosis therapy brings with
it dangers of non-compliance and subsequent drug resistance. Increasing the therapeutic index of antituberculosis
drugs should thus improve treatment effectiveness. Several
antituberculosis compounds require in situ metabolic
activation to become inhibitory. Various thiocarbamidecontaining drugs, including ethionamide (ETH), a well known
second-line drug, are activated by EthA, the production of
which is controlled by the repressor EthR.
In 2002, we initiated a fundamental study of this repressor, in
order to understand how it could bind cooperatively to its
target DNA and form an unusual octamer. In 2004, we reported
the crystal structure of EthR without bound DNA. The structure
was found to be fortuitously liganded, resulting in a
conformational state of EthR incompatible with repressor
function. Such a conformation would lead in vivo in ethA
derepression and consequently to an increased sensitivity to
ethionamide and other thioamides. This led us to propose a
strategy, based on the crystal structure, to increase the
sensitivity of M. tuberculosis to ETH. This strategy would broaden
its therapeutic window and render it effective at lower dosages,
minimizing side effects and allowing its use as a first line drug
(Frénois et al., Mol Cell 16: 301; 2004 - Frénois et al., Tuberculosis
86: 110; 2006). In collaboration with pharmacochemists (B.
Déprez, Lille2) and microbiologists (A. Baulard, IPL), we took part
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Cancer
Wohlkönig A, Sénéchal M, Dewitte F, Backers K, Emeux C, Villeret V.
Expression and purification in high yield of a functionally active
recombinant human Type I inositol(1,4,5)P3 5-phosphatase.
Protein Expr Purif, 2007, 55:69-74.
in a drug design program aiming at the discovery of synthetic
EthR inhibitors boosting antituberculosis activity of
ethionamide. Our goal was to identify drug-like inhibitors of
EthR to boost the bio-activation of ethionamide. Compounds
designed and screened for their capacity to inhibit EthR-DNA
interaction were co-crystallized with EthR. 3D-structures were
exploited for the synthesis of improved analogs that boosted
more than 10 fold the ethionamide potency in culture. In
Mycobacterium-infected mice, a substantially reduced dose of
ethionamide associated with compound BDM31343 lessened
the mycobacterial load as efficiently as the conventional
treatment. This provides proof-of-concept that inhibiting EthR
improves the therapeutic indexes of thiocarbamide-derivatives,
permitting to reconsider their use as first line drugs. The
publication has been submitted to Nature Medecine. Patents
covering these findings have been issued.
- In collaboration with Dr. Wintjens at the ULB, we have
determined and analyzed the structures of a glutaminyl
cyclase and a chitinase isolated from papaya latex (Azarkan
et al., Acta Crystal F Struct Biol Cryst Com 61: 59; 2005 Wintjens et al., J Mol Biol 357: 457; 2006 - Huet et al., Acta
Crystal F Struct Biol Cryst Com 64: 371; 2008 - Huet et al.,
Biochemistry In press; 2008).
- In collaboration with J.P. Bohin at the USTL, we have
determined the structure of a glycosyltransferase involved in
osmoregulated periplasmic glucans in the cell wall of gramnegative bacteria (Hanoulle et al., J Mol Biol 342: 195; 2004).
- We finalized a functional study on inositol phosphatases
that had been conducted in collaboration with C. Erneux at
the ULB. Two papers were published on this topic
(Vandeput et al., Cell Signal 18: 2193; 2006 - Wohlkönig et
al., Protein Expr Purif 55: 69; 2007).
- Recently, we contributed structural analyses, in
collaboration with J.C. Sirard, from IPL, on a bacterial
flagellin (Nempont et al., J Immunol 181: 2036; 2008) and
with V. Fafeur (team 6) on the MET tyrosine kinase receptor
(manuscript in preparation).
2008
Huet J, Azarkan M, Looze Y, VilleretV, Wintjens R.
Crystallization and preliminary X-ray analysis of a family 19 glycosyl
hydrolase from Carica papaya latex.
Huet J, Rucktooa P, Clantin B, Azarkan M, Looze Y, Villeret V,
Wintjens R.
X-ray Structure of Papaya Chitinase Reveals the Substrate Binding Mode
of Glycosyl Hydrolase Family 19 Chitinases.
Biochemistry, 2008, 47:8283-8291.
Deletion of flagellin's hypervariable region abrogates antibody-mediated
neutralization and systemic activation of TLR5-dependent immunity.
J Immunol, 2008, 181:2036-2043.
Wohlkönig A, Hodak H, Clantin B, Sénéchal M, Bompard C, JacobDubuisson F, Villeret V.
Crystallization and preliminary X-ray diffraction analysis of the PeptidylProline Isomerase Par27 of Bordetella pertussis.
2009
Clantin B, Leyrat C, Wohlkönig A, Hodak H, Ribeiro ED Jr, Martinez N,
Baud C, Smet-Nocca C, Villeret V, Jacob-Dubuisson F, Jamin M.
Structure and plasticity of the peptidyl-prolyl isomerase Par27 of
Bordetella pertussis revealed by X-ray diffraction and small-angle X-ray
scattering.
J Struct Biol, 2009, 169:253-265.
Deheuninck J, Goormachtigh G, Foveau B, Ji Z, Leroy C, Ancot F,
Villeret V, Tulasne D, Fafeur V. 2009
Phosphorylation of the MET receptor on juxtamenbrane tyrosine residue
1001 inhibit its caspase-dependent cleavage.
Cell Signal, 2009, 21:1455-1463.
Publications
First structural insignts into the TpsB/Omp85 superfamily.
Biol Chem, 2009, 390:675-684. Review.
2007
Willand N, Dirié B, Carette X, Bifani P, Singhal A, Desroses M, Leroux F,
Willery E, Mathys V, Déprez-Poulain R, Delcroix G, Frénois F,
Aumercier M, Locht C, Villeret V, Déprez B, Baulard AR. 2009
Nat Med, 2009,15:537-544.
Clantin B, Delattre AS, Rucktooa P, Saint N, Albano C, Locht C, JacobDubuisson F, Villeret V.
Structure of the membrane protein FhaC : a member of the Omp85/TpsB
Science, 2007, 317:957-961.
PhD
Herrou J, Bompard C, Antoine R, Leroy A, Rucktooa P, Hot D, Huvent
I, Locht C, Villeret V, Jacob-Dubuisson F.
Structure-based mechanism of ligand binding for periplasmic solutebinding protein of the Bug family.
J Mol Biol, 2007, 373:954-964.
Prakash RUCKTOOA : PhD « Biologie-Santé »
Directeur : V. Villeret
Septembre 2007
J Mol Biol, 2007, 370:93-106.
Verbelen C, raze D, Dewitte F, Locht C, Dufrêne YF.
interactions.
J Bacteriol, 2007, 24:8801-8806.
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Cancer
Int J Cancer 70 : 590; 1997 - Chotteau Lelièvre et al., Clin Cancer
Res 10 : 7297; 2004). PEA3 group overexpression is correlated
to an increase of the expression of the extracellular matrix
degrading enzymes, the MMP ; thus permitting tissue
rearrangement, a common phenomenon in cancer metastasis.
In reporter assay experiments, the PEA3 group members
enhance transcription from different MMP promoters, such as
human stromelysin-1, the type I and IV collagenases and
matrilysin promoters. In vivo, the ectopic expression of Er81 in
the normal mouse mammary gland induces up-regulation of
the urokinase plasminogen activator gene. This ability to
enhance such promoters suggests that they could play a role
in tumorigenesis. We have recently reviewed the crucial
molecular roles of these factors in enhancing tumor growth,
invasion and metastasis (for review, see de Launoit et al., BBA
1766: 79; 2006). Moreover, our very recent data indicate that
when these factors are down-regulated in metastatic
mammary cancer cells, the capacities of the latter to
proliferate, migrate and invade are drastically decreased. In
fact, according to the expression of PEA3 transcription factors
in branching mammary glands, we know that murine
mammary ‘normal’ cells overexpressing Erm and Pea3 factors
have the capacity to spontaneously form an arborescence by
epithelial branching, which mimics physiological HGF/SF
stimulated morphogenesis (Chotteau-Lelievre et al., Dev Biol
259 : 241; 2003). On the other hand, murine mammary cancer
cells in which the expression of these factors is inhibited by
RNA interference display reduced proliferation, migration
and invasion. These cells injected subcutaneously to
immunodeficient mice form tumors whose size is significantly
reduced compared to those induced by parental cells (Firlej et
al., J Cell Science, In press). Thus we recently demonstrated the
involvement of PEA3 factors in the process leading to
mammary morphogenesis and tumorigenesis. To further
explore the molecular mechanism regulated by Pea3/Erm to
induce physiological epithelial morphogenesis and
tumorigenesis, we performed transcriptome microarray
analysis. An initial microarray study (388 genes) allowed the
identification of the pro-apoptotic bax gene as an Erm target
gene (Firlej et al., J Biol Chem 280 : 887; 2005). We now use a
new generation of transcriptomic analysis from Applied
Biosystems (28.218 validated mouse genes) supplemented by
a biostatistical analysis, developed by the team of A. Benecke
(IRI) with whom we collaborate on this project. By silencing
the PEA3 transcription factors in cancer mammary cell line, we
established a list of target genes regulated by these factors. A
significant part of them correspond to genes known to be
involved in the process of migration, invasion and cell
proliferation reinforcing the notion that these factors are key
regulators of epithelial invasion. We are currently extending
these experiences to the model of normal epithelial cells. The
challenge of this work is to compare genetic programs of
normal and transformed cells in order to characterize the
aberrant transcriptional regulations leading to tumorigenesis
(PhD work of V. Firlej [2003-2006] and F. Ladam [2007]).
Virus, Cancer
and Transcription
Yvan de LAUNOIT
DR1 - CNRS
Group Members :
Immunotherapy of viro-induced cancers (Nadira Delhem
and Yvan de launoit)
Delhem Nadira, MCU Univ Lille 1
Pancré Véronique, CR Inserm
Boleslawki Emmanuel, PR CHRU Lille
Caillet-Fauquet Perrine, CR
Martin Nathalie, Postdoctoral fellow CNRS
Morales Olivier, Engineer CNRS
Senechal Magalie, Technician CNRS
Richard Audrey, PhD Student MENRT
Miroux Céline, PhD Student CNRS/ANRS
Mrizak Dhafer, PhD Student BDI-PED/CNRS
Vausselin Thibaut, M2R Student
Ets proteins, transcriptional regulation and associated diseases
(Martine Duterque)
Duterque Martine, CR1 CNRS
Holder-Espinasse Muriel, MCU CHRU Lille (Mission permanente)
Flajollet Sébastien, Postdoctoral fellow CNRS/ANR
Flourens Anne-Claire, Technician Inserm
Tomavo Nathalie, Technician IPL
Tian Tian, PhD Student IPL-Région
Technical Platform (Yvan de Launoit)
Dumont Patrick, Engineer CNRS
Structure/Function
Relationship of the PEA3
Group Transcription Factors
Yvan de LAUNOIT
DR1 - CNRS
Role in the regulation of mammary metastasis
The study of the transcriptional regulation of enzymes
involved in breast metastasis is currently a key issue in breast
cancer research. The Ets proteins constitute a family of
eukaryotic transcription factors involved in development and
tumorigenesis. These proteins share a common ETS domain
responsible for DNA binding. Within this family, the PEA3
group is composed of three members, Pea3, Er81 and Erm,
which possess the ETS domain and two conserved
transactivating domains. In contrast, a relatively conserved
central domain (CIDD) of these proteins is responsible for
inhibition of transcriptional activity and DNA-binding. These
three factors are involved in mammary cancer and more
particularly in the regulation of the metastatic process. Hence,
in transgenic mice carrying several oncogenes, the PEA3
group members are overexpressed in metastatic mammary
tumors (for review, see de Launoit et al., BBA 1766: 79; 2006).
In human, these factors are also overexpressed in metastatic
primary cancers as well as in metastatic cell lines (Baert et al.,
Post-translational modifications of the PEA3 group
transcription factors
As for the other Ets proteins, these transcription factors
regulate the transcription of their target genes by subtle
molecular mechanisms. They, in fact, interact with general
transcriptional partners such as basal transcription complex
proteins and transcriptional co-activators (CBP/p300 for
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Cancer
example). They also synergize with transcription factors, such
as for example nuclear receptors or bHLH family proteins. The
PEA3 group members also undergo post-translational
modification that regulates their transactivation capacity. The
most common modification found in this group of proteins is
phosphorylation, as they are targets of the MAPK pathway
including Ras, Raf-1, MEK, ERK-1, and ERK-2. The
phosphorylation of specific serine and threonine residues
generally increases the transactivation capacity of these
transcription factors. Moreover, Erm and Er81 are also
phosphorylated through the PKA-mediated pathway (for
review, see de Launoit et al., BBA 1766: 79; 2006 – de Launoit
et al., Bull Mem Acad R Med Belg 162: 299; 2007)
research interest on the RING protein COP1 (constitutive
photomorphogenic 1). We found that the consensus
interacting domain of COP1 is found in the PEA3 group
members. COP1 is known to mediate ubiquitinylation of
target proteins, such as p53 and c-Jun (Yi & Deng, Trends Cell
Biol, 15: 618; 2005). For the latter, COP1 functions as an
adaptator protein which recruits the transcription factor to the
E3 complex, which contains other molecules such as DDB1,
culin4A, Roc1 and DET1 with which COP1 directly interacts.
We then proved biochemically that COP1 interacts with the
PEA3 group members; COP1 is thus requested to regulate the
transcriptional activity of the PEA3 group members (Baert et
al., submitted).
Finally, these transcription factors also cooperate with several
co-partners to regulate the transcription of their target genes.
For this purpose, we engaged a large-scale screening of PEA3
group member co-factors by using TAP-affinity methods (this
subject [PhD work of K. Verreman] will be described in the
project section).
Sumoylation
Post-translational modifications on lysines also play crucial
roles in the regulation of transcription, generally on histone
proteins but also on transcription factors, this is for example
the case of the acetylation of the PEA3 group members. These
last years, our research team has focused his interest on the
role of two linked post-translational modifications, i.e.
ubiquitinylation and sumoylation. In fact, both modifications
consist in the covalent addition of a polypeptide on a lysine
residue of the target protein. So, SUMO (Small Ubiquitin
related MOdifier) is a polypeptide of 97 residues, which is
generally linked as a monomer to a lysine being in the
consensus sequence ψKxE (ψ = hydrophobic residue).
Ubiquitin (Ub) consists of a polypeptide of 76 residues. The
SUMO and Ub pathway is composed of three similar
enzymatic activities: E1 (activation of SUMO and Ub), E2
(Ubc9) (conjugation to the target lysine, Ubc9 for SUMO), and
E3 ligase enzyme. For SUMO, three classes of E3 ligases have
been identified : PIAS, RanBP2 and Pc2 (Geiss-Friedlander &
Melchior, Nat Rev Mol Cell Biol 8: 947; 2007). Although the
ubiquitinylation process generally leads to protein
degradation, in most cases the sumoylation of a transcription
factor could reduce its ability to transactivate its target genes,
change its DNA binding capacity or modify the nucleocytoplasmic shuttle (Zhao, Cell Mol Life Sci 64: 3017; 2007).
Concerning the PEA3 group members, we have shown that
they are sumoylated on five consensus sites. This posttranslational modification clearly induces a drastic decrease
of the transcriptional activity of the group members (Degerny
et al., J Biol Chem 280 : 24330; 2005 - Degerny et al., BBA
1779 : 183; 2008). Our data also indicate that, in contrast to
what happens for another Ets protein (Yang & Sharrocks, Mol
Cell, 13: 611; 2004), the repression activity induced by
sumoylation of the PEA3 group is not due to the recruitment
of a Histone Deacetylase. However, the PIAS E3 ligases seem
to play a crucial role in this phenomenon. More specifically,
PIAS1 interacts with Erm to directly regulate its sumoylation
(PhD work of C. Degerny [2004-2007]).
Since 2004, our group has also collaborated on different research
programs linked to the transcriptional regulation :
- Yvan de Launoit was previously Director of a laboratory at
the ULB in Brussels where he initiated collaboration with
F. Fuks on epigenetic modifications in cancer. This study
permitted to prove that transcriptional repression could result
from a double lock: 1) DNA methylation and 2) histone
methylation or deacetylation. In parallel, we focused our
attention on the targeting of enzymes responsible for DNA
methylation, i.e. Dnmt, and we showed that the Myc
oncoprotein targets the Dnmts on DNA. Five papers were
published on this collaboration and the following since 2005
(Brenner et al., Embo J 26: 336; 2005 - Bernard et al., Oncogene
25: 1358; 2006; Viré et al., Nature 439: 871; 2006 - Burgers et al.,
Oncogene 26: 1650; 2007).
- We also collaborated with C. Van Lint at the ULB on the
transcriptional regulation of retroviruses such as HIV, HTLV and
BLV. Altogether, we characterized the different regions of the
LTR as well as intragenic regions responsible for the dramatic
increase of proviral transcription induced by the retroviral
protein Tat/Tax. Since 2002, we published 7 original papers on
this topic; 3 from 2004 (Nguyen et al., J Biol Chem 279: 35025;
2004 - Goffin et al., Nucleic Acids Res 33: 4285; 2005 – Nguyen
et al., J Biol Chem 282: 20854; 2007).
- Finally, Didier Monté completed the work related to the PhD
thesis of Z. Kherrouche on the identification of E2F target
genes (Kherrouche et al., BBRC 317: 749; 2004 – Kherrouche
et al., Biochem J 383: 52; 2004 – Kherrouche et al., Biochem
J 396: 547; 2006).
Publications
Ubiquitinylation
Concerning the second post-translational modifications
studied in our laboratory, we took advantage of our
observation that these proteins display a relatively short halflife to investigate ubiquitinylation. We have shown that these
transcription factors are submitted to this latter posttranslational modification, which is responsible for their
degradation through the 26S proteasome (Baert et al.,
Oncogene 26: 415; 2007). Concerning the E3 ligases involved
in PEA3 group member ubiquitinylation, we focused our
2007
Baert JL, Beaudoin C, Monté D, Degerny C, Mauen S, de Launoit, Y.
The 26S proteasome system degrades the ERM transcription factor and
regulates its transcription-enhancing activity.
Oncogene, 2007, 26:415-424.
Burgers WA, Blanchon L, Pradhan S, de Launoit Y, Kouzarides T, Fuks F.
Viral oncoproteins target the DNA methyltransferases.
Oncogene, 2007, 26:1650-1655.
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Cai C, Hsieh CL, Omwancha J, Zheng Z, Chen SY, Baert JL, Shemshedini L.
ETV1 is a novel androgen receptor-regulated gene that mediates prostate
cancer cell invasion.
Mol Endocrinol, 2007, 21:1835-1846.
Gosselin K, Martien S, Pourtier A, Vercamer C, Ostoich P, Morat L,
Sabatier L, Duprez L, T'kint de Roodenbeke C, Gilson E, Malaquin N,
Wernert N, Slijepcevic P, Ashtari M, Chelli F, Deruy E, Vandenbunder B,
De Launoit Y, Abbadie C.
Senescence-associated oxidative DNA damage promotes the generation
of neoplastic cells.
Cancer Res, 69:7917-7925.
Degerny C, Vintonenko N, Deheuninck J, Foveau B, Leroy C, Coll J,
Tulasne D. Baert JL, Fafeur V.
Regulation of the Ets-1 transcription factor by sumoylation and
ubiquitinylation.
Oncogene, 2007, 26:395-406.
Humbert N, Martien S, Augert A, Da Costa M, Mauen S, Abbadie C,
de Launoit Y, Gil J, Bernard D.
A genetic screen identifies topoisomerase 1 as a regulator of
senescence.
Cancer Res, 2009, 69:4101-4106.
Kondoh H, Lleonart M, Nakashima Y, Yokode M, Tanaka M,
Bernard D, Gil J, Beach, D.
A high glycolytic flux supports the proliferative potential of murine
Embryonic Stem cells.
Antiox Redox Signal, 2007, 9:293-299.
Reuse S, Calao M, Kabeya K, Guiguen A, Gatot JS, Quivy V, Vanhulle C,
Lamine A, Vaira D, Demonte D, Martinelli V, Veithen E, Cherrier T,
Avettand V, Poutrel S, Piette J, de Launoit Y, Moutschen M, Burny A,
Rouzioux C, De Wit S, Herbein G, Rohr O, Collette Y, Lambotte O,
Clumeck N, Van Lint C.
Synergistic activation of HIV-1 expression by deacetylase inhibitors and
prostatin : implications for treatment of latent infection.
PLoS One, 2009,4:e6093.
Nguyen TL, de Wlaque S, Veithen E, Dekoninck A, Martinelli V, de
Launoit Y, Burny A, Harrod R, van Lint C.
Transcriptional Regulation of the Bovine Leukemia Virus Promoter by the
Cyclic AMP-response Element Modulator {tau} Isoform.
J Biol Chem, 2007, 282:20854-20867.
Zdanov S, Bernard D, Debacq-Chainiaux F, Martien S, Vercamer C,
Gosselin K, Chelli F, Toussaint O, Abbadie C.
Normal or stress-induced fibroblast senescence involves COX-2 activity.
Exp Cell Res, 2007, 313:3046-3056.
Yockell-Lelièvre J, Spriet C, Cantin P, Malenfant P, Heliot L,
de Launoit Y, Audette M.
Functional cooperation between Stat-1 and ets-1 to optimize icam-1 gene
transcription.
Biochem Cell Biol, 2009, 87:905-918.
2008
2010
Acosta JC, O'Loghlen A, Banito A, Guijarro MV, Augert A, Raguz S,
Fumagalli M, Da Costa M, Brown, C, Popov N, Takatsu Y, Melamed J,
d'Adda di Fagagna F, Bernard D, Hernando E., Gil J.
Chemokine Signaling via the CXCR2 Receptor Reinforces Senescence.
Cell, 2008, 133:1006-1018.
Baert JL, Monté D, Verreman K, Degerny C, Coutte L, de Launoit Y.
The E3 ubiquitin ligase complex component COP1 regulates PEA3 group
member stability.
Oncogene, 2010, 29:1810-1820.
Dessein AF, Stechly L, Jonckheere N, Dumont P, Monté D, Leteurtre E,
Truant S, Pruvot FR, Figeac M, Hebbar M, Lecellier CH, Lesuffleur T,
Dessein R, Grard G, Dejonghe MJ, de Launoit Y, Furuichi Y, Prévost G,
Porchet N, Gespach C, Huet G.
Autocrine Induction of Invasive and Metastatic Phenotypes by the MIFCXCR4 Axis in Drug-Resistant Human Colon Cancer Cells.
Cancer Res, 2010, 70:4644-4654.
Degerny C, de Launoit Y, Baert JL.
ERM transcription factor contains an inhibitory domain which functions
in sumoylation dependent manner.
BBA, 2008, 1779:183-194.
Gouyer V, Fontaine D, Dumont P, de Wever O, Fontayne-Devaux H,
Leteurtre E, Truant S, Delacour D, Drobecq H, Kerckaert JP,
de Launoit Y, Bracke M, Gespach C, Desseyn JP, Huet G.
Autocrine induction of invasion and metastasis by Tumor-Associated
Trypsin Inhibitor (TATI) in human colon cancer cells.
Oncogene, 2008, 27:4024-4033.
Humbert N, Navaratnam N, Augert A, Da Costa M, Martien S, Wang J,
Martinez D, Abbadie C, Carling D, de Launoit Y, Gil J, Bernard D.
Regulation of ploidy and senescence by the AMPK-related kinase NUAK1.
EMBO J, 2010, 29:376-386.
Firlej V, Ladam F, Brysbaert G, Dumont P, de Launoit, Y, Benecke A,
Chotteau-Lelièvre A.
Reduced tumorigenesis in mouse mammary cancer cells following
inhibition of either of the distinct Pea3 and Erm transcription programs.
J Cell Sci, 2008,. 121:3393-3402.
Lens Z, Dewitte F, Monté D. Baert JL, Bompard C, Sénéchal M,
Van Lint C, de Launoit Y, Villeret V & Verger A.
Solution structure of the N-terminal transactivation domain of ERM
modified by SUMO-1.
BBRC, 2010, 399:104-110.
2009
PhD
Augert A, Bernard D.
Senescent-associated factors : pro- and anti-tumoral actions.
Med Sci, 2009, 25:789-790.
Augert A, Payré C, de Launoit Y, Gil J, Lambeau G, Bernard D. 2009
The M-type receptor PLA2R regulates senescence through the p53
pathway.
EMBO Rep, 2009, 10:271-277.
Cindy DEGERNY
Directeur : J.L. Baert
PhD « Biologie et Santé »
Octobre 2009
Gosselin K, Deruy E, Martien S, Vercamer C, Bouali F, Dujardin T,
Slomianny C, Houel-Renault L, Chelli F, De Launoit Y, Abbadie C.
2009
Senescent keratinocytes die by autophagic programmed cell death.
Am J Pathol, 2009, 174:423-435.
Kathye VERREMAN
Directeur : Y. de Launoit
PhD « Biologie-Santé »
Novembre 2009
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Cancer
The prostate cancer skeletal metastases are mostly
radiographically characterized as osteoblastic (i.e., increased
mineral density at the site of the lesion) as opposed to
osteolytic. The mechanisms through which prostate cancer
cells promote bone mineralization remain poorly
understood. Although many potentially responsible proteins
have been identified, to date, none has been shown to
contribute to prostate cancer–induced osteoblastic activity
in vivo in the context of prostate cancer. In this context, we
look for the Erg target genes expression associated with this
phenomenon. Two sources of biological materials will be
included in our study: firstly, biopsies of prostate cancers and
metastasis obtained from the Lille Tumor library of the CNO
(Cancerolope Nord-Ouest) ; secondly, prostate cancer cell
lines known to induce bone metastasis. Among the tested
cell lines, we will choose some of them to modulate the Erg
expression and study both their target genes expression and
their potential change in capacity to induce bone metastasis.
ETS Proteins, Transcriptional
Regulation and Associated
Diseases
Martique DUTERQUE-COQUILLAUD
CR1 - CNRS
The main approaches of our group aim to understand the
transcriptional properties of Ets transcription factors,
particularly Erg, and their target genes. Since in situ analyses
reveals the direct association of Erg expression with the
precartilaginous condensation and chondrogenesis
preceding bone formation, the Erg gene may be involved in
the early events of skeleton formation.
Identify the Erg target genes involved in cartilage
formation and maintenance
We have already obtained transgenic mice over-expressing
an Erg mutant protein, with trans-dominant negative effect,
specifically in cartilage under the Collagen2a1 promoter
control. The transgenic mice gradually developed earlyageing associated phenotypes including hyperlordosis/
hyperkyphosis, and reduced mobility. These results has been
confirmed by radiological and histological studies, and
suggest that the transgene expression affects the cartilage
formation, which allows premature erosion (HolderEspinasse el al., in preparation).
Our goal is to identify abnormal expressed genes associated
with the phenotype observed in transgenic mice expressing
the Erg dominant-negative protein. We chose a DNA-array
technology, which provides a powerful tool to obtain an
overview on gene expression patterns. We then analysed
and compared the transcriptome of chondrocytes freshly
isolated from wt and transgenic mice. We obtained a series
of genes and validate their up- or down regulation by
quantitative RT-PCR. A lot of studied genes were known to
be involved in cartilage formation or in cellular physiology.
Some of them were unknown. Among these genes, we
identified a new one giving rise to the UCMA protein (Upper
zone of growth plate and cartilage matrix associated protein)
described as a novel secretory protein mainly expressed in
cartilage and also as a novel vitamin K-dependent protein
named GRP (Gla-rich protein). This protein has the highest
Gla content of any protein known to date. We identify four
alternatively spliced variants of Ucma/GRP gene transcripts
associated with the early stages of chondrogenesis. They
give rise to four proteins, two of them are secreted and the
two others aggregate in cytoplasm. We suggest that the coexpression of soluble and insoluble isoforms (Gla-rich and
Gla-deleted isoforms) may be finely tuned. Imbalance in
isoform expression may therefore be involved in skeletal
pathology.
Publications
Define the link between the Erg gene over-expression
and bone metastasis of prostate cancers
Since prostate cancers have recently been associated with
Erg gene over-expression (more than 50% of the cases), we
will try to establish a link between Erg targets and prostate
cancers. Moreover, the most common site of prostate cancer
metastasis is the bone, with skeletal metastases identified at
autopsy in up to 90% of patients dying from prostate cancer.
Salentey V, Claus S, Bougault C, Paumier A, Aubert-Foucher E,
Ronzière M-C, Freyria A-M, Galera P, Beauchef G, DuterqueCoquillaud M, Piperno M, Damour O, Herbage B, Mallein-Gerin F.
Human chondrocyte responsiveness to Bone Morphogenetic Protein-2
after their in vitro dedifferentiation : potential use of Bone Morphogenetic
Protein-2 for cartilage cell therapy.
Pathol Biol, 2009, 57:282-289.
Moreover, in 2008 and 2009, our group collaborated on
different research programs linked to ETS proteins or
cartilage :
- with M. Aumercier, who participated to the team between
2006 and 2009, to study a novel ETS-1 isoform (Laitem et
al., 2009).
- with CH. Marquette, a pneumologist of Lille CHRU, to
contribute to tracheal replacement with aorta project
(Seguin et al., 2009, Markis et al., 2009). Since 2008, more
than ten patients have been operated with success.
- with F. Mallein-Gerin, IBCP Lyon (ANR-PROMOCART
project), to identify molecular mechanisms that control
the differentiated chondrocyte phenotype (Salentey et al.,
2009).
2009
Makris D, Holder-Espinasse M, WurtzA, Seguin A, Hubert T,
Jaillard S, Copin MC, Jashari R, Duterque-Coquillaud M, Martinod
E, Marquette CH.
Tracheal replacement with cryopreserved allogenic aorta.
Chest, 2009, 137:60-67.
Laitem C, Leprivier G, Choul-Li S, Begue A, Monte D, Larsimont D,
Duterque-Coquillaud M, Aumercier M.
Ets-1 p27: a novel Ets-1 isoform with dominant negative effects on the
transcriptional properties and the subcellular localization of Ets-1 p51.
Oncogene, 2009, 20:2087-2099.
Seguin A, Radu D, Holder-Espinasse M, Bruneval P, FialaireLegendre A, Duterque-Coquillaud M, CarpentierA, Martinod E.
Tracheal replacement with cryopreserved, decellularized, or
glutaraldehyde-treated aortic allografts.
Ann Thorac Surg, 2009, 87:861-867.
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Cancer
2010
Pathology and Immunotherapy of VirusAssociated Cancers
Le Jeune M, Tomavo N, Flourens A, Tian VT, Marchand N,
Camuzeaux B, Duterque-Coquillaud M.
Identification of four alternatively spliced transcripts of the Ucma/GRP
gene, encoding a new Gla-containing protein.
Exp Cell Res, 2010, 316:203-215.
Véronique PANCRE
CR1 - Inserm
Makris D, Holder-Espinasse M, Wurtz A, Seguin A, Hubert T,
Jaillard S, Copin MC, Jashari R, Duterque-Coquillaud M,
Martinod E, Marquette CH.
Tracheal replacement with cryopreserved allogenic aorta.
Chest, 2010, 137:60-67.
Our main subject focused on immunological events occurring
in tumoral pathologies associated with viral infections. We are
particularly interested in Hepatitis C virus (HCV) and Epstein
Barr virus (EBV) infections respectively described in major case
of human hepatocellular carcinoma and EBV associated
malignancies. In the context of these viral infections, we
analyzed both effector and regulatory T lymphocyte immune
responses during tumoral processes. We also are contributing
to immunotherapeutic protocol development, using peptide
presentation to effector targets, in order to promote
antitumoral immunity.
PhD
Marion LE JEUNE
Directeur : Martine Duterque-Coquillaud
PhD Biologie-Santé
Décembre 2009
The Epstein-Barr virus (EBV) program
The Epstein-Barr virus (EBV) is associated with several
malignant diseases which can be distinguished by their
patterns of viral latent gene expression. The latency II program
is limited to the expression of the non-immunodominant
antigens EBNA-1, LMP-1 and LMP-2 and is seen in EBV-positive
Hodgkin’s disease, nasopharyngeal carcinomas and peripheral
T/NK-cell lymphomas.
1. Immunotherapeutic strategy against EBV latency II
malignancy
CD4+ T cells may play a crucial role in controlling these EBV
latency II malignancies. We used the TEPITOPE software to
predict promiscuous MHC class II epitopes derived from the
latency II antigens EBNA-1, LMP-1 and LMP-2. The predicted
peptides were then submitted to peptide-binding assay on
HLA II purified molecules, which allowed the selection of 6
peptides (EBNA- 1: 3, LMP-1: 1, LMP-2: 2) with a highly
promiscuous capability of binding. The peptide cocktail is
highly immunogenic in HLA-DR1 transgenic mice, leading to
a specific cellular and humoral Th1 response. All the peptides
used in cocktail or individually were recognized by human
CD4+ T cells from healthy donors expressing various HLA II
genotypes and from patients with Hodgkin’s lymphoma (HL)
(Collaboration with Service des maladies du sang, CHRU, Lille).
EBNA 1-4 and LMP 2-4 peptides appear particularly dominants
in terms of proliferative activity and IFN- secretion.
Peptidespecific CD4 cell lines were able to lyse peptide-pulsed
lymphoblastoid cell lines (LCLs) or EBV expressing cell lines.
T cell repertoire analyses are currently under way. No
regulatory CD4 T cell activity is observed after peptide
stimulation. So, this promiscuous peptide cocktail could be
used in immunotherapeutic approaches (as peptide-based
vaccine or cellular therapy) against EBV latency II malignancies
so as to control the residual disease and to thus decrease the
risks of relapse. (Depil et al., Vaccine 24: 2225; 2006 - Depil et
al., J. Immunother 30: 215; 2007).
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Cancer
2. Identification of regulatory T cells in Hodgkin’s Lymphoma linked to EBV infection
In 20-40% of patients with HL, Epstein Barr Virus (EBV) is
present in the neoplastic cells, however very little is known
about the regulatory mechanisms induced in presence of EBV.
Using quantitative real time PCR, we observed in nodes of
EBV-positive HL patients, a significant increase of the
immunosuppressive cytokine IL-10 and of markers associated
with regulatory T cells (CD4+CD25+ and Tr1) when compared
with EBV-negative HL patients. This increase was confirmed
by immunohistochemical analysis on frozen node biopsies
and flow cytometric analysis on PBMCs of HL patients.
Moreover, we also described an over-expression of CCL17 and
CCL22 which attract Th2 and regulatory T cells and may evade
immune surveillance by Th1 cells and CTLs.
So, our study suggest the direct evidence of regulatory T cells
implication, particularly in EBV positive Hodgkin’s Lymphoma
and suggests a pivotal role of these cells in controlling the
antitumoral immune response in the context of viral infection.
represent a predictive factor of liver damage aggravation.
(Delhem, Cottrez et al., Bull Cancer, 95 : 1029; 2008).
- Identification and characterization of regulatory T cells
in recurrent hepatitis C after liver transplantation for
hepatocellular carcinoma linked to HCV infection.
To date, liver transplantation (LT) represents the unique
alternative to tumoral status. However, 80% of transplanted
patients show a severe viral infection recurrence, with
accelerated development of the HCC in 5 years. Our study
including 1 and 5 years after LT, stable HCV negative patients,
patients with minimal hepatitis C recurrence and patients with
severe hepatitis C recurrence, demonstrates clearly that
regulatory T cells (CD4+CD25+ and Tr1) and associated
cytokines (IL-10 and TGF- ) are significantly enhanced in
recurrent hepatitis C. In particular, Tr1 cells and IL-10 are
specifically enhanced in patients with severe recurrent
hepatitis C compare to stable HCV negative patients or
patients with minimal recurrence. So Tr1 cells could promote
the accelerated evolution by inhibition of the effective
antiviral response.
So, IL-10, 1 year after LT, could be a predictive marker of
recurrence progression. These findings may have therapeutic
implications since high IL-10 producers, one year after LT,
might represent a subset of liver transplant recipients which
require more intensive management. (Carpentier et al.,
Hepathology 44: suppl 1; 2006 – Carpentier et al., J Med Virol,
81 : 473; 2009 – Carpentier, Conti et al., Am J Transpl,9 : 2102;
2009).
The Hepatitis C Virus (HCV) Program
Hepatitis C virus (HCV) becomes chronic in about 85% of
infected individuals, whereas only 15% of infected people clear
spontaneously the virus. The progression of hepatitis C to
chronic status is associated to a profound down-regulation of
CD4 and CD8 multi-specific immune response. This immune
defect may participate to the immune tolerance of VHC and
consequently to its persistence. Recent findings indicate that T
regulatory cells as Tr1 play an inhibitory role on T helper
responses notably in the context of auto-immune or
inflammatory disorders. So, one hypothesis able to explain the
dysfunction of the immune response against HCV could be
immunosuppressive mechanisms supported by regulatory T
cells and particularly Tr1 cells via high interleukin-10 (IL-10)
secretion (Delhem, Carpentier et al., Bull Cancer 95: 1219; 2008).
- Inhibitory effect of Cyclosporine A (CsA) on human
regulatory T cells in vitro
HCV recurrence after liver transplantation represents a major
barrier to survival of the transplanted liver and may be
promoted by CD4+CD25+ regulatory T cells which are also
essential for transplant tolerance. Our results shows for the
first time that therapeutic doses of CsA, a widely used
immunosuppressive agent, did not change the phenotype
but significantly impaired the function of the
CD4+CD25+regulatory T cells by inducing pro-inflammatory
cytokines IL-2 and IFN- secretion.
So our study suggested that CsA may block the induction of
immune tolerance and decrease the risk of hepatitis C
recurrence (Miroux, Moralès et al., Transpl Proc, 41 : 3371 ; 2009).
In 2006, with our implication in the Cancer Program developed
by the UMR 8161, we decided to focus our activities on cancer
projects. In this sense, our project of peptide-based
immunotherapy of HIV infection was not carried out.
- Role of regulatory T cells in the fibrosis progression to
hepatocellular carcinoma
We have evaluated the existence of regulatory T cells in liver
biopsies of three well-defined cohorts of chronically HCV-1b
infected patients, including patients without liver lesions,
patients with cirrhosis and patients with hepatocellular
carcinoma (HCC) (Collaboration with the Service de
transplantation hépatique, Hôpital Cochin, Paris and with the
“Service de transplantation, Maladies de l’appareil digestif et
nutrition”, CHRU, Lille). Using quantitative real time PCR, we
demonstrated an increased expression of IL-10 and
transforming growth factor-beta (TGF- in liver biopsies with
more severe fibrosis. This observation was correlated with
increased expression of the specific markers of the CD4+CD25+
cells (FoxP3, GITR, CTLA-4, ICAM-1, P-selectin) and of the Tr1 cells
(Integrin 2, Integrin 2) during the pathogenesis progression.
Experiments performed on liver biopsies of a 15 year
HCVgenotype 1b infected patient, followed from the chronic
stage to the HCC, confirmed an increase of Tr1 proportionally
to the aggravation of the pathology. In contrast, we did not
detect Tr1 cells in liver biopsies from alcoholic cirrhosis and from
alcohol induced-hepatocellular carcinoma, evidencing a virusrelated presence of Tr1 cells.
So, the evidence of the presence of regulatory T cells in the liver
of chronically infected patients and the correlation with HCC
suggests a central role of these cells in HCV persistence and in
the evolution of the liver pathology and thus potentially
The Human Immunodeficiency Virus (HIV) Program
We previously identified the Nef 56-68 peptide as the first HIV
derived HLA-DQ-restricted peptide capable to induce HIVspecific memory CD4+ T cells producing IFN- in all the healthy
donors tested. The ex vivo response to this peptide used in
combination with three HLA-DR-restricted Nef peptides (Nef
66-97, Nef 133-159, Nef 180-202), identified with the TEPITOPE
Software, was evaluated in HIV-seropositive patients and Long
term Non Progressors (LTNPs). Analyzing specific proliferative
response and IFN- secretion, patients were identified as high
responders, medium responders and non responders to
peptides. As high responder patients, LTNP patients showed
strong proliferative response to all the Nefpeptides as strong
IFN- secretion. 24 months later, all high responder patients
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Cancer
Delhem N, Carpentier A, Moralès O, Miroux C, Groux H, Auriault C,
Pancré V.
Regulatory T Cells and Hepatocellular Carcinoma: Implication of the
Regulatory T lymphocytes in the control of the Immune Response.
Bull Cancer, 2008, 95:1219-1225.
and LTNPs were always without antiretroviral treatment
whereas 50% of medium responders and at least 66 % of low
responder patients followed bitherapy.
CDC classification confirmed also unfavourable evolution for
these two last groups. All high responder patients conserved
stable CD4 counts, proliferative response to Nef peptides as
strong IFN- secretion during this 24 month period.
So, early full T CD4 response to peptides of the Nef protein
could thus be regarded as a factor of good prognosis in HIV
infection and a tool of importance in the decision to put or
not a patient under treatment. These 4 Nef peptides also
constitute attractive components for a multiepitopic vaccine
toward HIV infection (Pancré et al., Vaccine 25: 5927; 2007).
Delhem N, Cottrez F, Carpentier A, François V, Moralès O, Miroux C,
Groux H, Auriault C, Pancré V.
Role of the Regulatory T lymphocytes in hepatitis C fibrosis progression.
Bull Cancer, 2008, 95:1029-1038.
Kitajima K, Taboury J, Boleslawski E, Savier E, Vaillant JC, Hannoun L.
Sonographic preoperative assessment of liver volume before major liver
resection.
Gastroenterol Clin Biol, 2008, 32:382-389.
2009
Publications
Carpentier, Carrière M, Aoudjehane L, Miroux C, Moralès O, Conti F,
Calmus Y, Chaussade S, Groux H, Auriault C, Pancré V, Delhem N,
Podevin P.
Analysis of gene transcription in sera during chronic hepatitis C.
J Med Virol, 2009, 81:473-480.
2007
Depil S, Moralès O, Maillère B, Delhem N, François V, Georges B,
Dufossé F, Morschhauser F, Hammer J, Auriault C, Pancré V.
Determination of a HLA II promiscuous peptide cocktail as potential
vaccine against EBV latency II malignancies.
J Immunotherapy, 2007, 30:215-226.
Carpentier A, Conti F, Stenard F, Aoudjehane L, Miroux C, Podevin P,
Morales O, Chouzenoux S, Scatton O, Groux H, Auriault C, Calmus Y,
Pancre V, Delhem N.
Increase expression of regulatory Tr1 cells in recurrent hepatitis C after
liver transplantation.
Am J Transplant, 2009, 9:2102-2112.
Dharancy S, Boitard J, Decaens T, Sergent G, Boleslawski E, Duvoux C,
Vanlemmens C, Meyer C, Gugenheim J, Durant F, Boillot O, Declerck
N, Louvet A, Canva V, Romano O, Ernst S, Mathurin P, Pruvot FR.
Comparison of two techniques of transarterial chemoembolization
before liver transplantation for hepatocellular carcinoma: a case-control
study.
Liver Transpl, 2007, 13:665-671.
Miroux C, Moralès O, Carpentier A, Dharancy S, Conti F, Boleslowski E,
Podevin P, Auriault C, Pancré V, Delhem N.
Inhibitory effectos of cyclosporine on human regulatory cells in vitro.
Transplant Proc, 2009, 41:3371-3374.
2010
Lerut JP, Orlando G, Adam R, Schiavo M, Klempnauer J, Mirza D,
Boleslawsi E, Burroughs A, Sellés CF, Jaeck D, Pfitzamm R,
Salizzoni M, Söderdahl G, Steininger R, Wettergren A, Mazzaferro V,
Le Treut YP.
The place of liver transplantation in the treatment of hepatic epitheloid
hemangioendothelioma : report of the European liver transplant register.
Ann Surg, 2007, 246:949-957.
Chang Y, de Nadai P, Azzaoui I, Morales O, Delhem N, Vorng H,
Tomavo S, Ait Yahia S, Zhang G, Wallaert B, Chenivesse C,
Tsicopoulos A.
The chemokine CCL18 generates adaptive regulatory T cells from memory
CD4+ T cells of healthy but not allergic subjects.
FASEB J, 2010, Aug 11.
Louvet, Naveau S, Abdelnour M, Ramond MJ, Diaz E, Fartoux L,
Dharancy S, Texier F, Hollebecque A, Serfaty L, Boleslawski E,
Deltenre P, Canva V, Pruvot FR, Mathurin P.
The Lille model: a new tool for therapeutic strategy in patients with severe
alcoholic hepatitis treated with steroids.
Hepatology, 2007, 45:1348-1354.
PhD
Arnaud CARPENTIER
Co-Directeur : N.Delhem
PhD « Biologie, Physiologie »
Paris VI
Novembre 2009
Pancré V, Delhem N, Yazdanpanah Y, Delanoye A, Delacre M,
Depil S, Moralès O, Mouton Y, Auriault C.
Presence of HIV-1 Nef specific CD4 T cell response is associated with non
progression in HIV-1 infection.
Vaccine, 2007, 25:5927-5937.
Olivier MORALES
EPHE
Decembre 2009
2008
Boleslawski E, BenOthman S, Grabar S, Correia L, Podevin P,
Chouzenoux S, Soubrane O, Calmus Y, Conti F.
CD25, CD28 and CD38 expression in peripheral blood lymphocytes as a
tool for predicting acute rejection after liver transplantation.
Clin Transpl, 2008, 22:494-501.
HDR
Nadira DELHEM
HDR
Novembre 2009
Chang Y, Chenivesse C, Gilet J, Morales O, Delhem N, Legendre B,
Zhang G, Tsicopoulos A.
The chemokine CCL18 turns memory CD4+ T cells from non-allergic
subjects into T cells expressing a regulatory phenotype.
Rev Mal Respir, 2008, 25:1154.
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Cancer
investigated and characterised as explained below. Some of
the others were validated but are yet to be characterised. This
characterisation will constitute a part of the future research
project.
Identification and
Characterisation of New
Genetic Event Involved
in an early Step of Tumor
Development :
the Senescence Escape
1/ Topoisomerase I (Top1)
A manuscript concerning the characterisation of the role of
Top1 on senescence is currently in preparation (Humbert et
al. 1st paper). In this study, we report that knockdown of
topoisomerase I (Top1) results in an increased replicative
potential associated with a decrease in senescence markers
and a diminished DNA damage response. Conversely, Top1
constitutive expression induces growth arrest, the appearance
of a senescence marker, and activation of the DNA damage
response. Interestingly, double-strand breaks caused by
reactive oxygen species treatment trigger recruitment of Top1
to DNA. This recruitment has a strong impact on the
magnitude of the DNA damage response and on subsequent
cell growth arrest. Altogether, these results reveal an
unanticipated function of Top1 in regulating senescence and
the DNA damage response.
David BERNARD
CR2 - CNRS
Numerous genetic alterations are involved in cancer
development. These alterations lead to an activation of protooncogenes and a suppression of tumor-suppressor genes, to
confer new biological properties to the cancer cells (Hanahan
et al., Cell 100: 57; 2000). Thus, during the transformation
process and due to these genetic modifications, a normal cell
acquires new properties.
A new property acquired by tumor cells is the immortality and
the ability to be resistant to senescence induction. Senescence
was first defined in primary human fibroblasts after the cells
had reached their replicative potential and is characterised as
a permanent form of cell cycle arrest. In fact, cells also enter
senescence when they are stressed by various stimuli such as
oxidative or oncogenic stresses (Serrano et al., Curr Opin Cell
Biol 13: 748; 2001). Although senescent cells are mostly found
blocked in the cell cycle, they remain metabolically active and
display characteristic changes in their gene expression and
morphology (Yaswen et al., Cell 128: 233; 2007). They are
flattened, enlarged, and exhibit positive senescence
associated β galactosidase activity (Dimri et al., PNAS USA 92:
9363; 1995) and senescence associated heterochromatin foci
(Narita et al., Cell 113: 703; 2003).
A number of recent investigations show that senescence is a
tumor-suppression mechanism which is activated in the early
stages of tumorigenesis in order to prevent malignant
transformation (Braig et al., Nature 436: 660; 2005 - Chen et al.,
Nature 436: 725; 2005 - Collado et al., Nature 436: 642; 2005 Michaloglou et al., Nature 436: 720; 2005). Based on this idea,
an escape or dysfunction of the senescence program would
lead to a progression of malignancy. It therefore appears
necessary to understand why a cell under different stresses is
going to enter a senescent state and what genetic events
might impede this phenomenon.
Our research goal is to understand the genetic events
regulating senescence bypass. We started to lead a project in
2006 in the UMR 8161 on this topic. As a first approach, we
have performed a genetic screening using a retroviral shRNA
library in order to identify new genes regulating senescence
occurrence. HDFs displaying colonies, thus bypassing
senescence, were amplified before gDNA extraction and
subsequent shRNA identification. As a result from the
screening we have identified a few candidates controlling
senescence occurrence.
2/ ARK5
A draft reporting the characterisation of ARK5 on senescence
is currently in preparation (Humbert et al., 2nd paper). In this
work, we show that the down-regulation of the AMPK-related
protein kinase 5 (ARK5 or NUAK1) results in senescence
bypass. Interestingly, ARK5 expression increases during
replicative senescence and its ectopic expression triggers a
premature senescence. Our data suggest that ARK5 acts
mainly by blocking cytokinesis and inducing genomic
instability. Indeed, ARK5 down-regulates a known cytokinetic
regulator LATS1 and LATS1 inactivation mimics ARK5 effect
over senescence. ARK5 phosphorylates LATS1 on the Ser 464
and mutation of this residue is sufficient to reverse ARK5 effect
on LATS1 level. Collectively, these data suggest that an
enforced genomic stability, due to a down-regulation of ARK5,
can extend lifespan of normal human cells.
Major results since 2006
We have already identified and validated few genes involved
in senescence occurrence. The first two genes identified were
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Cancer
ES cells and display additional repressive marks in their
malignant counterpart the embryonic teratocarcinoma (EC)
cells. These studies strongly support the stem-cell origin of
cancer and help to understand how normal stem cells with a
transient transcription–ready state are converted into cancer
progenitor cells with the permanent and heritable epigenetic
silencing via DNA hypermethylation of some key genes
probably due to aberrant crosstalk between components of
Polycomb complexes and DNA-methyl transferases (DNMTs)
(Ohm et al., Nature Genet 39: 237; 2007). In fact, the loss of
function of these genes could draw these progenitor cells
from the normal differentiation pathway and lock them into
a state of perpetual self renewal poised for tumorigenesis with
the accumulation of further epigenetic or genetic events.
Thus, in early tumorigenesis, epigenetic inactivation of HIC1
could induce an increase in SIRT1 deacetylase activity. The
resulting deacetylation of P53 impairs its function leading to
a defective apoptotic response to DNA-damage and a reduced
ability to transactivate its target genes including HIC1 (Chen
et al., Cell 123: 437; 2005).
SIRT1 can deacetylate not only P53 but also many other target
proteins essential for normal homeostasis. Thus, epigenetic
silencing of HIC1 can have both P53-dependent and P53independent effects.
Functionnal Studies
on the Tumor
Suppression Gene HIC1
(Hypermethylated
in Cancer 1)
Dominique LEPRINCE
DR2 - CNRS
Group Members :
Loison Ingrid, Assistant Engineer CNRS
Dehennaut Vanessa, Postdoctoral fellow CNRS/ARC
Foveau Benedicte, Postdoctoral fellow CNRS/AICR
Boulay Gaylor, PhD Student CNRS/ARC
Guerardel Catheline, Technician IPL
HIC1, a tumor suppressor gene involved in a feedback
regulatory loop with p53 and SIRT1
HIC1 (Hypermethylated In Cancer 1) codes for a transcriptional
repressor and is located in 17p13.3, a region frequently
hypermethylated or deleted in human tumours (Wales et al.,
Nature Medicine 1: 570; 1995). HIC1 has been considered as a
candidate tumour suppressor since its enforced expression by
stable transfection in various cancer cell lines results in a
significant decrease in their clonogenic survival and since its
expression is up regulated by P53 (Wales et al., Nature
Medicine 1: 570; 1995 - Guérardel et al., J Biol Chem 276 : 3078;
2001).
Definitive clues to the tumour suppressor function of HIC1
have come from animal models developed in Dr Steve Baylin’s
laboratory. Heterozygous Hic1+/- mice develop a late-onset
and gender-specific spectrum of spontaneous tumours (Chen
et al., Nature Genet 33: 197; 2003). In addition, using Hic1 and
p53 double heterozygous knockout mice, it has been shown
that Hic1 cooperates with p53 in determining cancer
progression and spectrum (Chen et al., Cancer Cell 6: 387;
2004). Finally, a circular regulatory loop has been recently
proposed for HIC1, SIRT1 and p53. HIC1 directly represses the
transcription of SIRT1 through a SIRT1/HIC1 (Chen et al., Cell
123: 437; 2005) or a CtBP/HIC1 (Zhang et al., PNAS 104: 829;
2007) repressor complex. SIRT1 is a class III NAD+ dependant
deacetylase which deacetylates many protein targets,
including p53. Thus, SIRT1 negatively modulates P53 DNAbinding properties and hence transactivation of its direct
target genes, including HIC1 (Chen et al., Cell 123: 437; 2005;
Britschgi et al., Oncogene 25: 2030; 2006).
Recently, we have shown that HIC1 is a target of the class III
deacetylase SIRT1 and we have thus identified a new posttranslational modification step in the P53-HIC1-SIRT1
regulatory loop (Stankovic-Valentin et al., Mol Cell Biol 27:
2661; 2007 - Reviewed in Fleuriel et al., IJBCB In press; 2008).
In ES cells, the promoters of some developmental
transcription factors are in a “bivalent state” with both
activating and repressive epigenetic marks. By contrast, HIC1
as a high percentage of genes specifically hypermethylated in
adult cancers are already pre-marked with repressive marks in
HIC1 : a transcriptional repressor belonging to the
BTB/POZ family
During the last ten years, we have performed functional
studies on HIC1 and defined the HIC1 protein as a sequence
specific transcriptional repressor containing three main
functional domains: (i) the N-terminal BTB/POZ (which stands
for Broad complex, Tramtrack and Bric a brac/Poxviruses and
Zinc finger) domain of about 120 amino-acid is a dimerization
domain known to play direct or indirect (through
conformational effects) roles in protein-protein interactions.
This domain is also an autonomous transcriptional repression
domain; (ii) the C-terminal end contains five Krüppel-like C2H2
zinc fingers which bind a specific DNA sequence, termed the
HiRE (HIC1 responsive element) that we have defined (Pinte
et al., J Biol Chem 279: 38313; 2004) and a C-terminal tail that
displays no obvious functional domain but has been
phylogenetically conserved (Bertrand et al., BBA 1678: 57;
2004); (iii) a central region which is a second autonomous
transcriptional repression domain (Deltour et al., Mol Cell Biol
22: 4890; 2002 - Stankovic-Valentin et al., FEBS J 273: 2879;
2006).
The BTB/POZ domain is a highly conserved and widely
distributed structural motif found mainly in transcription
factors. BTB/POZ domains are protein-protein interaction
domains and mediate homo-oligomerization, heterooligomerization as well as interactions with non-BTB/POZ
proteins (Stogios et al., Genome Biol 6: R82; 2005). These
properties are essential for the biological function of BTB/POZcontaining proteins. We have shown that he HIC1 BTB/POZ
domain is also an autonomous transcriptional repression
domains but is insensitive to trichostatin A (TSA), a specific
inhibitor of class I and class II HDACs (Deltour et al., PNAS 96:
14831; 1999; Deltour et al., BBRC 287: 427; 2001). It has been
recently shown that HIC1 forms a transcriptional repression
complex with the class III HDAC, SIRT1 and that this complex
directly binds the SIRT1 promoter to repress its transcription
(Chen et al., Cell 123: 437; 2005). SIRT1 is a NAD-dependent
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Cancer
involved in the repression mediated by the isolated BTB/POZ
domain but by deacetylating HIC1, SIRT1 favors its
SUMOylation and thus the establishment of an optimal
transcriptional repression. In addition, they bring new insights
into the HIC1, P53 and SIRT1 regulatory loop. This loop relies
not only on direct transcriptional effects since P53 activates
the HIC1 promoter and HIC1 represses the SIRT1 promoter but
also on post-translational effects.
In conclusion, the HIC1 central region is essential for the
transcriptional repression potential of HIC1. But, corepressors
and complexes interacting with this region, notably those
whose recruitment is regulated by the SUMOylation/
Acetylation switch have still to be characterized.
The C-terminal end contains a cluster of four conserved C2H2
zinc fingers (ZF 2-5) which are separated by the typical 7-8
amino acid conserved H/C links found in Krüppel-like Zinc
fingers, making them likely to be involved in sequencespecific DNA binding. Using a combination of functional
assays and in silico analyses, we have identified the sequence
5’-C/GNGC/GGGGCAC/ACC-3’ as an optimal HIC1 binding site
(HiRE for HIC1-responsive element) and validated by
mutational analyses a GGCA core motif bound by Zinc fingers
3 and 4 (Pinte et al., J Biol Chem 279: 38313; 2004). The fulllength HIC1 protein is unable to bind to a single optimized site
and this DNA-binding inhibition is clearly BTB/POZdependent. Whereas the HIC1 BTB/POZ domain impedes
binding to a single site, it mediates strong cooperative binding
to a probe containing multiple optimized sites (Pinte et al., J
Biol Chem 279: 38313; 2004).
The first bona fide HIC1 direct target gene, SIRT1, contains a
cluster of two HiRE in the same orientation located at the 5’
end of the promoter (Chen et al., Cell 123: 437; 2005). ChIP and
ChIP upon ChIP assays demonstrated that HIC1 forms a
transcriptional repression complex with the deacetylase SIRT1
to directly bind the SIRT1 promoter and repress its
transcription.
deacetylase which is insensitive to TSA but sensitive to
nicotinamide (NIA). The HIC1 BTB/POZ domain interacts with
SIRT1 (Chen et al., Cell 123: 437; 2005) but its repressive activity
in the context of a Gal4 chimera is not inhibited by NIA (our
unpublished results).
Thus, the repression mechanisms inherent to the HIC1
BTB/POZ domain have still to be deciphered.
The HIC1 central region is also an autonomous transcriptional
repression domain but sensitive to TSA (Deltour et al., Mol Cell
Biol 22: 4890; 2002). This region has not been subjected to a
strong selection pressure except for 4 peptidic motifs perfectly
conserved from human to zebrafish (Bertrand et al., BBA 1678:
57; 2004). One of them, GLDLSKK, is highly related to the
canonical motif PxDLSxK/R found in proteins interacting with
the co-repressor CtBP (C-terminal Binding Protein). We have
shown that HIC1 interacts with the two related CtBP1 and
CtBP2 corepressors through this conserved GLDLSKK motif
thus extending the CtBP binding site (Deltour et al., Mol Cell
Biol 22: 4890; 2002) (Stankovic-Valentin et al., FEBS J 273: 2879;
2006). Notably, mutation of the central leucine residue, Leu
225 in HIC1, which is the only invariant residue in CtBPinteraction motifs, abolished the interaction between HIC1
and CtBP (Stankovic-Valentin et al., FEBS J 273: 2879; 2006). As
would be expected from the corepressor activity of CtBP, the
L225A point mutation or the deletion of the GLDLSKK motif
impairs the repression potential of the HIC1 central region, but
does not fully abolish it (Stankovic-Valentin et al., FEBS J 273:
2879; 2006). Thus, the HIC1 central region appears to be a
second repression domain exhibiting both CtBP-dependent
and CtBP-independent repression mechanisms, both being
sensitive to TSA.
The second conserved motif is an YRWM/VK314xEP motif
(Stankovic-Valentin et al., Mol Cell Biol 27: 2661; 2007) which
contains a sumoylation consensus site, KxE. Sumoylation is a
reversible post-translational modification in which a member
of the Small Ubiquitin-like Modifier (SUMO) family of proteins
is covalently conjugated to lysine residues in target proteins.
Unlike ubiquitination which generally marks proteins for rapid
degradation, the SUMOylation of nuclear proteins has very
diverse effects on transcriptional activity ranging from
regulation of DNA-binding activity, subcellular localization
and assembly of multiprotein complexes. In the case of HIC1,
SUMOylation of K314 does not affect its subnuclear
localization and its interaction with CtBP, HDAC4 and SIRT1
but does positively regulate the transcriptional repression
potential (Stankovic-Valentin et al., Mol Cell Biol 27: 2661;
2007).
Lysine residues can be targeted by several post-translational
modifications including SUMOylation, acetylation, ubiquitination or methylation. The KxEP motif in HIC1 with the Proline
residue conserved from human to zebrafish is related to the
G/SKxxP consensus motif for acetylation by CBP/P300. Indeed,
we have shown that HIC1 is acetylated on various Lysine
residues including K314. Thus, the KxEP motif is an
acetylation/SUMOylation switch motif. The cross-talk between
these two competitive post-translational modifications of
Lysine residues is orchestrated by a new complex associating
two distinct types of deacetylases, HDAC4 and SIRT1. Even
though the precise mechanisms are not fully understood,
these results identify HIC1 as a new target of the class III
deacetylase SIRT1. Our results thus provide the first
mechanistic clues to the HIC1/SIRT1 interaction. SIRT1 is not
The chemokine receptor CXCR7 is a direct HIC1 target gene
To identify the direct target genes of HIC1, we performed
microarray gene expression profiling analyses using the
Affymetrix U133A gene chip and total RNAs from U2OS cells
infected by adenovirus-expressing HIC1 or GFP as control, in
collaboration with Dr Brian Rood (CNMC, Washington). We
have used RNAs extracted during a time course infection
(from 8 to 26 Hours), the rationale behind being that the
earliest repressed genes are more likely to be the direct targets
of HIC1. Indeed, we identified a total of 94 genes whose
expression was down-regulated at least 3-fold. Some of these
genes are involved in proliferation, apoptosis and/or cell cycle
control with no obvious “clustering”.
We have first focused our studies on CXCR7/RDC1, an orphan
G protein coupled receptor shown to be a second receptor, in
addition to CXCR4, for the chemokine SDF-1 (stromal cellderived factor 1)/CXCL12. The SDF-1/CXCR4 chemotactic
pathway is a key player in the crosstalk between various tumor
cells and their microenvironment. First, CXCR4 is essential for
metastatic spread to organs where CXCL12 is expressed.
Second, stromal fibroblast-derived CXCL12 can stimulate
survival and growth of neoplastic breast cells and can
promote tumor angiogenesis by recruiting circulating
endothelial cells to the tumor microenvironment. Similarly,
upon ectopic expression of CXCR7, NIH3T3 become
tumorigenic in nude mice. CXCR7 is highly expressed in
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Cancer
Jenal M, Trinh E, Britschgi C, Britschgi A, Roh V, Vorburger SA,
Tobler A, Leprince D, Fey MF, Helin K, Tschan MP
The tumor suppressor gene hypermethylated in cancer 1 is
transcriptionally regulated by E2F1.
Mol Cancer Res, 2009, 7:916-922.
human primary breast, prostate and lung tumors and it is
expressed in both malignant cells and in the tumor-associated
vasculature but not in normal blood vessels. Finally, inhibition
of CXCR7 by siRNA interference or by a specific high affinity
small molecule antagonist severely reduces in vivo tumor
growth in animal models.
The use of qRT-PCR analyses and luciferase reporter assays,
CXCR7 has been validated as an HIC1 target gene in U2OS cells
overexpressing HIC1. Moreover, ChIP experiments
demonstrated that endogenous HIC1 proteins are bound to
the CXCR7 and SIRT1 promoters in normal human fibroblast
WI38 cells and neuroblastoma SKNMC cells.
We have set up an alternative experimental approach to
characterize HIC1 target by inhibiting its repressive function
through a “dominant-negative” strategy. We have thus,
constructed a dominant-negative version of HIC1 by cloning
the isolated Zinc finger domain downstream of a FLAG
epitope in a modified pcDNA3 expression vector. This HIC1 DN
protein will bind with very high affinity to single HiRE,
compete for the binding of the endogenous HIC1 proteins to
their target genes and hence counteract their repression
(Pinte et al., J Biol Chem 279: 38313; 2004). As a model, we
have used the hTERT-HME1 cell line (Clontech) which are
Telomerase-immortalized normal human breast epithelial
cells expressing HIC1. The U2OS cells used in the adenovirusbased experiment are not suitable since as most transformed
cell lines, they don’t express HIC1. Upon stable transfection of
this dominant negative (DN) expression vector into hTERTHME1, two clones resistant to G418 have been selected.
Surprisingly, these cells do not have a growth advantage in
monolayer but display a transforming and invasive phenotype
as shown by anchorage-independent cell growth, wound and
Transwell invasion assays). In addition and in striking contrast
with the parental hTERT cells, these DN cells induce tumours
in SCID mice. Taken together, our results implicate the tumor
suppressor HIC1 in the transcriptional regulation of the
chemokine receptor CXCR7 which plays key functions in the
promotion of tumorigenesis in a wide variety of cell type (Van
Rechem et al., JBC, 2009).
Van Rechem C, Boulay G, Leprince D
HIC1 interacts with a specific suunit of SWI/SNF complexes,
ARID1A/BAF250A.
Van Rechem C, Rood BR, Touka M, Pinte S, Jenal M, Guérardel C,
Ramsey K, Monté D, Bégue A, Tschan MP, Stephan DA, Leprince D.
Scavenger chemokine (CXC motif) receptor 7 (CXCR7) is a direct target
gene of HIC1 (hypermethylated in cancer 1).
J Biol Chem, 2009, 284:20927-20935.
Zhang B, Chambers KJ, Leprince D, Faller DV, Wang S.
Requirement for chromatin-remodeling complex in novel tumor
suppressor HIC1-mediated transcriptional repression and growth control.
Oncogene, 2009, 28:651-661.
Dehennaut V, Leprince D
Implication of HIC1 (Hypermethylated in Cancer 1) in the DNA damage
response.
Bulletin du Cancer, 2009, 96:E66-E72. Review.
2010
Guinez C, Mir AM, Martin N, Leprince D, Michalski JC, Vergoten G,
Lefebvre T.
Arginine 469 is a pivotal residue for the Hsc70-GicNAc-binding property.
Van Rechem C, Boulay G, Pinte S, Stankovic-valentin N, Guerardel C,
Leprince D.
Differential regulation of HIC1 target genes by CtBP and NuRD via an
acetylation/SUMOylation switch, in quiescent versus proliferating cells.
Mol Cell Biol, 2010, 30:4045-4059.
PhD
Capucine Van RECHEM
Directeur : D Leprince
Septembre 2009
Publications
2007
Stankovic-Valentin N, Deltour S, Seeler J, Pinte S, Vergoten G,
Guerardel C, Dejean A, Leprince D.
An Acetylation/Deacetylation-SUMOylation switch through a
phylogenetically conserved KxEP motif in the tumor suppressor HIC1
(Hypermethylated in Cancer 1) regulates its transcriptional activity.
Mol Cell Biol, 2007, 27:2661-2675
Zhang Q, Wang SY, Fleuriel C, Leprince D, Rocheleau JV, Piston DW,
Goodman RH.
Metabolic regulation of SIRT1 transcription via a HIC1: CtBP co-repressor
complex.
2009
Van Rechem C, Touka M, Boulay G, Guerardel C, Rood BR,
Leprince D.
HIC1 (Hypermethylated in Cancer 1) epigenetic silencing in tumors.
Int J Biochem Cell Biol, 2009, 41:26-33.
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Cancer
produce chemotactic, growth, and survival factors for mural
cells, such as platelet-derived growth factor (PDGF, (Zerwes &
Risau, J Cell Biol 105: 2037; 1987) and fibroblast growth factors
(Vlodavsky et al., Proc Natl Acad Sci USA 84: 2292; 1987 Schweigerer et al., Nature 325: 257; 1987) which recruit the
SMC to the newly formed vessel (Conway et al., Cardiovasc Res
49: 507; 2001). Mural cells produce endothelial growth and
chemotactic factors, such as VEGF (Ferrara et al., Growth
Factors 5: 141; 1991) and fibroblast growth factor-2 (Winkles
et al., Proc Natl Acad Sci USA 84: 7124 ; 1987), which participate
to the initial formation and progression of the capillary as well
as its maturation. Other factors, such as the angiopoietins and
their Tie-2 receptor (Davis et al., Cell 87: 1161; 1996 Maisonpierre et al., Science 277: 55; 1997 - Witzenbichler et al.,
J Biol Chem 273: 18514; 1998) or transforming growth factorβ (Orlidge & D'Amore, J Cell Biol 105: 1455; 1987 - Sato & Rifkin,
J Cell Biol 109: 309; 1989 - Sato et al., J Cell Biol 111: 757; 1990
- Sacchi et al., Oncogene 6: 2149; 1991) participate to blood
vessel maturation and stabilization. Direct contacts between
endothelial and mural cells as well as with the extra-cellular
matrix also take part to the dialog.
The formation of a locally stable and functional vascular tree
ultimately depends on all these complex interactions. Major
pathological incidences are the result of a perturbation of
these exchanges; in arteriosclerosis, SMC migration and
proliferation and a damaged endothelium are key factors for
the progression of the initial lesion (Behrendt & Ganz, Am J
Cardiol 90: 40L; 2002). In solid tumors, the formation of an
irregular and poorly structured blood vessel network is
partially the result of a lack of coordinated interactions
between these cells (Carmeliet & Jain, Nature 407: 249; 2000).
Interestingly, the so-called ‘anti-angiogenic therapies’ have
little to no clinical beneficial effects when used alone.
However, they appear to potentiate the classical
chemotherapies which they are combined with. It is now clear
that the ‘anti-angiogenic’ compounds do not induce tumor
asphyxia; they rather seem to normalize the vessels, allowing
a better access of chemotherapeutic compounds to the tumor
mass (Jain, Science 307: 58; 2000). This normalization is highly
dependent on the above described molecular dialog that
takes place between endothelial and perivascular cells in
order to form stable, functional vessels as those formed during
normal angiogenesis.
We have identified a new gene, VE-statin (also later named
egfl7), which is specifically expressed by endothelial cells of
the developing mouse embryo and in the adult, and in early
endothelial progenitors (Soncin et al., Embo J 22: 5700; 2003).
Analysis of VE-statin/egfl7 expression during embryogenesis
revealed that VE-statin/egfl7 is present in blood islands, the
first organized vascular structures in the embryo. Expression
is maintained throughout embryonic development in all
endothelial cells, whereas it declines in adults (Soncin et al.,
Embo J 22: 5700; 2003 - Campagnolo et al., Am J Pathol 167:
275; 2005 - Fitch et al., Dev Dyn 230: 316; 2004 - Parker et al.,
Nature 428 : 754 ; 2004). Expression is up-regulated in
physiological and pathological processes such as pregnancy,
tumor growth or after arterial denudation Soncin et al., Embo
J 22: 5700; 2003 - Parker et al., Nature 428: 754; 2004).
VE-statin/egfl7 codes for a 30 kDa protein composed of a signal
sequence, an EMI domain (previously described in emilin and
involved in interaction with extracellular proteins (Callebaut
et al., BBRC 300: 619; 2003), two EGF-like domains and a
carboxy-terminal region rich in leucine and valine (Soncin et
VE-Statin/egfl7 Functions
during Physiological and
Tumoral Vascular
Development
Fabrice SONCIN
DR2 - Inserm
Group Members :
Lelièvre Etienne, CR Inserm
Mattot Virginie, CR CNRS
Pinte Sebastien, Postdoctoral fellow Inserm
Marchand Nathalie, Technician CNRS
Samson Chantal, Technician IPL
Villain Gaëlle, Technician CNRS
Delfortrie Suzanne, PhD Student CNRS/région
Poissonnier Loic, Student Master Lille 1
Lauridant-Philippin Géraldine, Student Master 2 Lille 2
Angiogenesis is the process by which new blood vessels arise
from the established vascular network in response to various
angiogenic stimuli. Angiogenesis is necessary to the formation
of organs during normal development and for the sustained
growth of most solid tumors (Carmeliet, Nat Med 6: 389; 2000).
Recently, several therapies which associated anti-angiogenic
compounds with chemotherapies proved to be effective for
treating colon (Hurwitz et al., N Engl J Med 350: 2335; 2004),
lung (Johnson et al., J Clin Oncol 22: 2184; 2004) and kidney
(Escudier et al., N Engl J Med 356 : 125 ; 2007) cancer patients,
clearly supporting the idea that inhibition or modulation of
angiogenesis is pertinent for treating cancer.
Initially, budding capillaries are mainly composed of
endothelial cells which form a monolayer at the inner border
of all blood vessels, in direct contact with the blood
circulation. Several proteins are specifically or quasiexclusively expressed by endothelial cells and define the
endothelial identity. They include the vascular endothelial
growth factor (VEGF) receptors Flt-1 (Shibuya et al., Oncogene
5 : 519; 1990 - de Vries et al., Science 255 : 989 ; 1992) and Flk1 (Terman et al., BBRC 187 : 1579 ; 1992 - Yamaguchi et al.,
Development 118 : 489 ; 1993), the Tie receptors (Partanen et
al., Mol Cell Biol 12 : 1698 ; 1992 - Sato et al., Proc Natl Acad Sci
USA 90: 9355; 1993 - Schnürch & Risau, Development 119: 957;
1993) or the adherens junction molecule VE-cadherin
(Lampugnani et al., J Cell Biol 118 : 1511 ; 1992 - Breier et al.,
Blood 87: 630; 1993). These markers are involved in essential
endothelial functions such as proliferation, survival and the
maintenance of the endothelium integrity. During
angiogenesis, they participate to the stabilization and
maturation of the initial capillaries into fully functional vessels
which also depend on the recruitment and interactions of the
endothelial cells with mural cells, i.e. smooth muscle cells
(SMC) and pericytes (Carmeliet, Nat Med 6: 389; 2000). The
complex molecular dialog that takes place during this phase
is decisive for the maintenance or the trimming of the newly
established capillaries and several molecules that are
produced by either or both cell types and that participate to
these interactions have been identified : endothelial cells
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Cancer
al., Embo J 22: 5700; 2003 - Parker et al., Nature 428: 754; 2004).
We previously demonstrated that VE-statin/egfl7 is expressed
in the endoplasmic reticulum and secreted in the cell
supernatant. VE-statin/egfl7 is an inhibitor of smooth muscle
cell migration in vitro and the first factor known to be
specifically expressed by endothelial cells which represses
SMC migration. (Soncin et al., Embo J 22: 5700; 2003). Using
the zebrafish knock-down model, Parker and colleagues
showed that VE-statin/egfl7 may be involved in vascular
tubulogenesis (Parker et al., Nature 428: 754; 2004). In fishes
where VE-statin/egfl7 is repressed, the two primary large
vessels are formed but tubulogenesis is impaired and
secondary vessels do not form 30 hr post-hatching although
their initial budding takes place (Parker et al., Nature 428 : 754 ;
2004). The molecular mechanisms that are responsible for
these defects are not known. More recently, a first approach
of inactivation of the gene in the mouse showed several
interesting points: the absence of expression of VE-statin/egfl7
induces an embryonic mortality of approximately 50%, the
aborted embryos present important edemas whose causes
were not analyzed. Surviving mice have many vascular defects
and hypoxic areas in several organs (heart, retina, CNS) as well
as tortuous vessels, of irregular diameter, or associated in
layers. The formation of the initiating capillaries of
angiogenesis is disturbed in these animals where the number
of capillaries in formation is increased, the `tips cells' and
`stalks cells' are more abundant and organized in multiple
cellular layers, which leads to a disturbed formation of these
capillaries. The protein seems to associate with the
extracellular matrix and to take part in the migration of the
endothelial cells (Schmidt et al., Development 134: 2913;
2007).
The few articles that have been published so far on
VE-statin/egfl7 confirmed our initial observations on the
specific expression of the gene in endothelial cells and on a
role of protein in the formation of blood-vessels.
C. Endothelial specific-transcriptional control of VE-statin/egfl7 gene
The VE-statin/egfl7 gene is specifically expressed in endothelial
cells and we have studied the regulatory processes that
govern the expression of the gene in these cells. The
expression is regulated by epigenetic processes which involve
the acetylation of histones H3 and H4 along the two
promoters. An enhancer region located 8,5kb ahead of the
initiation site allows a strong increase in activity in endothelial
cells compared to shorter fragments. An exhaustive analysis
of this enhancer has shown that it is important as a whole for
expression in endothelial cells. In the more proximal region of
the initiation sites, a region contains most of the endothelial
specificity and we have identified two ETS-binding sites that
are essential for expression. The 8,5 kb region contains most
of the information that is necessary for the endothelial
expression of the LacZ gene in vivo.
II. Role of VE-statin/egfl7
Role of VE-statin/egfl7 in vivo in blood vessel elastin
deposition
We recently generated transgenic mice where VE-statin/egfl7
expression was placed under the control of the keratin-14
promoter (K14-VE-statin/egfl7 mice). The HA-tagged VEstatin/egfl7 protein is thus ectopically produced by
keratinocytes. Transgenic pups derived from two independent
founders showed macroscopic defects characterized by a skin
flaky appearance and a necrotic tail tip. Transgenic pups
suffering the most severe phenotype usually die within 7 days
after birth. Immunohistochemistry of tail sections reveal that
VE-statin/egfl7 is expressed in keratinocytes, as expected.
Surprisingly, the VE-statin/egfl7 protein also accumulates
around the vascular bundles, more specifically around the main
tail artery. This indicated that VE-statin/egfl7 has an affinity for
vascular wall components. Ultra-structural analysis showed that
the elastin fibers are not correctly organized in the main tail
arteries of these transgenic animals. Further, an abnormal
endothelial and smooth muscle cell proliferation has also been
observed in the tail blood vessels. Such defects are similar to
those described in elastin-deficient mice (Li et al., Nature 393:
276; 2007), strongly suggesting a role for VE-statin/egfl7 in
elastogenesis. In vitro experiments demonstrated that
fibroblasts which normally deposit abundant amount of elastic
fibers, do not produce an elastin network when treated with VEstatin/egfl7. However, we showed that VE-statin/egfl7 does not
have an elastase activity, as it does not alter an already
deposited elastin network in vitro. Rather, VE-statin/egfl7
disturbs the elastin cross-linking process. We established the
underlying molecular mechanisms by demonstrating that the
VE-statin/egfl7 interacts with lysyl oxidases and inhibit their
enzymatic activity. We also demonstrated that endothelial cells
which are normally not able to deposit detectable amounts of
elastin, can be induced to do so after targeting the endogenous
VE-statin/egfl7 gene by RNA interference, thus showing that in
endothelial cells, VE-statin/egfl7 is a key inhibitor of
elastogenesis (Lelièvre et al., Embo J 27: 1658 ; 2008).
Recent studies performed by the team
I. Study of VE-statin/egfl7 gene expression and regulation
A. Analysis of expression in postnatal and adult normal tissues
The expression of VE-statin/egfl7 was studied in human and
mouse adult tissues by in situ hybridization. In all tissues
analyzed, VE-statin/egfl7 is weakly detected in a subset of
endothelial cells of the adult vasculature indicating that VEstatin/egfl7 is not required for the function of mature blood
vessels. VE-statin/egfl7 expression was also characterized
during the development of the mouse retina vasculature, a
well-described model of vascular growth, remodeling and
maturation.
B. Analysis of VE-statin/egfl7 expression in tumors
A preliminary study performed on a slide tissue-array revealed
that VE-statin/egfl7 expression is mainly detected in squamous
cell carcinomas in lung and uterus. Further analysis were
performed on human lung tissue-array slides and indicated
that VE-statin/egfl7 expression is up-regulated in T2 stage
compared to earlier or later stages suggesting that VEstatin/egfl7 may be at least a diagnostic marker for lung
cancer.
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Cancer
Publications
Cancer Biology
and Chemistry (CBC) :
2007
from Fundamental Research
to Therapeutic Applications
Oleg MELNYK
Le Bras A, Lionneton F, Mattot V, Lelièvre E, Caetano B, Spruyt N,
and Soncin F.
HIF-2α specifically activates the VE-cadherin promoter independently of
hypoxia and in synergy with Ets-1 through two essential ETS-binding sites.
Oncogene, 2007, 26:7480-7489.
DR2 - CNRS
2008
Group Members :
Gérard AC, Poncin S, Caetano B, Sonveaux P, Audinot JN, Feron O,
Colin IM, and Soncin F.
Iodine deficiency induces a thyroid stimulating hormone-independent
early phase of microvascular reshaping in the thyroid.
Am J Pathol, 2008, 172(3):748-760.
Chemistry group
Melnyk Oleg, DR2 CNRS
Ollivier Nathalie, Engineer Research IPL
Dheur Julien, Postdoctoral fellow CNRS
Ebran Jean Philippe, Postdoctoral fellow CNRS
Mhidia Reda, Postdoctoral fellow CNRS
Blanpain Annick, Technician IPL
Desmet rémi, Technician IPL
Drobecq Hervé, Technician Lille 2
Boll Emmanuelle, Technician CNRS
Besret Soizic, PhD Student MENRT
Lelièvre E, Hinek A, Lupu F, Buquet C, Soncin F, and Mattot V.
VE-statin/egfl7 regulate vascular elastogenesis by interacting with lysyl
oxidases.
Embo J, 2008, 27 : 1658-1670.
2009
Le Bras A, Soncin F. 2009
Genes that make the endothelial identity.
J Soc Biol, 2009, 203:125-141.
HGF-SF MET signaling in cancer
Fafeur Véronique, CR Inserm
Vicogne Jérôme, CR2 CNRS
Goormachtigh Gautier, Postdoctoral fellow CNRS
Mougel Alexandra, Engineer CNRS
Dewitte Caroline, ITA CNRS
Legoff Arnaud, PhD Student MENRT
Héliot L, Mattot V, Soncin F, Gougeon ML, Dombrowicz D, Capron M.
A functional gammadeltaTCR/CD3 complex distinct from gammadeltaT
cells is expressed by human eosinophils.
PLoS One, 2009, 4:e5926.
LMPI in cancer - Epigenetics and cancer
Adrienssens Eric, PR2 Univ Lille 1
Ndour Alioune Papa, ATER Lille 1
Berteaux Nathalie, postdoctoral fellow CNRS
Spruyt Nathalie, Engineer CNRS
Brocqueville Guillaume, PhD Student MENRT
Leclercq Berenice, Master 2
Sahin A, Vercamer C, Kaminski A, Fuchs T, Florin A, Hahne JC,
Mattot V, Pourtier-Manzanedo A, Pietsch T, Fafeur V, Wernert N.
Dominant-negative inhibition of Ets 1 suppresses tumor growth, invasion
and migration in rat C6 glioma cells and reveals differentially expressed
Ets 1 target genes.
Int J Oncol, 2009, 34:377-389.
2010
The CBC group results from the association between two
biologist groups (V. Fafeur and E. Adriaenssens) and one
chemist group stemming from UMR 8161. This association is
based on common previous collaborations, which already led
to publications and funded projects and to an expected
synergy between distinct and complementary expertises.
The heart of the CBC research project is an interdisciplinary
project focused on the discovery of novel therapeutic
strategies based on the extracellular inhibition of HGF/MET
signaling. The groups are of course also involved in their own
fundamental research project which are in support of the
common project: V. Fafeur explores the signaling of HGF/MET
in cancer, E. Adriaenssens studies the role of H19-related noncoding RNAs in the epigenetic control of tumor progression,
whereas O. Melnyk is involved in the design of novel peptide
ligation chemistries and micro-nanotechnologies which are
combined for studying some biomolecular interactions.
O. Melnyk is responsible for the Peptide Chemistry Systems
Biology (Peptide CSB) platform, a technological platform
(http://csb.ibl.fr). This platform accommodates a start-up
company founded by V. Souplet, C. Ollivier and O. Melnyk
(created July 2008, http://www.innobiochips.fr/).
Le Bras A, Samson C, Trentini M, Caetano B, Lelievre E, Mattot V,
Beermann F, Soncin F.
VE-statin/egfl7 expression in endothelial cells is regulated by a distal
enhancer and a proximal promoter under the direct control of erg and
GATA-2.
PLoS One, 2010, 5(8).
PhD
Alexandra LE BRAS
Directeur : F. Soncin
Decembre 2007
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Cancer
have described for the first time the synthesis of peptide
thioesters using the Fmoc strategy and a N,S-acyl shift on the
solid phase (Ollivier et al., Org Lett 7: 2647; 2005).
Now our main activity is focussed on the design of novel
native ligation chemistries for the total synthesis of proteins
of moderate size (<200 amino acids).
1-Chemistry group (O. Melnyk)
The design of novel ligation chemistries leading to the
formation of native or non native peptide bonds at the ligation
site represents one of the greatest challenges in the peptide
field. This research field includes the development of solid
phase synthetic strategies allowing an easy access to properly
functionalized peptides ready for ligation. The ligation
chemistries are also used for linking biomolecules to surfaces
of different kind (silicon oxide planar substrates, silica or gold
nanoparticles, silicon nanowires…) within miniaturized
devices for studying the binding properties of biomolecules,
for characterizing the chemical properties of nanodevices, or
for the controlled assembly of inorganic materials.
Peptides and nanodevices (collaborations : D. Stiévenard,
IEMN, Lille)
The group has collaborated also with researchers from IEMN
(D. Stiévenard) and IRI (R. Boukherroub) for several years.
Recent work concerns the development of novel biosensors
which allow the electrical detection of biomolecular
interactions. The principle of the detection is based on the
induction of a tunnel effect between nanoelectrodes
separated by a few tens of nm, following the specific insertion
of gold nanoparticle-labelled reagents into the nanogap. The
use of biological samples (serum) demonstrated the
usefulness of the biosensor in a real detection experiment
(detection of antibodies by arrayed antigens). This work was
published in several papers recently (Haguet et al., Applied
Phys Lett 84: 1213; 2004 – Brinkmann et al., Langmuir 23: 9784;
2006 – Marcon et al., Biosensors Bioelectronics 23: 81; 2007 –
Marcon et al., Biosensors Bioelectronics 23: 1185; 2008 – ElMahdi et al., chapter book In: Colloidal Nanoparticles in
Biotechnologies, 2008 – Marcon et al., Bioconjugate Chem 802;
2008). Now our work is focused on the electrical detection of
biomolecular interactions using silicon nanowires. We have
recently shown that protocollagen molecules can be used for
controlling the positioning of silicon nanowires on gold
microelectrodes. This self-assembly method allows the
speeding up of the fabrication of silicon nanowire based chips,
which will be further used for studying heparin sulphate
cleavage by heparanase, an enzyme involved in metastasis
and angiogenesis.
Peptide ligation chemistry in solution and on surfaces
(collaborations : J.-O. Durand, UMR 5253, Montpellier; D.
Stiévenard, IEMN, Lille; R. Boukherroub, IRI, Lille)
Since 2004, we have developed efficient methods for
synthesizing and characterizing glyoxylyl peptides, which are
useful peptide derivatives in ligation chemistry (Far et al.,
Tetrahedron Lett 45: 1271; 2004 – Melnyk, Tetrahedron Lett 45:
7163; 2004 – Far et al., J Peptide Sci 11: 424; 2005 – Far et al.,
Tetrahedron 61: 6138; 2005- Garcia et al., J Peptide Sci 12: 734;
2006), and we have deeply explored the interest of
semicarbazide/glyoxylyl peptide ligation for the synthesis of
biologically active compounds (Far et al., Bioorg Med Chem
Lett 14: 4439; 2004 – Bourel-Bonnet et al., Bioconjug Chem 16:
450; 2005 – Dubs et al., J Comb Chem 9: 873; 2007), for the
preparation of hybrid bioorganic-inorganic materials (Joly et
al., Eur J Org Chem 2473; 2005 – Doussineau et al., Eur J Inorg
Chem 2766; 2006) and for the fabrication of oligonucleotide
or peptide microarrays (Duburcq et al., Bioconjugate Chem 15:
307; 2004 – Duburcq et al., Bioconjugate Chem 15: 317; 2004
- Hot et al., Curr Prot Nucleic Acid Chem 12.6 : 1; 2004 –
Cofinier et al., Langmuir 21: 1489; 2005). Peptides are also
useful tools for characterizing the chemical properties of
surfaces. For example, cobalt-carbonyl peptide complexes
were used successfully for characterizing the site-specific
ligation of peptides to surfaces by FTIR spectroscopy (Olivier
et al., Langmuir 22: 7059; 2006).
Silicon oxide is a useful substrate for making microarrays, and
we have like others developed chemical methods for
attaching molecules to such surfaces, especially microscope
glass slides. The functionalization of plastic substrates is still
in its infancy and their use for the site-specific immobilization
of peptides is challenging. Recent results from the lab concern
the synthesis of semicarbazide modified silica nanoparticles
which were used for the chemical micropatterning of
polycarbonate (PC) (Souplet et al., Chem Bio Chem 8: 315;
2007 – Souplet et al., Chapter book in Peptide Microarrays,
Humana Press, 2008), an organic polymer often used for the
preparation of microfluidic devices. Glyoxylyl-peptides were
site-specifically linked to PC through the silica nanoparticle
layer and used for capturing antibodies. An interesting
curvature effect was observed by microarraying a set of
nanoparticles with a diameter ranging from 30 to 300 nm: the
best specificity of interaction was observed for the smallest
particles.
Besides glyoxylyl peptide chemistry, the group has been
involved since 2005 in the development of novel solid phase
methods for synthesizing peptide thioesters, which are
important peptide derivatives in native ligation chemistry. We
Peptide microarrays, post-translational modifications and
chips from chips (collaboration : C. Locht, IBL, Lille)
The group develops novel microarray methods in the peptide
microarray field (see the first paragraph). We have prepared
complex peptide microarrays presenting unmodified peptides
and methylated peptides for studying the humoral response
in mice and humans produced after immunizing with HBHA
(heparin-binding haemagglutinin present on the surface of
M. Tuberculosis), or infection by M. Tuberculosis. The C-terminal
domain of HBHA presents a complex methylation pattern and
the peptide microarray technology is a powerful tool for
studying such a complex protein. This project allowed us to
explore the “chips from chips” concept : a peptide microarray
obtained by microarraying unmodified peptides can be
treated chemically to introduce specifically post-translational
modifications on given amino acids. In the present case, the
microarray was methylated on lysines. We could demonstrate
that the information provided by the in situ methylation
method is similar to the information provided by the
microarrayed methylated peptides. Overall, we believe that
the method will speed up the study of some post-translational
modifications and we will develop in the future methods
allowing the control of the modification pattern within a given
peptide on the microarray (collaboration with R. Boukherroub,
IRI, Lille).
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Cancer
capable itself to induce apoptosis. As a functional
consequence, the cleaved MET receptor contributes to the
amplification of apoptosis (Tulasne et al., Mol Cell Biol 24:
10328; 2004 - Foveau et al., Cell Death Diff 14: 752; 2007 Deheuninck et al., BBRC, 367: 573; 2008). In agreement with
the survival function of HGF/SF, we also demonstrated that
HGF/SF unfavors the caspase cleavage of MET. The mechanism
involves the phosphorylation of MET and the recruitment of
the CBL protein ligase to a phosphorylated tyrosine adjacent
to the juxtamembrane caspase cleavage of MET, which may
prevent the accessibility of this MET region to caspases. Taken
together, these results open the possibility that MET is a
dependence receptor, such a receptor induces survival in the
presence of its ligand and induces apoptosis in its absence
(Bredesen et al., Cell Death Differ 12 : 1031; 2005). It should be
stressed that various fragments and truncated versions of MET
have been detected in many cancer cells, nonetheless their
mode of generation and functions have never been described
before.
2-Biology group - HGF/SF-MET
signaling in cancer (V. Fafeur)
Hepatocyte growth factor/scatter factor (HGF/SF) is the ligand
of the MET tyrosine kinase receptor, initially discovered as an
oncogene (Birchmeier et al., Nat Rev Mol Cell Biol 4: 915; 2003
- Corso et al., Trends Mol Med 11: 284; 2005). HGF/SF is a
disulfide-linked heterodimer consisting of a 62-kDa subunit
and a 34 kDa subunit, and the MET receptor is a disulfidelinked heterodimer consisting in an extra-cellular 50 kDa
chain and a 140 kDa chain spanning the membrane and
containing the catalytic domain. The structure of HGF/SF and
MET, including their interactions with heparin, are constantly
improved (Kemp et al., Biochem Soc Trans 34 : 414; 2006 Holmes et al., J Mol Biol 367 : 395; 2007).
The ligand-activated-MET stimulates survival and proliferation
of many cell types, and characteristically, stimulates the
scattering and morphogenesis of epithelial cells. The genetic
analysis of HGF/SF and MET in mice has shown their essential
roles during the development of the placenta, liver, muscles
and neurons (Birchmeier & Gherardi, Trends Cell Biol 8 : 404;
1998). The therapeutic applications of HGF/SF in liver and
renal disease and tissue regeneration are currently explored
(Matsumoto & Nakamura, BBRC 239 : 639; 1997 - Zou et al.,
Cancer Res 67 : 4408; 2007).
The deregulation of the HGF/SF-MET signaling system occurs
in human tumors via multiple mechanisms, including
coexpression of HGF/SF and MET, receptor amplification and
point mutations, and is often associated with metastatic
progression (reviewed in (Birchmeier et al., Nat Rev Mol Cell
Biol 4 : 915; 2003 - Corso et al., Trends Mol Med 11: 284; 2005).
HGF/SF and MET are overexpressed in many tumors
(carcinomas, sarcomas, blood malignancies...), often
correlating with poor prognosis. Direct evidence that
implicates MET in human cancers is provided by activating
mutations discovered in both inherited and sporadic forms of
human renal papillary carcinoma (Schmidt et al., Nat Genet
16 : 68; 1997). Such mutations have also been found in other
types of cancer and metastasis. HGF/SF and MET are therefore
attractive candidates for targeted cancer therapy.
A molecular understanding of how this receptor is switched
on and off will provide the basis for developing rational
interventions in tumor therapy.
The GAB1 protein is a major partner of the MET receptor. This
multiadaptator binds directly to the C-terminal part of the
MET receptor, whereas it binds indirectly to other membrane
bound receptors. Strong similarities were found between the
phenotypes of mouse embryos GAB1-/- et MET-/- and cells
stemmed from these embryos allowed to demonstrate the
specific contribution of GAB1 in signal transduction by HGF/SF
(Sachs et al., J Cell Biol 150: 1375; 2000). While GAB1 is known
to amplify and diversify signaling by MET, its possible role in
the negative regulation of MET is not known. This led us to
examine the fate of GAB1 upon apoptotic stress induction,
similarly to the above described fate of MET. We have shown
that GAB1 is also cleaved by caspases, whereas ERK, another
intracellular target of MET, was not modified. Several sites of
caspase cleavages were identified in GAB1 and they generate
several fragments, including a fragment containing the METbinding domain and a fragment corresponding to the
C-terminal part of GAB1, with essential recruitment sites for
the SHP2 phosphatase. These fragments may act in a
dominant negative manner on MET and GAB1 signaling. In
parallel, we also investigated the possible down regulation of
GAB1 in response to HGF/SF. The MET receptor is known to be
degraded and recycled by endocytosis by mechanisms
implicating its ubiquitination, following its activation by
HGF/SF (Peschard et al., Mol Cell 8: 995; 2001 - Petrelli et al.,
Nature 416: 133; 2002). We found that GAB1 is also modified
by ubiquitination following treatment of cells by HGF/SF and
the mechanism involves the recruitment of CBL, an ubiquitin
ligase, which is also known to favor ubiquitination of MET.
Taken together, our data revealed an original pro-apoptotic
function of MET in response to stress, which contrasts with its
known survival effects in response to HGF/SF. Mechanistic
studies have been undertaken to understand how MET can
have opposite functions in distinct physiological environment,
including by exploring the fate of GAB1, another essential
component of the MET signaling complex. This fundamental
research is our basic expertise and feeds the project with the
chemists consisting in identifying MET inhibitors for anticancer strategies.
The group has been investigating signal transduction by
HGF/SF-MET, leading to cell scattering, morphogenesis and
survival, for more than 10 years. In particular, this led us to
describe the contribution of the RAS pathway in multiple
biological responses to HGF/SF, implicating the ERK and JNK
MAP kinases and the ETS1 transcription factor (Fafeur et al.,
Cell Growth Differ 8: 655; 1997 - Tulasne et al., Mol Biol Cell 10:
551; 1999 - Paumelle et al., Mol Biol Cell 11: 3751 ; 2000 Paumelle et al., Oncogene 21: 2309; 2002). We also identified
novel target genes of HGF/SF, implicated in its survival effects
(Leroy et al., Annal NY Acad Sci 1090: 188; 2006). More recently,
we found an original pro-apoptotic function of MET, which is
evidenced in the absence of HGF/SF. Upon apoptotic stress, a
double and sequential caspase cleavage within the MET
intracellular part (juxtamembrane and C-terminal) generates
an extracellular fragment (p100 MET) still bound to the
membrane and acting as a decoy receptor, and an intracellular
fragment (p40 MET), containing the kinase domain and
These past years, this project has been granted by the
“Fondation de France”, ARC and the “Ligue Régionale Contre
le Cancer-Comité Nord”. The results allowed J. Deheuninck and
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B. Foveau to receive their PhD from the University of Lille 1 in
November 2006 and November 2007, respectively.
David Tulasne decided to create his own group within the
laboratory, which led him to obtain an ANR grant “Jeunes
Chercheurs” in 2007 and to start an autonomous group in
January 2008. According to our common history, our two
groups are still investigating MET signaling in cancer, with
specific fields of research being explored. Notably, my group
is now starting a project for identifying extracellular MET
inhibitors. We can anticipate that the understanding of their
mode of action will benefit of the concept that MET can be a
proapoptotic actor. Based on ongoing interactions,
cooperation between the two groups is therefore most likely.
LCLs as well as TE1 and NC5 cell lines are LMP1-dependent.
Our goal was then to use LMP1-derived molecules and/or
other molecules selectively inhibiting part of the LMP1triggered signaling pathways to challenge them in these
different EBV-infected cellular models, as follows : (i)
Phenotype outputs associated with selective recruitment of
various proximal molecules to LMP1, (ii) the relative
importance of ensuing signal transduction pathways
emerging from these interactions in the EBV-dependent
proliferation and transformation processes. This approach is
developed using inducible vectors expressing different parts
of LMP1, cell-penetrating peptides derived from the LMP1
regions interacting with adaptors (in collaboration with Oleg
Melnyk group) and small double strand RNAs (siRNAs for small
inhibitory RNAs) raised against some LMP1 adaptors and
signalling. These approaches allowed us to demonstrate that
LMP1 activates its own promoter (pLMP1) through the JNK
signaling pathway. Our results also reveal that this activation
is tightly controlled by LMP1, since pLMP1 is inhibited by
LMP1-activated NF-B signaling pathway. By using our
physiological models of EBV-infected cells displaying type II
latency as well as LCL expressing a type III latency, we also
demonstrate that this balanced autoregulation of LMP1 is
shared by both latency programs. Finally, we show that this
autoactivation is the most important mechanism to maintain
LMP1 expression during the type II latency program of EBV
(Goormachtigh et al., J Virol 80: 7382; 2006).
Using our dominant negative forms of LMP1, and in
collaboration with Jean Feuillard’s laboratory (Limoges), we
have demonstrated that LMP1 is able to sensitize B cells to
induction of CD95-mediated apoptosis (Le Clorennec et al.,
Blood 107: 2070; 2006). LMP1 appears to have opposite effects
in latency III program: survival versus apoptosis. Molecular basis
of this apoptosis has been studying more extensively (Le
Clorennec et al., J Virol 82: 6721; 2008). In addition, we used the
dominant negative forms of LMP1 to inhibit the TNF signalling
and the results improved the knowledge on TNFR biology (In
preparation). In our cells (latency II program), we have shown
that LMP1 is also able to induce apoptosis (In preparation).
These opposite effects of LMP1 are similar to those observed
for MET and more generally for numerous oncogenes, which
can share survival / apoptotic properties. This apoptotic effect
of LMP1 is explored in collaboration with V Fafeur’s group.
3-Biology group (E. Adriaenssens)
-LMP1 signaling in cancer
Epstein–Barr virus (EBV), the etiological agent of infectious
mononucleosis, is a human herpesvirus involved in the
development of several human malignancies. EBV expression
is tightly associated with the transformed status of infected B
lymphocytes and epithelial cells in B lymphomas and
carcinomas, and is also observed in some T lymphomas.
Recently, the involvement of EBV in gastric and mammary
carcinomas has been assumed. In transformed cells, EBV is
found in a latent state and only few viral genes are expressed.
Eleven genes are expressed in LCLs displaying a type III
latency, whereas a type II latency restricted to expression of
three genes is encountered in EBV-transformed T cells and
monocytes. This last transcription pattern is typically found in
most of the EBV-associated malignancies.
The product of one of these genes is latent membrane
protein-1 or LMP1 (Kaye et al., Proc Natl Acad Sci USA 90: 9150;
1993). LMP1 is an integral membrane molecule (a 63 kDa
plasma membrane protein with six transmembrane
segments) expressed by EBV during viral latency and displays
properties of a constitutively activated member of the TNF
receptor family (Eliopoulos & Young, Semin Cancer Biol 11:
435; 2001). Spontaneous oligomerization mediated by
transmembrane and cytoplasmic N-terminal fragments
induces activation of several signaling pathways. LMP1 is
required for B-cell or monocyte immortalization induced by
EBV and is sufficient to transform rodent fibroblasts.
Transforming potential of LMP1 is mediated by its cytoplasmic
C-terminal domain, which activates various cellular signaling
pathways including NFB (Eliopoulos et al., Oncogene 14: 2899;
1997) and JNK (Eliopoulos et al., J Virol 73: 1023; 1999).
Our specificity in the field of EBV research is to have isolated
specific cellular models to study LMP1 signaling (Groux et al.,
Blood 89: 4521; 1997 - Masy et al., J Virol 76: 6460; 2002), as
well as to develop specific tools conceptually exploiting LMP1derived Dominant Negative (DN) properties (Adriaenssens et
al., Oncogene 23: 2681; 2004). We discovered EBV-infected
monocytic (TE1) and T lymphoid (NC5) cells, infected and
immortalized by EBV, displaying type II latency. Indeed,
whereas the majority of malignancies due to EBV display type
II latency, the only cellular model available is LCL displaying a
type III latency. These cell lines were obtained after in vitro
infection and transformation of human peripheral blood cells.
They both express a type II viral latency (EBNA1, LMP1 and
LMP2) characteristic of a viral expression pattern found in the
majority of EBV-infected tumors arising in vivo from
immunocompetent patients. The survival and proliferation of
Due to the recent decease of Jean Coll, the group leader, and
thanks to thematic convergences with Véronique Fafeur, we
have decided to join her to study MET signaling.
Non coding H19-RNA related genes, epigenetics and cancer
For several years, we have been working on the non-coding H19
gene, in particular in breast cancer progression. We have
demonstrated its implication in mammary, endometrial and
prostatic tumorigenesis (Adriaenssens et al., Am J Pathol 153:
1597; 1998 - Berteaux et al., J Endocrinol 183: 69; 2004 - Lottin
et al., Eur J Cancer 41: 168; 2005). We have shown that H19
enhances the tumorigenic properties of breast cancer cell lines
(Lottin et al., Carcinogenesis 23: 1885; 2002) and promotes the
G1/S transition (Berteaux et al., J Biol Chem 280: 29625; 2005).
Recently, we have discovered 91H, at the H19 locus, as a new
transcript implicated in H19 and IGF2 (neighbouring gene)
transcription, as well as genomic imprinting. This emerging
project will be continued.
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Cancer
Marcon L, Stiévenard D, Melnyk O.
Electrical detection of antibodies from human serum based on the
insertion of gold-labeled secondary antibodies into micro or nanogaps.
In : Colloidal Nanoparticles in Biotechnologies. Elaissari, A. Ed.
Published by John Wiley & Sons, US, 2008, in press.
Publications
Chemistry group
2007
Characterization of Nanogap Chemical Reactivity Using Peptide-Capped
Gold Nanoparticles and Electrical Detection.
Bioconjugate Chem, 2008, 19:802-805.
Coffinier, Y., S. Szunerits, C. Jama, R. Desmet, O. Melnyk, B. Marcus,
L. Gengembre, E. Payen, D. Delabouglise and R. Boukherroub. 2007
Peptide immobilization on amine-terminated boron-doped diamond surfaces.
Langmuir 23, 4494-4497.
Pamelard F. Even G, Apostol C, Preda C, Dhaenens C, Desmet R,
Melnyk O.
PASE : a web-based platform for peptide/protein microarray experiments"
Peptide Microarrays, Humana Press, Marina Cretich and Marcella Chiari
Eds., chapter book, 2008, in press.
Coffinier Y, Szunerits S, Marcus B, Desmet R, Melnyk O, Gengembre L,
Payen E, Delabouglise D,Boukherroub R.
Covalent linking of peptides onto oxygen-terminated boron-doped
diamond surfaces.
Diamond Related Materials, 2007, 16:892-898.
Piret G, Coffinier Y, Roux C, Melnyk O, Boukherroub R.
Biomolecule and Nanoparticle Transfer on Patterned and Heterogeneously
Wetted Superhydrophobic Silicon Nanowire Surfaces.
Langmuir, 2008, 24:1670-1672.
Dubs P, Bourel-Bonnet L Subra G, Blanpain A, Melnyk O, Pinel AM,
Gras-Masse H, Martinez J.
Parallel synthesis of a lipopeptide library by hydrazone-based chemical ligation.
J Comb Chem, 2007, 9:873-881.
Souplet V. Roux C, Melnyk O.
Peptide microarrays on bisphenol A polycarbonate.
Peptide Microarrays, Humana Press, 2008, Marina Cretich and Marcella
Chiari Eds., chapter book, 2008, in press.
Electrical detection of human immunoglobulins G from human serum
using a microbiosensor.
Biosensors Bioelectronics, 2007, 23:81-87.
2009
Melnyk O, I. G. Stara IG, I. Stary I, B. Klepetarova, and D. Saman. 2007
Reaction of isocyanate-functionalised silicon wafers with complex amino
compounds.
Eur J Org Chem 4032-4037.
Baud C, Hodak H, Willery E, Drobecq H, Locht C, Jamin M, JacobDubuisson F.
Mol Microbiol, 2009, 74:315-329.
Roux C, Chai F, Ollivier N, Winter S, Melnyk O, Hildebrandt HF.
Ti-Cp functionalization by deposition of organic/inorganic silica nanoparticles.
Biomol Engineer, 2007, 24:549-554.
Fertier L, Cretin M, Rolland M, Durand JO, Raehm L, Desmet R,
Melnyk O, Zimmermann C, Dejous C, Rebiere D.
Love wave immunosensor for antibody recognition using an innovative
semicarbazide surface functionalization.
Sensors Actuators B-Chem, 2009, 140:616-622.
Souplet V, Carion O, Maillet C, Olivier C, Médard N, Durand JO,
Melnyk O.
Chemicals micropatterning of polycarbonate for site-specific peptide
immobilization and biomolecular interactions.
ChemBioChem, 2007, 8:315-322.
Helleboid-Chapman A, Nowak M, Helleboid S, Moitrot E, Rommens C,
Dehondt H, Heliot L, Drobecq H, Fruchart-Najib J, Fruchart JC.
Apolipoprotein A-V modulates insulin secretion in pancreatic beta-cells
through its interaction with midkine.
Cell Physiol Biochem, 2009, 24:451-460.
Souplet V, Desmet R, Melnyk O.
Imaging of protein layers with an optical microscope for the
characterization of peptide microarrays.
J Pept Sci, 2007, 451-457.
Ollivier N, Besret S, Blanpain A, Melnyk O.
Silver-Catalyzed azaGly Ligation. Application to the Synthesis of
Azapeptides and of Lipid-Peptide Conjugates.
Bioconjug Chem, 2009, 20:1397-1403.
2008
Allan G, Barbet S, Coffinier Y, Delerue C, Deresmes D, Diarra M,
Diesinger H, Grandidier B, Marcon L, Mélin T, Melnyk O,
Stiévenard D, Wirtz L, Zdrojek M.
Fundamental studies in nanosciences at the institute of electronics,
microelectronics, and nanotechnology (IEMN).
Int J Nanotechnol, 2008, 5:631-649.
Pamelard F, Even G, Apostol C, Preda C, Dhaenens C, Fafeur V,
Desmet R, Melnyk O.
PASE: A Web-Based Platform for Peptide/Protein Microarray Experiments.
Methods Mol Biol, 2009, 570:413-430.
Salhi B, Vaurette F, Grandidier B, Stievenard D, Melnyk O,
Coffinier Y, Boukherroub R.
The collagen assisted self-assembly of silicon nanowires.
Nanotechnology, 2009, 20.
Besret S, Ollivier N, Blanpain A, Melnyk O.
Chemoselective peptide ligation using phenylthiocarbamate chemistry.
J Org Chem, 2008, accepted.
Stechly L, Morelle W, Dessein AF, Andre S, Grard G, Trinel D,
Dejonghe MJ, Leteurtre E, Drobecq H, Trugnan G, Gabius HJ, Huet G.
Galectin-4-regulated delivery of glycoproteins to the brush border
membrane of enterocyte-like cells.
Traffic, 2009, 10:438-450.
El-Mahdi O, Souplet V, Carion O, Roux C, Garcia JM, Maillet C,
Olivier C, Durand JO, Melnyk O.
Semicarbazide/ -oxo aldehyde site-specific ligation chemistry. From
peptide microarrays to the micropatterning of polycarbonate or titanium
oxide using silica nanoparticles.
In : Colloidal Nanoparticles in Biotechnologies. O. Melnyk corresponding
author. Elaissari, A. Ed. Published by John Wiley & Sons, 2008, US.
In Situ Ligation between Peptides and Silica Nanoparticles for Making
Peptide Microarrays on Polycarbonate.
Bioconjug Chem, 2009, 20:550-557.
Marcon L, Melnyk O, Stiévenard D.
Current based antibodies detection from human serum enhanced by secondary
antibodies labelled with gold nanoparticles immobilized in a nanogap.
Biosensors Bioelectronics, 2008, 23:1185-1188.
Souplet V, Roux C, Melnyk O.
Peptide microarrays on bisphenol a polycarbonate.
Methods Mol Biol, 2009, 570:287-297.
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Cancer
Lagadec C, Adriaenssens E, Toillon RA, Chopin V, Romon R,
Van Coppenolle F, Hondermarck H, Le Bourhis X.
Tamoxifen and TRAIL synergistically induce apoptosis in breast cancer cells.
Oncogene, 2008, 27:1472-1477.
HGF-SF MET signaling in cancer
2007
Ji Z, Degerny C, Vintonenko N, Deheuninck J, Foveau B, Leroy C,
Coll J, Tulasne D, Baert JL, Fafeur V.
ubiquitinylation.
Oncogene, 2007, 26: 395-406.
Le Clorennec C, Ouk TS, Youlyouz-Marfak I, Adriaenssens E, Coll J,
Feuillard J, Jayat-Vignoles C.
Molecular basis of Epstein-Barr Virus Latent Membrane Protein 1 (LMP1)
Cytotoxicity in EBV Latency III B-cells : LMP1 induces type II Ligand independent Autoactivation of CD95/FAS with Caspase-8 mediated Apoptosis.
J Virol, 2008, 82:6721-6733.
Foveau B, Leroy C, Ancot F, Deheuninck J, Ji Z, Fafeur V., Tulasne D.
Amplification of apoptosis through sequential caspase cleavage of the
MET tyrosine kinase receptor.
Cell Death Differ, 2007, 14:752-764.
2009
Lagadec C, Meignan S, Adriaenssens E, Foveau B, Vanhecke E,
Romon R, Toillon RA, Oxombre B, Hondermarck H, Le Bourhis X.
TrkA overexpression enhances growth and metastasis of breast cancer cells.
Oncogene, 2009, 28:1960-1970.
Yan Y, Tulasne D, Browaeys E, Cailliau K, Khayath N, Pierce RJ,
Trolet J, Fafeur V, Younes AB, Dissous C.
Molecular cloning and characterisation of SmSLK, a novel Ste20-like
kinase in Schistosoma mansoni.
Int J Parasitol, 2007, 37:1539-1550.
2010
Cazet A, Lefebvre J, Adriaenssens E, Julien S, Bobowski M,
Grigoriadis A, Tutt A, Tulasne D, Le Bourhis X, Delannoy P.
GD3 synthase expression enhances proliferation and tumor growth of
MDA-MB-231 breast cancer cells through c-Met activation.
Mol Cancer Res, 2010, Oct 1.
2008
Deheuninck J, Foveau B, Goormachtigh G, Leroy C, Ji Z, Tulasne D,
Fafeur V.
Caspase cleavage of the MET receptor generates an HGF interfering fragment.
Ndour PA, Ouk TS, Brocqueville G, Mougel A, Vanhecke E, Feuillard
J, Coll J, Adriaenssens E.
Inhibition of tumor necrosis factor-induced phenotpypes by short
intracellular versions of latent membrane protein-1.
Cell Signal, 2010, 22:303-313.
Hahne JC, Okuducu AF, Sahin A, Fafeur V, Kiriakidis S, Wernert N.
The transcription factor ETS-1 : its role in tumour development and
strategies for its inhibition.
Mini Rev Med Chem, 2008, 8:1095-1105. Review.
Romon R, Adriaenssens E, Lagadec C, Germain E, Hondermarck H,
Le Bourhis X.
Nerve growth factor promotes breast cancer angiogenesis by activating
multiple pathways.
Mol Cancer, 2010, 9:157.
Vicogne J, Pessin JE.
SNARE-mediated in vitro fusion assay.
Methods Mol Biol, 2008, 457:241-251.
2009
Verbeke S, Meignan S, Lagadec C, Germain E, Hondermarck H,
Adriaenssens E, Le Bourhis X.
Overexpression of p75(NTR) increases survival of breast cancer cells
through p21 (waf1).
Cell Signal, 2010, 22:1864-1873.
Villeret V, Tulasne D, Fafeur V.
Phosphorylation of the MET receptor in juxtamenbrane tyrosine residue
1001 inhibits its caspase-dependent cleavage.
Cell Signal, 2009, 211455-1463.
Foveau B, Ancot F, Leroy C, Petrelli A, Reiss K, Vingtdeux V, Giordano
S, Fafeur V, Tulasne D.
Down-regulation of the met receptor tyrosine kinase by presenilindependent regulated intramenbrane proteolysis.
Mol Biol Cell, 2009, 20:2495-507.
PhD
Chemistry group
Sahin A, Vercamer C, Kaminski A, Fuchs T, Florin A, Hahne JC,
Mattot V, Pourtier-Manzanedo A, Pietsch T, Fafeur V, Wernert N.
ans migration in rat C6 glioma cells and reveals differentially expressed
Ets 1 target genes.
Int J Oncol, 2009, 34:377-389.
Vianney SOUPLET
Directeurs : O. Melnyk & D. Ausséré
PhD « Chemistry »
Février 2008
2008
Gauthier GOORMACHTIGH
Directeur : J. Coll
20 juin 2006
Adriaenssens E, Vanhecke E, Saule P, Mougel A., Page A, Romon R,
Nurcombe V, Le Bourhis X and H. Hondermarck.
Nerve growth factor is a potential therapeutic target in breast cancer.
Cancer Res, 2008, 68:346-351
Berteaux N, Aptel N, Cathala G, Janton C, Coll J, Spruyt N,
Hondermarck H, Dugimont T, Curgy JJ, Forne T, Adriaenssens E.
A H19 antisense RNA overexpressed in breast cancer contributes to
paternal IGF2 expression.
Mol Cell Biol, 2008, 28:6731-6745.
Alioune NDOUR PAPA
Directeur : E Adriaenssens
3 décembre 2009
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Cancer
systematically in all cultures, whatever the donor, while most
senescent cells died. These post-senescent (PS) emerging cells
were found to have resumed normal PCNA, p16 and p21
expression and to grow again for 5-15 PDs, after which they
reached a second plateau. With 7 donors out of 8, we observed
a second emergence. These second emerging cells appeared
more transformed than the initial post-senescent ones. We
first believed these cells were immortalized and named them
ImKs for immortal keratinocytes. These cells, however,
underwent up to 60 PDs but then stopped and died. Neither
ImKs nor PS emerging cells showed any resumption of
telomerase activity, which was even lower than in young cells.
We estimated the emergence frequency of the first wave to
10-5-10-2, and that of the second wave to 10-5-10-4. These
frequencies are considerably higher than the frequency of
immortalization of SV40-transformed human fibroblasts that
is 10-7 (Shay & Wright, Exp Cell Res 184: 109; 1989).
Initiation and Evolution
of Epithelial Cancer
Corinne ABBADIE
PR2 - Univ Lille 1
Group Members :
Pourtier Albin, MCU Univ Lille 1
Bouali Fatima, Technician IPL
Vercamer Chantal, Engineer CNRS
Deruy Emeric, PhD Student IPL/Région
Malaquin Nicolas, PhD Student CNRS/Région
Post-senescent emerging keratinocytes are transformed
and tumorigenic
PS emerging keratinocytes look partly transformed, some
clones having kept an epithelioid morphology, and some
having acquired a fibroblastoid morphology associated with
a tendency to scatter. The expression of both involucrin and
keratin 14, two markers of keratinocyte differentiation, was
found to increase with senescence and to decrease again with
emergence. The expression of E-cadherin, a marker of
epithelial cell-cell adhesion, is also slightly reduced in PS
emerging cells. We performed DNA microarrays to compare
the transcriptomes of PS emerging keratinocytes with that of
young ones. Amongst the 50 most up-regulated and the 50
most down-regulated genes in PS emerging cells, 15 turn out
to be linked to adhesion or migration, 6 to cytoskeleton
structure or dynamics, 9 to senescence, oxidative stress, or
DNA damage, 9 to cell cycle progression or cell death, and 10
to diverse cancer-related pathways. Hence, about 50% of the
genes whose expression changes in emerging keratinocytes
are relevant to a transformed state.
We also investigated the karyotypic status and tumorigenic
potential of emerging cells. PS emerging cells displayed no
karyotypic aberrations but ImKs displayed many, mainly
translocations. Yet when injected into nude mice, both ImKs
and PS emerging cells generated disseminated skin lesions,
which revealed to be, after anatomopathological analysis,
hyperplasias and carcinomas.
Taken together, these results suggest that the cell populations
present at both growth plateaus yield partially transformed and
moderately tumorigenic emerging cells, with an apparent
progression on the pathway to transformation from emergence
wave to the next. Post-senescence emerging keratinocytes
might thus provide a good model for investigating the earliest
steps in the generation of tumour cells. These results were
published in Cancer Research (2009, 69, 7917-7925).
As a result of time and cumulative divisions in vitro as in vivo,
normal cells enter an irreversible non-proliferative state
termed replicative or cellular senescence that is thought to
contribute to organism aging. Both telomere shortening and
cumulative oxidative damage were shown to contribute to
senescence, probably acting at different degrees according to
proliferation index, cell type or environment. Because of its
associated cohort of damages and irreversible cell-cycle arrest,
senescence is commonly considered as a tumour-suppressor
mechanism that stops the proliferation of genetically altered
cells, i.e. potentially cancerous. However, the incidence of the
most frequent cancers in humans, carcinomas, exponentially
increases with age; the phenotypes of progeroid syndromes
are often associated with an increase in tumour incidence, and
inversely, when aging is delayed by caloric restriction, the
cancer incidence decreases (Martien & Abbadie, An N Y Acad
Sci 1119: 51; 2007). How to explain this positive link between
aging and tumorigenesis if, at the cell level, senescence is a
tumour-suppressor mechanism?
Our work during the four last years aimed to establish the
molecular links between senescence and neoplastic
transformation. We took advantage of the use of cell culture
models of human skin keratinocytes and fibroblasts whose
behaviour in vitro regarding senescence and neoplastic
transformation differs.
In contrast to skin fibroblasts, skin keratinocytes are able
to spontaneously escape senescence
When put into in vitro culture, normal human skin fibroblasts
undergo an exponential growth phase followed by an
irreversible growth plateau that can last several months.
Therefore, the concept of senescence as a tumor suppressor
phenomenon perfectly applies to this cell type. We have
observed that normal human skin keratinocytes behave in
vitro very differently. As fibroblasts, they first undergo an
exponential growth phase followed by a growth plateau
where they display the characteristics of senescent cells,
including increased cell size, polynucleation, accumulation of
vacuoles and various damaged components, senescenceassociated beta-Galactosidase (SA-b-Gal) activity, decreased
PCNA expression and increased p16 and p21 expression. After
a few days to 3 weeks at this plateau according to the donor,
several clones of small cells appeared spontaneously and
Mechanism of senescent keratinocyte death
The first step to understand the molecular mechanism of
emergence from senescence is to establish the mechanism of
senescence itself and the outcome of senescent cells in terms
of cell death.
We had previously established that senescence in
keratinocytes in mainly driven by the activation of a prooxidant patway involving NF-kappaB transcription factors and
one of their target genes, MnSOD. MnSOD (manganese
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Cancer
superoxide dismutase) encodes a redox enzyme which
transforms the toxic O2.- into H2O2, while catalase and
glutathione peroxidase (GPX) convert H2O2 into H2O. Neither
catalase nor GPX were co-upregulated with MnSOD during
senescence, leading to H2O2 accumulation (Bernard et al.,
Cancer Res 64: 472; 2004)
Based on the idea that keratinocytes should escape
senescence but also senescent cell death to emerge, we
investigated the mechanisms of senescent keratinocyte death.
We have shown that none apoptosis markers but several
hallmarks of macroautophagy, notably Beclin-1, appear with
the senescence of skin keratinocytes. The corpses that occur
at the senescent growth plateau display an accumulation of
vacuoles, lysotracker- and LC3-positive, hence of autophagic
nature. Three-methyladenine, an inhibitor of autophagosome
formation, but not zVAD, a caspase inhibitor, prevented
senescent-cell death. We concluded that senescent
keratinocytes do not die by apoptosis, but consequently to a
high macroautophagic activity that targets the main vital cell
components. These results were published in American
Journal of Pathology 2009, 174, 423-435). We have also several
data showing that the autophagic cell death pathway is
activated in senescent keratinocytes following their oxidative
damages (unpublished data).
2008
Furlan A, Vercamer C, Desbiens X, Pourtier, A.
Ets-1 triggers and orchestrates the malignant phenotype of mammary
cancer cells within their matrix environment.
J Cell Physiol, 2008, 215:782-793.
Molendi-Coste O, Mairesse J, Aubert N, Ghzili H, Abbadie C, Vaudry H,
Gonzales B, Anouar Y, Vieau D, Breton C, Laborie C.
Maternal perinatal undernutrition impairs chromaffin cells proliferation
in the postnatal rat.
Hormone Metab Res, 2008, 40:386-390.
2009
Gosselin K, Martien S, Pourtier A, Vercamer C, Ostoich P, Morat L,
Sabatier L, Duprez L, T'kint de Roodenbeke C, Gilson E, Malaquin N,
Wernert N, Slijepcevic P, Ashtari M, Chelli F, Deruy E, Vandenbunder B,
De Launoit Y, Abbadie C.
Senescence-associated oxidative DNA damage promotes the generation
of neoplastic cells.
Cancer Res, 2009, 69:7917-7925.
Am J Pathol, 2009, 174:423-435.
Humbert N, Martien S, Augert A, Da Costa M, Mauen S, Abbadie C,
de Launoit Y, Gil J, Bernard D.
A genetic screen identifies topoisomerase 1 as a regulator of senescence.
Cancer Res, 2009, 69:4101-4106.
In 2008 and 2009, our group also collaborated on different
research programs linked to apoptosis, senescence or
tumorigenesis :
- With C. Laborie, Unité de Neurosciences et Physiologie
Adaptatives, Lille 1 University, to characterize apoptotic cells
amongst adrenomedullary chromaffin cells in newborn rats
after maternal denutrition (Molendi-Coste et al., Horm
Metab Res 40: 386; 2008)
- With L. Sabatier, CEA Fontenay aux Roses, within an EC
granted project, to compare the effects of low dose
radiations on normal skin keratinocytes and fibroblasts
(work still in progress).
- With Albin Pourtier, who has join our team in 2006, and
achieve part of his previous project in our team regarding
the role of Ets1 in tumorigenesis (Furlan et al, J Cell Physiol,
2008, 215, 782-93)
- With N. Prevaskaya, Univ Lille 1, regarding the role in
cationic channel TRPV2 in prostate cancer cell migration
(Monet et, Biochim Biophys Acta, 2009, 1793, 528-39; Monet
et al, Cancer Research, 2010, 70, 1225-1235)
- With David Bernard in the lab, regarding karyotypic and
chromosomic damages associated with senescence
(Humbert, Cancer Research, 2009, 69, 4101-4106; Humbert
et al, EMBO J, 2010, 29, 376-386)
Monet M, Gkika D, Lehen'kyi V, Pourtier A, Vanden Abeele F,
Bidaux G, Juvin V, Rassendren F, Humez S, Prevarsakaya N.
Lysophospholipids stimulate prostate cancer cell migration via TRPV2
channel activation.
Sahin A, Vercamer C, Kaminski A, Fuchs T, Florin A, Hahne JC, Mattot V,
Pourtier-Manzanedo A, Pietsch T, Fafeur V, Wernert N. 2009
Ets 1 target genes.
Int J Oncol, 2009, 34:377-389.
2010
Deruy E, Gosselin K, Vercamer C, Martien S, Bouali F, Slomianny C,
Bertout J, Bernard D, Pourtier A, Abbadie C.
MnSOD upregulation induces autophagic programmed cell death in
senescent keratinocytes.
PlosOne, 2010, 5:e12712.
HumbertN, Navaratnam N, Augert A, Da Costa M, Martien S, Wang
J, Martinez D, Abbadie C, Carling D, de Launoit Y, Gil J, Bernard D.
Regulation of ploidy and senescence by the AMPK-related kinase NUAK1.
EMBO J, 2010, 29:376-386.
Publications
PhD
2007
Alessandro FURLAN
Directeur : A. Pourtier
Université de Lille
Decembre 2007
Martien S, Abbadie C.
Acquisition of oxidative damage during senescence : the first step towards
carcinogenesis.
Annals New York Acad Sci, 2007, 1119:51-63.
Sebastien MARTIEN
Directeur : C. Abbadie
Decembre 2009
Zdanov S, Bernard D, Debacq-Chainiaux F, Martien S, Gosselin K,
Vercamer C, Chelli F, Toussaint O, Abbadie C.
Normal or stress-induced fibroblast senescence involves COX-2 activity.
Exp Cell Res, 2007, 313:3046-3056.
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Cancer
Effect of H- PV on human cancer cells ex vivo
We developed a method allowing testing the oncolytic effect
of the H- PV on organotypic tumors. The oncolytic effect is
calculated thanks to the calibration previously performed with
a GFP-HeLa cell line. On these cells, 3-5 days of H1- PV are
sufficient to kill 95% (moi = 1) to 99% (moi = 5) of the GFPlabeled cells. More than 120 freshly excised tumors are
enzymatically and mechanically dissociated and cultured for
3-7 days in Petri dishes in the presence (moi = 1 or 5) or not of
H-1 PV. Previous reports show on a small number of samples
that these kinds of freshly excised tumors are killed by H-1 PV
(Van Pachterbeke et al., Int J Cancer 55: 672; 1993 - Van
Pachterbeke et al., Eur J Cancer 33: 1648; 1997). In very close
collaboration with the anticancer hospital « Centre Oscar
Lambret » of Lille, we showed that it is possible to obtain a
predictive test of the effect of H-1 PV on 15-20 day cultured
tumors. On a series of human breast tumors, 37% showed a
massive destruction of the cancer cells. On a first series of 8
sarcomas, 5 responded positively. These encouraging results
enable us to pursue this study on larger series of tumors.
Parvovirus :
Oncosuppression
and Gene Therapy
Dominique STEHELIN
DRCE - CNRS
Viruses are able to kill human cancer cells selectively (Alemany
et al., Nature Biotech 18: 723; 2000 – the review in Oncogene
24[52]; 2000). Although most of these viruses have negative
effects on the infected hosts, the autonomous H-1 parvovirus
(H-1 PV) is an oncolytic virus without any associated
pathology in human healthy adults and thus appears to be a
promising anticancer therapy alternative (Geletneky et al., J
Veterinary Med 52 : 327; 2005). It specifically and exclusively
replicates in a large number of transformed cell lines,
ultimately killing them. H-1 PV has thus three main properties:
• Oncotropism; i.e. specific of cancer cells.
• Oncolysis. After replication, H-1 PV destroys the cancer cells.
• Chain reaction. Destroyed cells liberate H-1 PV virions ready
to infect other cancer cells.
H-1 PV is also active in vivo on human cancer cells injected in
SCID mice. In this model, sufficient amount of H-1 PV is able
to cure the induced tumors.
Development of a GMP H-1 PV to further inject to patients
For this purpose, we had to develop a stable and highly pure
virus, without any animal contaminants. To stabilize the H-1
PV strain used in our experiments, we developed a molecular
sub-clone initially inserted in a bacterial plasmid. This DNA
was amplified and transfected in cultured cells in order to
obtain huge amounts of viral stocks of high quality. However,
to circumvent the problem of the presence of animal proteins
within the prepared virus, we selected and stabilized a nonadherent human cell line that can be maintained and
transfected without serum.
We have also developed an upstream process for H-1 PV
production based on a new technique, well adapted to cell
culture in suspension in a highly controlled disposable
Bioreactor (Wawe). We plan to develop a chromatographybased down-stream process and have contacted a
Biotech-industry (Henogen) to scale-up both upstream and
downstream processes. Henogen has already produced a
Development Cell Bank that will be tested by Texcell for
biological safety (viral contamination…) and a Development
Virus Seed (25-liter scale Production). These GLP made viruses
will be used for toxicity studies on immunocompetent rats
and for in vivo tests on the efficiency of H-1 PV on different
types of tumors induced in murine models.
Fundamental aspects of H-1 PV lysis mechanisms
H-1 PV life cycle highly depends on its non-structural protein
1 (NS1) whose expression is controlled by the parvoviral P4
promoter. NS1 indeed plays a key role in mechanisms such as
viral DNA replication and capsid proteins production and is
known to be the main effector of H-1 PV cytotoxicity on
transformed cells (Caillet-Fauquet et al., Embo J 9: 2989; 1990).
However, NS1 expression itself is not sufficient to induce hostcell death in normal cells. Three main steps have actually to
be achieved before H-1 PV-induced toxicity occurs in infected
transformed cells: (i) P4 promoter activation, (ii) NS1 activation
by specific post-translational modifications and (iii) interplay
between activated NS1 and cellular targets. These last two
years, we initiated a study on the molecular characterization
of the P4 promoter. For this purpose, we used a reporter
plasmid containing the P4 promoter mutated or not in the
different “consensus” core sites. Although H-1 PV P4 promoter
contains two putative NF-Y binding sites (Y1- and Y2-boxes)
located within the left-end palindrome of the viral genome,
we showed by transactivation assays that mutation of Y2-box
induces a dramatic decrease of P4 activity whereas the
suppression of Y1-box has no such significant effect. This is
correlated to the activity of the virus since Y2-box disruption
is linked to an extensive delay of release of progeny virions
from infected cells, possibly because of decreased NS1
expression. Interestingly, simultaneous mutations of both Y1and Y2-boxes affect not only the release of progeny virions
but also their production, which suggests that both Y boxes
act in synergy for NS1 expression and play a crucial role in H1 PV life cycle (Richard et al., XIIth Parvovirus Workshop
Abstract Book; 2008).
Publications
2010
Muharram G, Le Rhun E, Loison I, Wizla P, Richard A, Martin N,
Roussel A, Begue A, Devos P, Baranzelli MC, Bonneterre J,
Caillet-Fauquet P, Stehelin D.
Parvovirus H-1 induces cytopathic effects in breast carcinoma-derived
cultures.
Breast Cancer Res Treat, 2010, 121:23-33.
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Cleavage of Met by the presenelin-dependent regulated
intramembrane proteolysis (PS-RIP)
We also showed that Met is constitutively processed by the
Presenelin-dependent regulated intramembrane proteolysis
(PS-RIP). This proteolytic process involves sequential cleavages
by a membrane metalloprotease within the extracellular
region of Met and by the gamma-secretase complex (the
presenelin being the catalytic core) within its transmembrane
domain. The intracellular fragment generated by these
cleavages is extremely labile, through proteasome
degradation. Thus, these cleavages induce a constitutive
degradation of Met which reduces its half-life. Consequently,
this proteolytic process prevents accumulation of Met at the
membrane and its activation in absence of ligand (Foveau et
al., Mol Biol Cell, 20:2495-2507;2009).
Signalling, Apoptosis
and Cancer
David TULASNE
CR1 - Inserm
Group Members :
Chotteau-Lelièvre Anne, MCU Univ Lille 1
Lemière Arnaud, Engineer
Muharram Ghaffar, Postdoctoral fellow CNRS
Kherrouche Zoulika, Engineer IPL
Leroy Catherine, Assistant Engineer CNRS IPL
Damour Isabelle, Technician IPL
Ancot Frédéric, PhD Student MENRT
Ladam Franck, PhD Student MENRT
Lefebvre Jonathan, PhD Student MENRT
Therefore, our data revealed novel regulation of the Met
receptor by proteolytic cleavages, which act independently of
ligand-stimulation and extent biologic responses induced by
the receptor. In addition, deregulation of these proteolytic
processes could favour tumorigenis through deregulation of
the survival/apoptosis balance or overexpression of Met. In
this scientific context, we created in January 2008 an
emerging team, called «Signalling, Apoptisis and Cancer».
Regulation of the Met receptor
functions through proteolytic
cleavages
Pleiotropic functions of the ligand activated full length Met
The Hepatocyte growth factor/Scatter factor (HGF/SF), the
specific ligand of the tyrosine kinase receptor Met, induces
multiple biological responses including the proliferation,
invasion and morphogenesis of epithelial cells, acts as an
angiogenic factor in endothelial cells and has chemoattractant
and neurotrophic activities in different types of neurons.
HGF/SF is also a survival factor, which protects a number of
cell types against cell toxicity and apoptosis. Targeted
disruption of either hgf or met genes enlightens an essential
role of the HGF/SF-Met system during the development of the
placenta, liver, muscles and neurons (Birchmeier et al., Nat Rev
Mol Cell Biol, 4: 915; 2003). Deregulated HGF/SF-Met signalling
is involved in tumour development and progression in
particular by promoting invasion and metastasis in numerous
types of cancers (Migliore et al., Eur J Cancer, 44: 641; 2008).
Many groups study the signalling pathways and the biological
responses induced by the full length Met activated by its
ligand. However, various fragments or truncated Met
receptors have been observed, but their functional
involvements were not further characterised.
Publications
Met is reaped by Caspases
Although the HGF/SF-Met system is involved in cell survival,
we have shown that Met is cleaved by caspases, which are
crucial proteases for apoptosis. Indeed, in absence of ligand
and upon stress, Met is sequentially cleaved within its Cterminal extremity and within its juxtamembrane region.
Importantly, these cleavages lead to the generation of a
cytoplasmic fragment of 40 kDa (p40 Met), which is itself able
to promote apoptosis. Therefore, we have demonstrated that
in stress condition the Met survival receptor is converted by
caspase cleavages to an apoptotic factor able to regulate the
survival/apoptosis balance (Tulasne et al., Mol Cell Biol, 24:
10328; 2004 - Foveau et al., Cell Death Differ, 14: 752; 2007 Deheuninck et al., BBRC, 367: 573; 2008 - Tulasne et al., Cell
Death Differ, 15: 427; 2008).
Deheuninck J, Foveau B, Goormachtigh G, Leroy C, Ji Z, Tulasne D,
Fafeur V.
Caspase cleavage of the MET receptor generates an HGF interfering
fragment.
2007
Foveau B, Leroy C, Ancot F, Deheuninck J, Ji Z, Fafeur V, Tulasne D.
Cell Death Differ, 2007, 14:752-764.
Ji Z, Degerny C, Vintonenko N, Deheuninck J, Foveau B, Leroy C,
Coll J, Tulasne D, Baert JL, Fafeur C.
ubiquitinylation.
Oncogene, 2007, 26:395-406.
Trolet J, Fafeur V, Ben Younes A, Dissous C.
Int J Parasitol, 2007,14:1539-1550.
2008
Firlej V, Ladam F, Brysbaert G, Dumont P, de Launoit Y, Benecke A,
Chotteau-Lelièvre A.
Reduced tumorigenesis in mouse mammary cancer cells following
inhibition of either of the distinct Pea3 and Erm transcription programs.
J Cell Sci, 2008, 121:3393-3402.
Tulasne D, Foveau B.
The shadow of death on the MET tyrosine kinase receptor.
Cell Death Differ, 2008,15:427-434.(review)
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Cancer
2009
Pharmacochemistry
and Proteomics
Foveau B, Ancot F, Leroy C, Petrelli A, Reiss K, Vingtdeux V,
Giordano S, Fafeur V, Tulasne D.
Down-regulation of the Met receptor tyrosine kinase by presenilindependent regulated intramembrane proteolysis.
Mol Biol Cell, 2009, 20:2495-2507.
Patricia MELNYK
PU - Univ Lille 2
Villeret V, Tulasne D, Fafeur V.
1001 inhibits its caspase-dependent cleavage.
Cell Signal, 2009, 21:1455-1463.
The aim of our group is to design and synthesize candidatedrug in the field of neurodegenerative (Alzheimer’s disease),
neurologic and parasitic (Malaria) diseases. We are combining
structurally based rational design and parallel synthesis of
focused libraries. During the last years, in accordance with the
general axis of the unit, we have started a program based on
the design of c-Met inhibitors and their vectorisation. In
parallel, H. Drobecq conducts a plateform of mass
spectrometry – biomolecules analysis. He is thus engaged in
a lot of research projects with national and international
collaborations and is co-author of the publications.
Ancot F, Foveau B, Lefebvre J, Leroy C, Tulasne D.
Proteolytic cleavages give receptor tyrosine kinases the gift of ubiquity.
Oncogene, 2009, 28:2185-2195.
2010
Cazet A, Lefebvre J, Adriaenssens E, Julien S, Bobowski M,
Grigoriadis A, Tutt A, Tulasne D, Le Bourhis X, Delannoy P.
GD3 synthase expression enhances proliferation and tumor growth of
MDA-MB-231 breast cancer cells through c-met activation.
Mol Cancer Res, 2010, Oct 1.
New antimalarial compounds (collaboration Pr P. Grellier,
EA3335, MNHN, Paris)
Chloroquine (CQ) was a mainstream drug in the fight against
Plasmodium falciparum, but its efficacy is eroded by the
emergence of resistant parasites. We have developed
compounds able to accumulate into the food vacuole of the
parasite, inhibit the heme polymerisation more efficiently
than CQ. These compounds showed better antimalarial
properties in vitro and in some cases in vivo (Ryckebusch et al.,
Bioorg Med Chem Lett 15: 297; 2005). In all series, 1,4-bis(3aminopropyl)piperazine linker provided better activity upon
CQ-resistant strain (Deprez-Poulain et al., Comb Chem High
Throughput Screen 8: 39; 2005). Several series of amodiaquine
analogs, devoided of the 4’-phenol substituent, which seem
responsible of the toxicity, were synthesized (Delarue-Cochin
et al., Eur J Med Chem 43: 252; 2008 – Delarue-Cochin et al.,
Eur J Med Chem DOI: 10.1016/j.ejmech.2007.11.003) and
brought a new light on the SAR of the 4-anilinoquinolines
family. We also designed and analysed amodiaquine and
amopyroquine analogs, bearing new functionalities in 4’position : amine (Paunescu et al., Med Chem 2008, in press),
alkyle and aryle groups thanks to a Suzuki reaction (Paunescu
et al., Tetrahedron 63: 12791; 2007) and obtained two potent
compounds. In parallel, we also explored other type of
compounds, leading to the synthesis of the glyoxylhydrazone
type library of 80 compounds (Ryckebusch et al., Bioorg Med
Chem Lett 14: 4439; 2004) and focused library of iron chelating
acylhydrazones from salicylic aldehyde (Melnyk et al., Bioorg
Med Chem Lett 16: 31; 2006).
Choul-Li S, Leroy C, Leprivier G, Laitem C, Tulasne D, Aumercier M.
Caspase cleavage of Ets-1 p51 generates fragments with transcriptional
dominant-negative function.
Biochem J, 2010, 426:229-241.
PhD
Bénedicte FOVEAU
Directeur : D. Tulasne
Novembre 2007
HDR
Anne CHOTTEAU
HDR
Septembre 2009
Sigma-1 ligands as neuroprotector agents (collaborations
Dr T. Maurice, Inserm U710 Montpellier ; Pr C. Vaccher,
EA4034 Lille ; Pr R. Bordet, EA1046 Lille)
Sigma-1 protein and its effectors, endogenous agonists or
antagonists, are implicated in physiological desadaptions
leading to addiction. Previously we showed the affinity of
1,2,3,4-tetrahydroisoquinoline-hydantoine (Tic-hydantoïne)
mixed structure for σ 1 receptors. We have continued this
research program working on the synthesis methodologies
(Charton et al., Tetrahedron Lett 45: 7081; 2004), TicHyd
structure (Charton et al., Bioorg Med Chem Lett 15: 4833; 2005)
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Cancer
or amino side-chain (Cazenave-Gassiot et al., Bioorg Med
Chem Lett 15: 4828; 2005) modification. Nanomolar, selective
and non toxic agonists have shown a high potential for
cocaine weaning and are under development.
and controlled method of dextran lipidation allowing the
preparation of grafted nanoparticules was developed (Richard
et al., Bioconjugate Chem 2008, in press). In parallel, the design
of intracellular inhibitors combining both pharmacomodulation of published c-Met inhibitors and
chemoinformatic was undertaken to allow the search of new
pharmacophores. The main therapeutic application concerns
drug delivery to tumor sites.
Anti Alzheimer compounds (collaborations Dr A.
Delacourte, Inserm U837 Lille ; Pr C. Vaccher and Pr P.
Odou, EA4034 Lille)
Alzheimer’s disease (AD) is a frequent neurodegenerative
disorder in the aged population, provoking the complete loss
of cognitive functions and dementia. There is no cure and
symptomatic treatments available have marginal effects on
patients and on the evolution of the disease. APP protein is in
the heart of AD. One of its metabolites, the neurotoxic Aβ
peptide, aggregates to form amyloid plaques according to the
amyloidogenic pathway. The other competing way, the
neurotrophic pathway, prevents the formation of Aβ peptide
and provides APP fragments with neurotrophic and important
neurophysiological functions : sAPPalpha and AICD. Their
absence could stimulate Tau pathology, degenerating process
frequently encountered during aging and leading to AD.
Results obtained by A. Delacourte et al. from a human brain
library showed that sAPPalpha and AICD decreased during AD.
In a same way, familial forms of AD rely more on AICD decrease
than Aβ modifications. All recent publications underlined the
importance of APP loss of function in AD.
We have started a program aiming on slowing the
degradation of important APP fragments by the endosomelysosome system. We take advantage of our expertise of
antimalarial compounds and chloroquine analogs. This last
compound accumulates into the acidic food vacuole of the
parasite and inhibits heme polymerisation. A library of several
hundred of compounds was available in the lab and was
screened on a cellular test. Hits were optimized and a new
family of compounds (named MSBD) was patented (WO 2006
051489). Most potent derivatives increase APP-CTF alpha and
AICD fragment 10 times, leading to a parallel decrease in Aβ
secretion of 80%.
Our lead compound crosses the BBB, shows no toxicity on
mouse model, no cardio- or geno-toxicity and provides an
interesting ADME-Tox profile. This project is funding by an
“Aide au Transfert” Oseo grant.
In a parallel way, we have explored structurally different
compounds with potent pharmacophores and good theorical
physicochemical properties. We have thus identified and
developed a new family which is under patenting. This project
is funding by an ANR grant.
AlzProtect, a start-up company, was created to develop these
compounds through preclinical and clinical phases. Our
company was laureate of the “Concours national d’aide à la
création d’entreprises innovantes” in 2007.
Chemical strategy :
Firstly, our strategy was to propose original structures which
possess a central pharmacophore (pyrrole-indolin-2-one
structure present in many active molecules) and side chains
differently functionalized in order to improve the contacts
with ATP pocket of the MET TKR. The following process of lead
optimization was supported by computational chemistry
efforts. Ligand candidates are first aligned atop the
pharmacophore model extracted from the K252a binding
models (with receptor atoms accounted for by excluded
volumes) and then docked in to the MET active site based on
this alignment. Final interaction maps are realised with
ligdock. Currently, we have synthesised a chemical library
(around 40 compounds).
Secondly, we approached a rational design by an in silico
Virtual Screening (VS) of several drug libraries to identify
original and selective inhibitors of c-MET. VS is part of the early
stages of the drug discovery process. The main purpose is to
search efficiently small molecules in large databases and to
find a restricted number of them that have the highest
probability to be active. Docking and VS was realised in
collaboration with D. Horvath (UMR CNRS 7177).
Targeting strategy :
In parallel, we used lipid nanocapsules belong to the
generation of stealth nanovectors as TK inhibitor drug carriers.
Packaged into a nanocontainer, the drug is protected from
chemical and enzymatic degradation. These colloidal carriers,
in the nanometer size range, are produced using a solventfree process with biocompatible excipients. They are made up
of an oily core surrounded by a hydrophilic surfactant (PEG660
and functionalised polysaccharides). We have developed here
an efficient and convenient method to anchor a
polysaccharide onto the surface of lipid nanocapsules. This
method could be useful for decorating lipid particles with
functionalized polysaccharide with the aim of c-MET receptor
targeting.
This work, which corresponds to 2 years activity on a new
topic, was supported by the “Ligue Régionale contre le Cancer”
(15 k€).
C-Met inhibitors
In collaboration with team 6 « Cancer Biology and Chemistry
», our aim is to design and synthesize intracytoplasmic
inhibitors of c-Met tyrosine kinase activity and vectorise them
specifically to cancer cells thanks to lipid nanoparticules
decorated with functionalised dextrans. Depending on their
functionalised level, these polymers could selectively interact
with protein bearing heparin binding domain, such as c-Met
and HGF/SF. This was demonstrated in the case of PDGF-BB
growth factor, and confirmed in the lab using polysaccharide
microarrays (Carion et al., Chem Biochem 7: 817; 2006). A new
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Cancer
2007
Paunescu E, Susplugas S, Boll E, Varga R, Mouray E, Grosu I,
Grellier P, Melnyk P.
Replacement of 4’-hydroxy group of Amodiaquine and Amopyroquine by
aromatic and aliphatic substituants : Synthesis and Antimalarial Activity
ChemMedChem, 2009, 4:549-561.
Biet F, Angela de Melo Marques M, Grayon M, Xavier da Silveira EK,
Brennan PJ, Drobecq H, Raze D, Vidal Pessolani MC, Locht C,
Menozzi F.D.
Mycobacterium smegmatis produces an HBHA homologue which is
not involved in epithelial adherence.
Toussaint M , Delair B , Foulon C, Lempereur N, Vaccher C,
Maurice T, Melnyk P.
ic hydantoin sigma-1 agonist : pharmacological characterization on
cocaine-induced stimulant and appetitive effects.
Eur Neuropsychopharmacol, 2009, 19:504-515.
Publications
Dezitter X, Hammoudi F, Belverge N, Deloulme JC, Drobecq H,
Masselot B, Formstecher P, Mendy D, Idziorek T. 2007
Proteomics unveil corticoid-induced S100A11 shuttling in keratinocyte
differentiation.
PhD
Alexandre BARRAS
Directeur : N. Dupont
PhD « Chemistry »
Septembre 2009
Host H, Drobecq H, Locht C, Menozzi F.
FEMS Microbiol Lett, 2007, 268:144-150.
Marion TOUSSAINT
Directeur : P. Melnyk
PhD « Chemistry »
Septembre 2009
Medina-Palazon C; Gruffat H, Mure F, Filhol O, Vingtdeux-Didier V,
Drobecq H, Cochet C, Sergeant N, Sergeant A, Manet E.
Protein Kinase CK2 phosphorylation of EB2 regulates its function in the
production of EBV infectious viral particles
J. Virol, 2007, 81:11850-11860.
Paunescu E, Matuszak N, Melnyk P.
Suzuki coupling reaction as the key step for the synthesis of 4’-substituted
analogues of Amodiaquine.
Tetrahedron, 2007, 63:12791-12810.
Vingtdeux V, Hamdane M, Loyens A, Gelé P, Drobecq H, Begard S,
Galas MC, Delacourte A, Beauvillain JC, Buée L, Sergeant N.
Alkalizing drugs induce accumulation of Amyloid Precursor Protein
byproducts in luminal vesicles of multivesicular bodies.
J. Biol. Chem, 2007, 282:18197-18205.
2008
Delarue-Cochin S, Paunescu E, Maes L, Mouray E, Sergheraert C,
Grellier P, Melnyk P.
Synthesis and Antimalarial Activity of New Analogues of Amodiaquine
Eur J Medi Chem, 2008, 43:252-260.
Paunescu E, Susplugas S, Boll E, Vargas RA, Mouray E, Grellier P,
Melnyk P.
Synthesis and Antimalarial Activity of New Amino Analogues of
Amodiaquine and Amopyroquine
Med Chem, 2008, 4:407-425.
Richard A, Barras A, Ben Younes A, Dupont N, Melnyk P.
Minimal chemical modification of reductive end of dextran to produce
an amphiphilic polysaccharide able to incorporate onto lipid
nanocapsules
Bioconjug Chem, 2008, 19:1491-1495.
Venna VR, Deplancke D, Melnyk P, Bordet R.
Neuroprotective and antidepressant-like effects of LC 03/55, a novel
sigma-1 receptor ligand
Fund Clin Pharmacol, 2008, 22:1.
2009
Barras A, Mezzetti A, Richard A, Lazzaroni S, Roux S, Melnyk P,
Betbeder D. Dupont N.
Formulation and characterization of polyphenol-loaded lipid
nanocapsules Int
J Pharmaceut, 2009, 379:270-277.
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Laboratory
of Toxicology
previously EA2690, from january 2010
member of the « Equipe d’Accueil »
«Impact de l’environnement chimique sur
la santé humaine - Institut Pasteur de Lille
Daniel MARZIN
Contact :
00 33 3 20 87 79 75
[email protected]
Group members :
Le Curieux Frank, PU Univ Lille 2
Marzin Daniel, PU Univ Lille 2
Nesslany Fabrice, Assistant IPL
Merhi Maysaloun, Postdoctoral fellow
Platel Anne, PhD Student
Brient Alizée, Master 2 Student Univ Lille 2
Keys words :
Mutagenesis. Genotoxicity. Carcinogenesis. Risk assessment.
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7-Hydroxy-5-methyl-3-phenyl-6,7,8,9-tetrahydropyrido[3’,2’:
4,5]imidazo[1,2-a]pyrimidin-5-ium chloride (PhIP-M1) is a
newly identified intestinal microbial metabolite from the food
carcinogen 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP). Although the mutagenic potential of the
endogenous N-hydroxy PhIP derivate has been reported, the
risks associated with PhIP-M1 have not yet been explored. In
this work, the cytotoxic and genotoxic effects originating from
PhIP-M1 were assessed in the epithelial intestinal Caco-2 cell
line. A dose-dependent increase in DNA damage (quantified
by the alkaline comet assay) was observed after 3 h in the 50–
200 µM range. Moreover, PhIP-M1 induced apoptosis and cell
cycle arrest. Because these PhIP-M1-induced genomic and
cellular events may contribute to the carcinogenicity of PhIP,
the potency of the colon microbiota to bioactivate PhIP must
be included in future risk assessments.
A - Research Report
Genotoxicity of natural products
Aloe-emodin (AE) and derivatives may be present as
undesired components co-extracted during extraction of
plants containing anthraquinonic derivatives. AE is a wellknown in vitro mutagen, but up to now it failed to induce any
clear in vivo genotoxic activity in the chromosome aberration
assay in rat bone marrow or the in vivo/in vitro UDS test in liver.
However, the two target organs noted during rodent
carcinogenicity studies with danthron and emodin, two other
well-known anthraquinone derivatives, are the colon and the
kidney. Therefore, the choice of the organs for testing the
genotoxicity of AE, i.e. bone marrow and liver, may be
considered inadequate to demonstrate a possible in vivo
genotoxic activity. In this context, the in vivo mouse comet
assay was performed on both isolated kidney and colon
cells in order to demonstrate a possible organospecific
genotoxicity after oral administration. AE induced primary
DNA damage in the liver and in the kidney underlining an in
vivo genotoxic mechanism of action. Furthermore, AE induced
a clear genotoxic activity both in the Salmonella typhimurium
and in the in vitro micronucleus assay in TK6 human
lymphoblastoid cells in the absence as well as in the presence
of metabolic activation. As no significant variation in the
genotoxic activity of AE was noted when using either liver or
kidney S9-mix, it seems that no quantitatively and/or
qualitatively specific renal metabolism occurs. The kidney may
be a target organ of AE as it is the major route of excretion. AE
present in plant extracts should be considered as an in vivo
genotoxin and this property should be taken into account in
the risk assessment for human exposure.
Genotoxic potential of soils
A set of soil samples was collected from areas considered as not
contaminated by a known industrial activity : urban samples
(collected in cities), suburban samples (collected in villages),
agricultural samples, and forest or natural samples. The organic
extract of each soil sample was submitted to the micromethod
Ames test on Salmonella typhimurium strain TA98, with and
without a metabolic activation S9 system. A mutagenic activity
was observed in the extract of all 11 urban soil samples, and of
10 of the 15 suburban samples. On the other hand, the extract
of only one of the 7 agricultural samples studied induced
mutations, and none of the 18 natural or forest-soil samples
investigated produced mutagenic extracts. These findings
might indicate the crucial influence of the diffuse pollution
originating from different human activities on the mutagenic
potency of urban soil samples, and they make it possible to
classify the soils according to their mutagenic potency. No clear
correlation was found between the mutagenicity detected in
soil extracts and the measured polycyclic aromatic hydrocarbon
(PAH) content of the soils investigated.
Methylchavicol (or estragole), a natural flavouring substance
present in tarragon, was demonstrated as a genotoxic
chemical in the in vitro UDS test in cultured rat hepatocytes
and in the in vivo UDS test in hepatocytes of exposed rats.
Deep-frozen tarragon was clearly less genotoxic than
methylchavicol at equivalent dose levels, and desiccated
tarragon was negative. Both forms of tarragon tested in vitro
have the ability to decrease significantly the genotoxicity of
methylchavicol added to the culture medium. The decrease
may be attributed to antimutagenic properties of tarragon
leaves and/or to adsorption of methylchavicol, which would
decrease its bioavailability. Desiccated tarragon powder was
not genotoxic in the in vivo UDS test when administered up
to the maximum dose of 6.25 g/kg bw (18.75 mg/kg bw of
methylchavicol). In vivo, desiccated tarragon did not show
antimutagenic properties, because it did not decrease the
genotoxicity of methylchavicol administered at high
concentrations. Considering the low exposure level at the
maximum daily tarragon consumption, the rapid
detoxification and excretion in humans and the no-genotoxiceffect-level of methylchavicol by the oral route when given to
rats as tarragon leaves, a high margin of exposure exists. We
can conclude that tarragon consumption presents no
genotoxic risk to humans.
The DNA damage and cytotoxicity induced by CdCl2 solutions
and soils anthropogenically contaminated with heavy metals
was studied using the micronucleus (MN) test. Vicia faba, a
plant model widely used in liquid exposure assays, was
adapted for a direct exposure to a solid phase. In addition, the
MN assay was adapted to a new wild plant system, the white
clover (Trifolium repens). The results obtained after exposure
to CdCl2 solutions confirmed that Vicia faba root cells were a
sensitive model, and revealed that Trifolium repens root cells
were not appropriate for the detection of micronuclei
(although they were highly sensitive to the cytotoxic effect of
CdCl2 ). Concerning the results observed after direct exposure
to contaminated soils (solid phase exposure), the MN
frequency scores in Vicia faba root cells were increased in a
statistically significant and dose-related manner compared to
the control plants. Regarding Trifolium repens root cells, this
solid phase exposure confirmed that this model is not
appropriate for use in the micronucleus assay.
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Threshold of DNA oxidative damage
development 2010
The concept of threshold in genotoxic toxicology, its use in
risk assessment and its acceptance by scientists still under
debate. In this context, our study showed that DNA-oxidizing
agents exhibited non-linear dose-responses in the TK6 human
lymphoblastoid cells. No-Observed-Effect-Levels (NOELs)
could be defined with the micronucleus test and the
thymidine kinase gene mutation assay, i.e. for biomarkers of
effect, but not with the comet assay, i.e. for a biomarker of
exposure. Moreover, gene expression profiles were
investigated and revealed that the nature and the efficiency
of the activated cellular and molecular mechanisms depend
on the dose and the duration of exposure. TK6 cells triggered
several interacting signalling cascades and cellular functions
in response to oxidative stress and DNA damage (induction of
antioxidant defences, inflammatory and immune responses,
lipid metabolism, DNA repair, cell cycle control, cell death...).
Genotoxicology of nanoparticles :
Genotoxicity of nanoparticles : characterisation of crucial
physical and chemical properties and development of an
in vitro model of the superior aerial tract applicable to the
study of the hazard of particles of the urbano-industrial
environnement (project within the Institute of Research
on Industrial Environnement, IRENI)
The aim of the project is to characterise (1) the mechanisms
involved in the endocytosis of nanoparticles on the human
respiratory epithelium and (2) assess the influence of key
parameters (such as particle size or surface charge) on the
genotoxicity induced. We use a model of spheric porous
nanoparticles the parameters of which can be well-mastered,
and that can be marked and followed in the different cell
compartments. Different cell lines originating from the human
respiratory tract will be exposed to the model particles.
Moreover, the influence of the presence in the culture medium
of proteins will be investigated. The genotoxic effects are
assessed using two in vitro assays : the comet assay (detecting
primary DNA damage) and the micronucleus assay
(demonstrating chromosomal aberrations). In vivo geno toxicity assays may be performed in order to confirm the
validity of the data obtained in vitro.
Use of the comet assay to demonstrate the organospecificity of genotoxicants
The ability of the in vivo Comet Assay (pH>13) to distinguish
(1) genotoxicity vs cytotoxicity; (2) genotoxic vs nongenotoxic (epigenetic) carcinogens, was investigated in
Sprague-Dawley male rats by studying five known renal
genotoxic carcinogens acting through diverse mechanisms of
action (streptozotocin, aristolochic acids, 2-nitroanisole,
potassium bromate and cisplatin), two rodent renal epigenetic
carcinogens (D-limonene and cyclosporine) and two
nephrotoxic compounds (streptomycin and indomethacin).
Animals were treated once with the test compound by the
appropriate route of administration and genotoxic effects
were measured at the two sampling times of 3-6 hours and
22-26 hours after treatment. Under these experimental
conditions, the in vivo rodent Comet Assay demonstrated a
good sensitivity and a good specificity (all the 5 renal
genotoxic carcinogens were clearly detected in at least one
sampling time). In return, epigenetic and nephrotoxic
compounds failed to induce any significant increase in DNA
migration. In conclusion, the in vivo rodent Comet Assay
performed on isolated kidney cells could be used as a tool to
investigate the genotoxic potential of a test compound if
neoplasic/preneoplasic changes occur after subchronic or
chronic treatments in order to determine the role of
genotoxicity in tumor induction. Moreover, the epigenetic
carcinogens and cytotoxic compounds displayed clear
negative responses in this study. These results allow excluding
a DNA direct-acting mechanism of action and can thus
suggest that a threshold exists. Therefore, the current in vivo
rodent Comet assay may help to elucidate an epigenetic
mechanism and thus, to undertake a risk assessment
associated to the human use depending on the exposure
level.
Safety evaluation of manufactured nanomaterials by
characterization of their potential genotoxic hazard
(European Joint Action NANOGENOTOX)
The aim of the Joint Action is highly relevant in the current
context of nanotechnology and in particular, regarding the
human exposure to manufactured nanomaterials (MNs). It
targets crucial items identified in the programme providing a
genuine European dimension as 17 institutions from 11
Member States (MS), including new MS, are involved. The
methods and means implemented include 4 scientific Work
Packages with the aim of :
(i) strengthening, expanding and sharing the knowledge
required for hazard, exposure and overall risk assessment of
MNs, in particular by building a strategy able to generate
relevant and reliable data for EU public health authorities, (ii)
accelerating the exploitation of existing data and the exchange
of best practices in risk assessment and management, thus
minimising the potentially harmful long-term effects of MNs,
(iii) promoting the establishment of a robust methodology to
screen potentially genotoxic MNs. Using these results, a ring test
for the relevant assays is performed to check for possible
genotoxicity as alternative techniques to animal experimentation. In vivo assays are conducted to characterise the
toxicokinetics of selected MNs and validate the in vitro data.
MEGacity Aerosols chemical characterization and TOXicity
(MEGATOX) - ANR 2009-2011
The aim of the project is to characterize the chemical
composition and toxicity of particulate pollution in three
contrasted cities (in Europe, France, Paris; Asia, China, Xi’an;
Africa, Burkina Faso, Ouagadougou) with a special attention to
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the ultra-fine fraction (nanoparticles). Chemical mass balance
and source apportionment studies will be completed by more
specific source profile characterization and will be linked to
short-term and long-term biological effects on human
respiratory epithelial and endothelial cells. This multidisciplinary
task involves 8 partner-laboratories situated in France (7) and
in the USA (1). Five of the laboratories work on the collection
and the physical and chemical characterization of the three
different collected fractions (10-1; 1-0.1; <0.1 µm) while three
partners study the biological effects induced after exposure to
the particles. Our laboratory uses the comet assay in order to
assess the genotoxic effects induced on 16HBE, a human
bronchial epithemium cell line.
industrial waste dumps (leachate studied as such or after
treatment). The genotoxicity assays implemented will be one
in vitro assay (micromethod Ames test), and two in vivo assays
on the rat after oral administration of contaminated soil (bone
marrow micronucleus assay, and comet assay on 2 organs).
Publications
2007
Kirkland D, Pfuhler S, Tweats D, Aardema M, Corvi R, Darroudi F,
ElhajoujiA, Glatt H, Hastwell P, Hayashi M, Kasper P, Kirchner S,
Lynch A, Marzin D, Maurici D, Meunier JR, Müller L, Nohynek G,
Parry J, Parry E, Thybaud V, Tice R, van Benthem J, Vanparys P,
White P.
How to reduce false positive results when undertaking in vitro
genotoxicity testing and thus avoid unnecessary follow-up animal tests :
Report of an ECVAM Workshop.
Mutat Res, 2007, 628:31-55.
Heavy metals, Intestinal Ecotoxicology & in vivo (bio)Remediation : Natural & designer Probiotics versus
Xenobiotics in basal and pathological contexts (MELODIEREVE) - ANR 2010-2012
The aim of the project is to address toxicological aspects of
heavy metal (HM) exposure on the intestinal ecosystem, with
main consideration on the central role of the gut flora in
maintaining intestinal homeostasis. The final goal is to
propose a strategy for the manipulation of the microbiota,
using selected lactic acid bacteria and yeasts, as a preventive
and therapeutic approach to counteract the negative impacts
of HM contaminants. A preliminary screening of a wide
collection of natural microbial food-grade strains able to cope
with HM toxicity (native HM binding capacity, anti-oxidative
potential, anti-genotoxic and anti-inflammatory properties)
will be performed. The genotoxicity of 2 HM, i.e. lead (Pb) and
cadmium (Cd), on human enterocytes Caco2 cells will be
assessed using the Comet assay, a genotoxicity test able to
identify DNA damage such as single or double DNA strand
breaks (SSB or DSB), alkali-labile sites, DNA-DNA / DNA-protein
cross-linking and SSB associated with incomplete excision
repair sites. Caco2 cells are exposed to a range of Cd or Pb
concentrations, with and without each of the 5 strains (chosen
for their capacity to resist to HM and ROS), in order to assess
the possible preventive effects of these strains. This first step
will allow defining the best candidates for use as HM
detoxification tool. Then, for the best candidate, the mice in
vivo comet assay will be carried out on several organs in both
the Pb and the Cd models.
Marzin D.
Notion of threshold in mutagenesis : implications for mutagenic and
carcinogenic risk assessment.
Ann Pharm Fr, 2007, 65:404-414.
Nesslany F, Zennouche N, Simar-Meintières S, Talahari I, NKiliMboui EN, Marzin D.
In vivo Comet assay on isolated kidney cells to discriminate genotoxic
carcinogens from epigenetic carcinogens or cytotoxic compounds.
Mutat Res, 2007, 630:28-41.
Thybaud V,Aardema M, Clement J, Dearfield K, Galloway S,
Hayashi M, Jacobson-Kram D, Kirkland D, MacGregor J, Marzin D,
Ohyama W, Schuler M, Suzuki H, E. Zeiger.
Strategy for genotoxicity testing : Hazard identification and risk
assessment in relation to in vitro testing.
Mutat Res, 2007, 627:41-58.
2008
Courty B, Le Curieux F, Belkessam L, Laboudigue A, Marzin D.
Mutagenic potency in Salmonella typhimurium of organic extracts of soil
samples originating from urban, suburban, agricultural, forest and
natural areas.
Mutat Res, 2008, 653:1–5.
Nesslany. F., Simar-Meintières S., Watzinger M., Talahari I., Marzin D.
Nitrilotriacetic acid : an indirect in vitro and in vivo clastogen.
Environ Mol Mutagen, 2008, 49:439-452.
Markers of environmental genotoxicity (Comet assay,
micronucleus assay) for the assessment of the hazard
related to polycontaminated matrices (soil/air)
(MARGEEN) - AFSSET/ADEME 2010-2013
The objective of the project is to better assess the genotoxic
hazard induced by multiple and realistic exposures to complex
contaminated matrices (soil/air) and to develop diagnostic
tools able to detect the toxic fraction susceptible to induce
DNA damage.
To do so, the work will be performed according to two
successive phases. The aim of the first phase is to better
understand the mechanisms inducing DNA fragmentation
(are oxidative stress or synergistic effects involved ?). The
second phase is focused on in situ tests aiming at optimizing
the early detection of the genotoxic hazard already detected
in the first phase of the project.
Practically, our team will perform three genotoxicity assays on
four soil matrices.The studied soil matrices will contain either
a heavy metal (cadmium or lead) or a leachate obtained from
Vanhaecke L , Derycke L, Le Curieux F, Sofie Lust, Marzin D,
Verstraete W. Bracke M.
The microbial PhIP metabolite 7-hydroxy-5-methyl-3-phenyl-6,7,8,9
tetrahydropyrido[3,2:4,5]imidazo[1,2-a]pyrimidin-5-ium chloride (PhIP-M1)
induces DNA damage, apoptosis and cell cycle arrest towards Caco-2 cells.
Toxicology Letters, 2008,178:61-69.
2009
Gaudin J, Le Hegarat L, Nesslany F, Marzin D, Fessard V.
In vivo genotoxic potential of microcystin-LR, a cyanobacterial toxin,
investigated both by the unscheduled DNA synthesis (UDS) and the comet
assays after intravenous administration.
Environ Toxicol, 2009, 24:200-209.
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Khandoudi N, Porte P, Chtourou S, Nesslany F, Marzin D, Le Curieux F.
The presence of arginine may be a source of false positive results in the
Ames test.
Mutat Res, 2009, 679, 65-71.
Manier N., Deram A., Le Curieux F., Marzin D.
Comparison between new wild plant Trifolium repens and Vicia faba on
their sensitivity in detecting the genotoxic potential of heavy metal
solutions and heavy metal-contaminated soils.
Water, Air, & Soil Pollution, 2009, 202:343-352.
Nesslany F, Simar-Meintières S, Ficheux H, Marzin D.
Aloe-emodin-induced DNA fragmentation in the mouse in vivo comet
assay.
Mutat Res, 2009, 678:13-19.
Nesslany F, Parent-Massin D, Marzin D.
Risk assessment of consumption of methylchavicol and tarragon : the
genotoxic potential in vivo and in vitro.
Mutat Res, 2010, 696:1-9.
Platel A, Nesslany F, Gervais V, Marzin D.
Study of oxidative DNA damage in TK6 human lymphoblastoid cells by
use of the in vitro micronucleus test: Determination of No-Observed-Effect
Levels.
Mutat Res, 2009, 678:30-37.
PhD
Nicolas MANIER
Directeur de thèse : Frank Le Curieux
« Etude d'un nouveau modèle biologique végétal (Trifolium repens) en
écotoxicologie, applicable aux sols contaminés par les métaux lourds"
(ou éléments traces métalliques, aux choix).”
29 septembre 2008
Imed GARGOURI
Directeur de thèse : Daniel Marzin
« Evaluation de l'impact sanitaire des expositions professionnelles aux
solvants organiques dans l'industrie des colles et des chaussures de la
région de Sfax –Tunisie »
26 janvier 2009
Anne PLATEL
Directeurs de thèse : Marc Pallardy / Daniel Marzin
« Approches Génotoxiques et Transcriptomiques In Vitro pour la
Détermination de Seuils de Produits Induisant des Lésions Oxydatives à
l’ADN »
Université de paris XI
19 mars 2010
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Cardiovascular,
Metabolic and
Neurodegenerative
Diseases
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Public Health and
Molecular Epidemiology
of Aging-Related
Diseases
Inserm U 744
affilated to IFR 142
Philippe AMOUYEL
Contact :
00 33 3 20 87 77 10 / 77 62
[email protected]
Fiévet-Verrecas Nathalie, Engineer IPL
Marécaux Nadine, Engineer IPL
Codron Valérie, Technician IPL
Desmoucron Patricia, Technician IPL
Grupposo Marie-Catherine, Technician Inserm
Hermant Xavier, Technician IPL
Delbart Anne-Sophie, Administrative staff IPL
Peingnez Sabine, Administrative staff IPL
Steclebout Chantal, Administrative Staff IPL
Lombart Béatrice, Technician IPL
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3 Teams
• Public health and epidemiology of vascular diseases
Philippe AMOUYEL
Contact : 00 33 3 20 87 77 10 / 77 62 - [email protected]
• Search for the molecular determinants of cardiovascular diseases
via proteomic analysis and candidate gene approaches
Florence PINET
Contact : 00 33 3 20 87 72 15 - fl[email protected]
• Search for the molecular determinants of neurodegenerative diseases
via transcriptomic anc candidate gene approaches
Jean-Charles LAMBERT
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related to ageing in humans. Two major fields have been
studied : cardiovascular diseases and neurodegenerative diseases.
- This knowledge will enable us to suggest and develop
new strategies for the prevention and treatment of
these diseases.
A - Research Report
1 - Summary
Our objective consists in the gain of knowledge on the impact
of the environmental and genetic determinants in the
occurence, the evolution and the prevention and treatment
of two pathologies related to ageing : cardiovascular and
neurodegenerative diseases. The integration within the unit
of an epidemiological know-how and the control biology tools
(high throughput genomics, transcriptomics, proteomics,
bioinformatics) adjusted to the epidemiological approach,
allowed us to carry out this program.
One of the main specificities of this unit is the integration in
the same laboratory of various types of scientific and medical
know-how, from expertise in epidemiological methods and
clinical research to the use of analytical techniques
(genomics, transcriptomics, proteomics, bio-computing, cell
biology) to address certain functional aspects of
determinants identified.
Studies which have been conducted to date have enabled
us to answer a number of questions related to epidemiology
and public health of cardiovascular and neurodegenerative
diseases, and to suggest hypotheses on possible links
between these two affections.
The research we are developing is structured around three
closely dependent topics :
1. Public health and epidemiology of cardiovascular diseases.
The evolution of the coronary disease and its determinants
are studied. Focused around a morbidity registry,
descriptive and analytical studies are developed on
vascular risk factors in general population, in particular
metabolic, on secondary prevention and on 10 year
coronary risk.
Work performed by UMR Inserm 744 concentrate on 3 key
topics :
1. Public health and epidemiology of cardiovascular
diseases
2. Search for the molecular determinants of cardiovascular
diseases via proteomic analysis and candidate gene
approaches
2. Identification of the molecular determinants od
cardiovascular diseases. We identify by comparative
proteomics new determinants of vascular remodeling
(abdominal aorta aneurisms) and venticular remodeling
(heart failure) in clinical studies.
3. Search for the molecular determinants of neurodegenerative diseases via transcriptomic analysis and
candidate gene approaches
3. Identification of molecular determinants of neurodegenerative diseases. Initiated by our work on the
relationships between vascular and neurodegenerative
risk, we identify determinants of the impairment of
cognitive functions, in particular Alzheimer’s disease, by
genomic and transcriptomic approaches in case-control
and prospective studies.
The permanent interaction of these issues through the
analysis of concepts and technical and strategic complementarities is critical to the innovative character of the
scientific production of our unit.
In the twenty past years, we have produced or been involved
in nearly 50 clinical and epidemiologic research studies.
Considering the competences of researchers of the unit in
the fields of biology and genetics, most of these studies gave
rise to the creation of biological banks including samples
from more than 45,000 individuals, to tackle the study of the
biological and genetic determinants of these conditions. The
volume of these banks has led us to set up a biological
resource centre to maintain these samples following good
practice recommendations.
The analysis of the trends of coronary diseases allows us to
estimate their impact in public health. The identification of
proteinic markers of remodeling supports the development
of more targeted approaches of prevention. The candidate
genes identified in the decline of cognitive functions direct
towards original physiopathological pathways, orienting to
new treatment hypotheses. The knowledge generated by
this molecular epidemiology enables us to apprehend the
complexity of the determinism of multifactorial diseases.
II - Organisational chart of the Unit (as at 1
November 2008)
2 - Prior activity
I - Previous scientific project objectives
attained (in the past 4 years)
There have been changes in the unit staff - Doctor Mahmoud
Zureik left the unit in January 2007 to turn to respiratory
diseases epidemiology in Inserm unit 700 in Paris. He
therefore no longer is in charge of team 1, which now is
supervised by Philippe Amouyel. In such a context, the
project focused on the characterisation of sub-clinical
vascular alterations using new imaging techniques was
limited to studies that are underway.
The results described in the present paragraph reports the
achievement of the past 4 years. The main objective of this
unit is to :
- Gain knowledge on the impact of environmental and
genetic susceptibility factors in the development,
prevention and therapeutic treatment of diseases
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development 2010
Our project will be developed in the continuity of our
objectives of grain of knowledge on the impact of the
environmental and genetic determinants in the occurrence,
the evolution and the prevention and treatment of two major
ageing affections : cardiovascular and neurodegeneratives
diseases. Our project is organiszed according to three closely
interrelated axes :
1. Public health and epidemiology of cardiovascular diseases.
Our epidemiologic studies carried out on the cornary and
metabolic risk will be exploited in detail. The increase of life
expectancy and of cerebrovascular risk lead us to develop
a morbidity registry on stroke and a large European casecontrol study (4000 subjects) on cerebral artery dissection,
for which a whole genome analysis is scheduled.
2. Identification of molecular determinants of cardiovascular
diseases. The impact of the determinants of vascular and
ventricular remodeling we identified will be analyzed. We
will continue to implement our differential proteomic
techniques and will use these approaches in collaboration
on animal remodeling models. We will also apply our
approaches to the identification of markers of silent
cerebral ischaemia related to cardiac catheterization.
3. Identification of molecular determinants of neurodegenerative diseases. We will continue the genome analysis in
a large case-control study on Alzheimer’s disease (10 000
subjects) coupled to a European application study (10 000
other subjects). We will develop also functional analyses of
the markers we will identify.
These developments of adapted epidemiologic tools and
powerful molecular approaches will enable us to accelerate in
a competitive way the discovery of new determinants of the
diseases related to ageing and to open the way to new
preventive and therapeutic approaches.
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registers, which have been working in close cooperation for
more than twenty years, provide French data related to the trend
for cardiovascular diseases in the population. We also develop
cross-sectional epidemiological studies (population-based
investigation on a representative sample, clinical series) at
regular intervals and analytical studies (case-control &
prospective studies) to try to explain the trends observed in the
registry. These studies conducted by our teams have enabled us
to demonstrate or take part in the identification and assessment
of the impact of determinants for cardiovascular diseases.
Public Health
and Epidemiology
of Vascular Diseases
Philippe AMOUYEL
PU PH Lille 2
Group Members :
Dallongeville Jean, DR IPL
Dauchet Luc, AHU Lille 2
Dumont Julie, MCU Lille 2
Meirhaeghe-Hurez Aline, CR1 Inserm
Montaye Michèle, Médecin IPL
Goumidi Louisa, Research assistant Contrat UE
Beauchant Stéphanette, Médecin IPL
Cloez Anne, Infirmière IPL
Cottel Dominique, Médecin IPL
Cousin Béatrice, Infirmière IPL
Devoghelaere Christine, Médecin IPL
Dumont Marie-Pierre, Infirmière IPL
Graux Catherine, Médecin IPL
Lemaire Brigitte, Médecin IPL
Pygeire Marie, PhD Student Lille 2
Bodenant Marie, Master 2 Paris XI
Wyndels Karine, Master 2 Paris XI
Lecompte Sophie, Master 2 Student Lille 2
During our initial four-year project, we had set to ourselves as
objectives the completion of a number of epidemiological
studies, in particular :
1. a national, representative population-based investigation
on risk factors for the cardiovascular disease (Monalisa
investigation: National Arterial Risk Monitoring) following
the first two population-based investigations of the Monica
project (1985-87, 1995-97)
2. the validation of events 10 years later for the PRIME
prospective study on myocardial infarction
3. a European study on primary prevention of the coronary
disease (Euroaspire III investigation: EUROpean Action on
Secondary Prevention and Intervention to Reduce Events)
following the two previous ones (1995, 2000)
4. a longitudinal study on the acute coronary syndrome in
connection with the morbidity register
5. a case-control study on morbid obesity in adults
6. a case-control study on the risk of dissecting aneurysm of
the carotid
7. the development of a morbidity register for cerebrovascular diseases.
Key words :
Public heath. Molecular epidemiology. Aging. Cardiovascular
diseases (coronary heart disease, aneurysm, heart failure). Stroke.
Morbidity register. Risk factors. High throughout genomics.
Cardiovascular risk. Translational research. Prevention
Recruitments for the first three studies are over and findings
are now being worked upon. Recruitments are still underway
for the four last studies and data should be examined at the
end of 2009.
The objectives of this topic are :
- to gain more knowledge on the development of the
cardiovascular disease,
- to characterise its environmental and genetic determinants,
- to characterise sub-clinical vascular alterations using new
imaging techniques to estimate the cardiovascular risk.
Among the main preliminary findings obtained, the data of the
three French registers have shown that the average annual rates
of events for people aged 35 to 64 in France are 277/100,000 for
men and 54/100,000 for women (1997-2002) (Ruidavets et al,
2007; Montaye et al, 2007). Incidence rates are 240/100,000 for
men against 49/100,000 for women, and mortality rates reach
112/100,000 for men and 28/100,000 for women (Arveiler et al,
2007). In men, the standardised rates of coronary events have
tended to drop in Lille (-2.8% per annum) when they remained
stable in Strasbourg and have tended to increase in a nonsignificant way in Toulouse (+1.6% per annum). In women, the
analysis of trends does not show any significant change in rates,
whether centres may be considered separately or combined. For
both sexes, whatever the year, event rates are still higher in Lille
and Strasbourg than in Toulouse. The Euroaspire III study,
conducted in European coronary patients more than 6 months
after their events, has evidenced the increased prevalence of
obesity and overweight in the past 10 years. Finally, the initial
analyses of the Monalisa study have shown that in the global
population, the overweight level between 35 and 74 was higher
that regular estimates reported in France and that it affected two
thirds of men and 50% of women.
Descriptive, analytical and intervention epidemiology is required
to develop knowledge on the cardiovascular disease and
characterise its determinants. Part of our research efforts are
developed in the framework of the morbidity register of
ischemic cardiopathies in the Urban Community of Lille, which
has been keeping track of all myocardial infarction cases in a
population of approximately 1 million men and women aged 35
to 74, in a continuous and exhaustive manner since 1985. This
register, which is qualified by the “ Comité National de Registres
”, was initially created as part of the international monitoring
programme for cardiovascular diseases steered by the World
Health Organisation, MONICA project (Multinational
mONItoring of trends and determinants of CArdiovascular
diseases). To date, this programme still is the broadest
epidemiological study ever to have been launched on
cardiovascular diseases, the objective of which was to study the
trend of the incidence of cardiovascular diseases and
determinants which may account for such trend over at least 10
years. Thirty-eight registers in twenty-one countries were
involved in this study, including three located in France: “Urban
Community of Lille”, Bas-Rhin and Haute-Garonne. These three
During the past 4 years, we have also examined the impact of
certain environmental determinants on cardiovascular risk. Part
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of this work was focused on vascular risks related to the
metabolic syndrome and nutrition. The metabolic syndrome is
a clinical entity characterised by a combination of
cardiometabolic risk factors related to obesity (particularly in its
abdominal form) and insulin resistance. In the PRIME study, we
have evidenced an excess of coronary risk in subjects who met
existing definitions of the syndrome (Bataille et al, 2006). In the
MONICA population-based investigation, we could show a link
between the family history of early coronary disease and
metabolic syndrome suggesting that the latter could account
for part of the aggregation of coronary risk factors in families
(Dallongeville et al, 2006). This work is complemented by clinical
studies where we were able to assess the impact of the weight
loss induced by a hypo-caloric diet on the metabolic markers
of insulin resistance and on the appetite control hormones. We
demonstrated differentiated responses to test meals according
to lipid or carbohydrates content (Dallongeville et al, 2007;
Romon et al, 2006).
last study, we were able to demonstrate a link between the
increased carotid distensibility and risk of coronary disease in
the elderly (Leone et al, 2008), as well as the reduction in the
number of carotid plaques in women who regularly drink tea
(Debette et al, 2008). Several associations were also identified
between these sub-clinical vascular alterations and certain
genetic polymorphisms. For instance, a polymorphism of the
apolipoprotein E gene was associated with carotid plaques
(Debette et al, 2006) and several polymorphisms in the genes
of urea metabolism to the intima-media thickness variability
(Dumont et al, 2007a & 2007b).
The development and international scientific recognition of
these projects require the combination of a number of
conditions enabling the validation of results : sample size,
quality of recruitment, replication of results in various
populations, physiopathological hypotheses. This is why many
projects are conducted as part of international collaborative
efforts combining several teams to bring together several
hundreds or even thousands of subjects. The studies of the
register of coronary heart diseases are published in cooperation
with the other French centres at least, often with the Irish centre
of Belfast whose cardiovascular risk level is significantly higher
than the risk for France. The Euroaspire study results from
collaborative efforts with more than 20 European countries,
including 9 which were involved in the early stage of the study
in 1994, for trend analyses. Finally, our studies are also part of
larger consortiums to ensure sufficient power to answer certain
questions on vascular risk factors (The Fibrinogen Study
Collaboration, 2005) as well as in the Morgam project
(International pooling projectMOnica Risk, Genetics, Archiving
and Monograph) developed following the Monica programme.
Publications
We also explored the influence of genetic polymorphisms on
the risk of obesity, type-2 diabetes, or metabolic syndrome in
the MONICA population-based survey or in case-control studies
on obesity or type-2 diabetes. We could demonstrate that
APOA4 (Dallongeville et al, 2005), KLF2 (Meirhaeghe et al,
2006a) and perilipin genes (Meirhaeghe et al, 2006b) do not
seem to be involved in the regulation of body weight. In
contrast, we could show that a polymorphism of the CART gene
increases the risk of obesity (Guerardel et al, 2005) and
atherosclerosis risk (Vasseur et al, 2007). We have also found that
the polymorphisms of BNP (Meirhaeghe et al, 2007),
adiponectine (Vasseur et al, 2005), HNF-4 (Vaxillaire et al,
2005), KFL11 (Neve et al, 2005), KFL10 and SMAD7 genes
(Gutierrez-Aguilar et al, 2007) modulated the risk of type-2
diabetes and that a polymorphism of the LXR gene (liver X
receptor) is linked to higher circulating HDL-cholesterol rates
and a reduced risk of metabolic syndrome (Legry et al, 2008).
Finally, we have shown that a haplotype of the PPARG gene is
associated with a 2.3-fold increase in the metabolic syndrome
risk (Meirhaeghe et al, 2005) and that the Pro12Ala
polymorphism of the PPARG gene was a risk factor for
prematurity (risk multiplied by 2) in 2 independent cohorts of
Irish teenagers and young adults (Meirhaeghe et al, 2007).
2007
Appleton KM, Woodside JV, Yarnell JW, Arveiler D, Haas B,
Amouyel P, Montaye M, Ferrières J, Ruidavets JB, Ducimetière P,
Bingham A, Evans A ; for the PRIME Study Group.
Type A behaviour and consumption of an atherogenic diet : No
association in the PRIME study.
Appetite, 2007, 49:554-560.
Appleton KM, Woodside JV, Yarnell JW, Arveiler D, Haas B,
Amouyel P, Montaye M, Ferrières J, Ruidavets JB, Ducimetière P,
Bingham A, Evans A ; for the PRIME Study Group.
Depressed mood and dietary fish intake : Direct relationship or indirect
relationship as a result of diet and lifestyle ?
J Affect Disord, 2007, 104:217-223.
Boddaert J, Kinugawa K, Lambert JC, Boukhtouche F, Zoll J,
Merval R, Blanc-Brude O, Mann D, Berr C, Vilard J, Garabedian B,
Silvestre JS, Duyckaerts C, Amouyel P, Mariani J, Tedgui A, Mallat Z.
Evidence of a role for lactadherin in Alzheimer's disease.
Am J Pathol, 2007, 170:921-929.
Dallongeville J, Gruson E, Dallinga-Thie G, Pigeyre M, Gomila S, Romon M.
Effect of weight loss on the postprandial response to high-fat and highcarbohydrate meals in obese women.
Eur J Clin Nutr, 2007, 61:711-718.
The third aspect of this research program is the characterisation
of sub-clinical vascular alterations using non-invasive imaging
techniques to estimate the cardiovascular risk. These research
efforts were developed on the basis of measurements made via
carotid echography in two epidemiologic studies: the EVA study
(Assessment of Artery Ageing) and “Etude des Trois Cités”. In this
de Groote P, Mouquet F, Dallongeville J, Lamblin N, Bauters C.
The impact of the AMPD1 gene polymorphism on exercise capacity, other
prognostic parameters, and survival in patients with stable congestive
heart failure. A study on 686 consecutive patients.
Am Heart J, 2007, 153:e15.
112
Contents
De Henauw S, Gottrand F, De Bourdeaudhuij I, Gonzalez-Gross M,
Leclercq C, Kafatos A, Molnar D, Marcos A, Castillo M, Dallongeville J,
Gilbert CC, Bergman P, Widhalm K, Manios Y, Breidenassel C,
Kersting M, Moreno LA, on behalf of the HELENA Study Group.
Nutritional status and lifestyles of adolescents from a public health
prespective. The HELENA Project - Healthy Lifestyle in Europe by Nutrition
in Adolescence.
J Publ Health, 2007, 15:187-197.
Graham I, Atar D, Borch-Johnsen K, Boysen G, Burell G, Cifkova R,
Dallongeville J, et al.
European guidelines on cardiovascular disease prevention in clinical
practice: executive summary : Fourth Joint Task Force of the European
Society of Cardiology and Other Societies on Cardiovascular Disease
Prevention in Clinical Practice : Executive summary.
Eur Heart J, 2007, 28:2375-2414.
Dallongeville J, De Backer G, Ebrahim S, Gjelsvik B, HerrmannLingen C, Hoes A, Humphries S, Knapton M, Perk J, Priori SG,
Pyorala K, Reiner Z, Ruilope L, Sans-Menendez S, Op Reimer WS,
Weissberg P, Wood D, Yarnell J, Zamorano JL.
ESC Committee for Practice Guidelines. European guidelines on
cardiovascular disease prevention in clinical practice : Executive summary.
Atherosclerosis, 2007, 194:1-45.
Dumont J, Zureik M, Cottel D, Montaye M, Ducimetière P,
Amouyel P, Brousseau T.
Association of arginase 1 gene polymorphisms with the risk of myocardial
infarction and common carotid intima-media thickness.
J Med Genet, 2007, 44:526-531.
Dumont J, Zureik M, Bauters C, Grupposo MC, Cottel D,
Montaye M, Hamon M, Ducimetière P, Amouyel P, Brousseau T.
Association of OAZ1 Gene Polymorphisms With Subclinical and Clinical
Vascular Events.
Arterioscler Thromb Vasc Biol, 2007, 27:2120-2126
Guyant-Maréchal L, Rovelet-Lecrux A, Goumidi L, Cousin E,
Hannequin D, Raux G, Penet C, Ricard S, Macé S, Amouyel P,
Deleuze JF, Frebourg T, Brice A, Lambert JC, Campion D.
Variations in the APP promoter region and risk to Alzheimer’s disease.
Neurology, 2007, 68:632-633.
Emerging Risk Factors Collaboration, Danesh J, Erqou S, Walker M,
Thompson SG, Tipping R, Ford C, Pressel S, Walldius G, Jungner I,
Folsom AR, Chambless LE, Knuiman M, Whincup PH, Wannamethee SG,
Morris RW, Willeit J, Kiechl S, Santer P, Mayr A, Wald N, Ebrahim S,
Lawlor DA, Yarnell JW, Gallacher J, Casiglia E, Tikhonoff V, Nietert PJ,
Sutherland SE, Bachman DL, Keil JE, Cushman M, Psaty BM, Tracy RP,
Tybjaerg-Hansen A, Nordestgaard BG, Frikke-Schmidt R, Giampaoli S,
Palmieri L, Panico S, Vanuzzo D, Pilotto L, Simons L, McCallum J,
Friedlander Y, Fowkes FG, Lee AJ, Smith FB, Taylor J, Guralnik J,
Phillips C, Wallace R, Blazer D, Khaw KT, Jansson JH, Donfrancesco C,
Salomaa V, Harald K, Jousilahti P, Vartiainen E, Woodward M,
D'Agostino RB, Wolf PA, Vasan RS, Pencina MJ, Bladbjerg EM,
Jorgensen T, Moller L, Jespersen J, Dankner R, Chetrit A, Lubin F,
Rosengren A, Wilhelmsen L, Lappas G, Eriksson H, Bjorkelund C,
Cremer P, Nagel D, Tilvis R, Strandberg T, Rodriguez B, Bouter LM,
Heine RJ, Dekker JM, Nijpels G, Stehouwer CD, Rimm E, Pai J, Sato S,
Iso H, Kitamura A, Noda H, Goldbourt U, Salomaa V, Salonen JT,
Nyyssönen K, Tuomainen TP, Deeg D, Poppelaars JL, Meade T, Cooper
J, Hedblad B, Berglund G, Engstrom G, Döring A, Koenig W, Meisinger C,
Mraz W, Kuller L, Selmer R, Tverdal A, Nystad W, Gillum R, Mussolino M,
Hankinson S, Manson J, De Stavola B, Knottenbelt C, Cooper JA, Bauer
KA, Rosenberg RD, Sato S, Naito Y, Holme I, Nakagawa H, Miura H,
Ducimetière P, Jouven X, Crespo C, Garcia-Palmieri M, Amouyel P et al.
The Emerging Risk Factors Collaboration : analysis of individual data on
lipid, inflammatory and other markers in over 1.1 million participants in
104 prospective studies of cardiovascular diseases.
Eur J Epidemiol, 2007, 22:839-869.
Helbecque N, Cottel D, Hermant X, Amouyel P.
Impact of the matrix metalloproteinase MMP-3 on dementia.
Neurobiol Aging, 2007, 28:1215-1220.
Komajda M, Amouyel P, Johnson N, Bergougnoux L, Laperche T,
de Groote P, Jaillon P, Cohen-Solal A.
Comité scientifique d'étude de KEOPS et des investigateurs. Treating heart
failure with carvedilol in private practice (initiating treatment and followup at one year). The KEOPS study.
Arch Mal Coeur Vaiss, 2007, 100:818-826.
Lambert JC, Ferreira S, Gussekloo J, Christiansen L, Brysbaert B,
Slagboom PE, Cottel D, Petit T, Hauw JJ, Dekosky ST, Richard F,
Berr C, Lendon CL, Kamboh IM, Mann D, Christensen K,
Westendorp R, Amouyel P.
Evidence for the association of the S100beta gene with low cognitive
performance and dementia in the elderly.
Mol Psychiatry, 2007, 12:870-880.
Lambert JC, Amouyel P.
Genetic heterogeneity of Alzheimer's disease: Complexity and advances.
Psychoneuroendocrinology, 2007, 32 Suppl 1:S62-S70.
Meirhaeghe A, Sandhu MS, McCarthy MI, de Groote P, Cottel D,
Arveiler D, Ferrières J, Groves CJ, Hattersley AT, Hitman GA, Walker
M, Wareham NJ, Amouyel P.
Association between the T-381C polymorphism of the brain natriuretic
peptide gene and risk of type 2 diabetes in human populations.
Hum Mol Genet, 2007, 16:1343-1350.
Evans A, Marques-Vidal P, Ducimetière P, Montaye M, Arveiler D,
Bingham A, Ruidavets JB, Amouyel P, Haas B, Yarnell J, Ferrières J,
Fumeron F, Luc G, Kee F, Cambien F.
Patterns of alcohol consumption and cardiovascular risk in northern
ireland and france.
Ann Epidemiol, 2007, 17(5 Suppl):S75-80.
Meirhaeghe A, Boreham CA, Murray LJ, Richard F, Davey Smith G,
Young IS, Amouyel P.
A possible role for the PPARG Pro12Ala polymorphism in preterm birth.
Diabetes, 2007, 56:494-498.
practice: full text. Fourth Joint Task Force of the European Society of
Cardiology and other societies on cardiovascular disease prevention in
clinical practice : Full text.
Eur J Cardiovasc Prev Rehabil, 2007, 14 Suppl 2:S1-S113.
Montalescot G, Dallongeville J, Van Belle E, Rouanet S, Baulac C,
Degrandsart A, Vicaut E ; for the OPERA Investigators .
STEMI and NSTEMI : are they so different ? 1 year outcomes in acute myocardial infarction as defined by the ESC/ACC definition (the OPERA registry).
Eur Heart J, 2007, 28:1409-1417.
practice: executive summary. Fourth Joint Task Force of the European
Society of Cardiology and other societies on cardiovascular disease
prevention in clinical practice : Executive summary.
Eur J Cardiovasc Prev Rehabil, 2007, 14 Suppl 2:E1-E40.
Puig JG, Marre M, Kokot F, Fernandez M, Jermendy G, Opie L,
Moyseev V, Scheen A, Ionescu-Tirgoviste C, Saldanha MH,
Halabe A, Williams B, Mion D, Ruiz M, Hermansen K, Tuomilehto J,
Finizola B, Gallois Y, Amouyel P, Ollivier JP, Asmar R.
Efficacy of indapamide SR compared with enalapril in elderly
hypertensive patients with type 2 diabetes.
Am J Hypertens, 2007, 20:90-97.
113
Contents
Ruidavets JB, Bongard V, Dallongeville J, Arveiler D, Ducimetière P,
Perret B, Simon C, Amouyel P, Ferrières J.
High consumptions of grain, fish, dairy products and combinations of
these are associated with a low prevalence of metabolic syndrome.
J Epidemiol Community Health, 2007, 61:810-817.
Campagne F, Lambert JC, Dreses-Werringloer U, Vingtdeux V,
Lendon C, Campion D, Amouyel P, Lee AT, Gregersen PK, Davies P,
Marambaud P.
CALHM1 association with Alzheimer's disease risk response.
Cell, 2008, 135:994-996.
Troughton JA, Woodside JV, Young IS, Arveiler D, Amouyel P,
Ferrières J, Ducimetière P, Patterson CC, Kee F, Yarnell JW, Evans A.
Homocysteine and coronary heart disease risk in the PRIME study.
Chaix B, Ducimetière P, Lang T, Haas B, Montaye M, Ruidavets JB,
Arveiler D, Amouyel P, Ferrières J, Bingham A, Chauvin P.
Residential environment and blood pressure in the PRIME Study : is the
association mediated by body mass index and waist circumference ?
J Hypertens, 2008, 26:1078-1084.
Troughton JA, Woodside JV, Young IS, Arveiler D, Amouyel P,
Ferrières J, Ducimetière P, Patterson CC, Kee F, Yarnell JW, Evans A.
PRIME Study Group. Bilirubin and coronary heart disease risk in the
Prospective Epidemiological Study of Myocardial Infarction (PRIME).
Eur J Cardiovasc Prev Rehabil, 2007, 14:79-84.
Chapuis J, Hannequin D, Pasquier F, Bentham P, Brice A, Leber I,
Frebourg T, Deleuze JF, Cousin E, Thaker U, Amouyel P, Mann D,
Lendon C, Campion D, Lambert JC.
Association study of the GAB2 gene with the risk of developing
Alzheimer's disease.
Neurobiol Dis, 2008, 30:103-106.
Vasseur F, Guerardel A, Barat-Houari M, Cottel D, Amouyel P,
Froguel P, Helbecque N.
Impact of a CART promoter genetic variation on plasma lipid profile in a
general population.
Mol Genet Metab, 2007, 90:199-204.
Chapuis J, Moisan F, Mellick G, Elbaz A, Pasquier F, Hannequin D,
Lendon C, Campion D, Amouyel P, Lambert JC.
Association study of the NEDD9 gene with the risk of developing
Alzheimer's and Parkinson's disease.
Hum Mol Genet, 2008, 17:2863-2867.
Vicente-Rodriguez G, Libersa C, Mesana MI, Béghin L, Iliescu C,
Moreno Aznar LA, Dallongeville J, Gottrand F.
Healthy Lifestyle by Nutrition in Adolescence (HELENA). A New EU Funded
Project.
Therapie 2007; 62:259-270.
Chouraki V, Wagner A, Ferrières J, Kee F, Bingham A, Haas B,
Ruidavets J-B, Evans A, Ducimetière P, Amouyel P, Dallongeville J.
Waist circumference, smoking and 10 years coronary artery disease risk
in middle aged men : the PRIME study.
Amouyel P.
Epidémiologie des syndromes coronaires aigus.
In : Cardiologie et maladies cardiovasculaires.
Ed. Société Française de Cardiologie. Masson, 2007, 88-94.
Cieniewski-Bernard C, Mulder P, Henry JP, Drobecq H, Dubois E,
Pottiez G, Thuillez C, Amouyel P, Richard V, Pinet F.
Proteomic analysis of left ventricular remodelling in an experimental
model of heart failure.
J Proteome Res, 2008, 7:5004-5016.
Bongard V, Dallongeville J.
Diabète et Syndrome Métabolique.
In : Cardiologie et maladies cardiovasculaires. Ed. Société Française de
Cardiologie. Masson, 2007, 109-112.
Cieniewski-Bernard C, Acosta A, Dubois E, Lamblin N, Beseme O,
Chwastyniak M, Amouyel P, Bauters C, Pinet F.
Proteomic analysis in cardiovascular diseases.
Clin Exp Pharmacol Physiol, 2008, 35:362-366.
Dallongeville J.
Evaluation du risque cardiovasculaire.
In : Cardiologie et maladies cardiovasculaires. Ed. Société Française de
Cardiologie. Masson, 2007, 265-268.
Dallongeville J, Cottel D, Wagner A, Ducimetière P, Ruidavets JB,
Arveiler D, Bingham A, Ferrières J, Amouyel P, Meirhaeghe A.
The APOA5 Trp19 allele is associated with metabolic syndrome via its
association with plasma triglycerides.
BMC Med Genet, 2008, 9:84.
Dallongeville J, Dauchet L, Amouyel P.
Epidémiologie de l’Obésité Abdominale.
In : L’obésité abdominale, une maladie métabolique. Ed. Després JP.
Collection Pathologie, Science, Formation. Libbey J, EUROTEXT 2007, 5-28.
Dallongeville J, Bringer J, Bruckert E, Charbonnel B, Dievart F,
Komajda M, Pouchain D, Amouyel P.
Abdominal obesity is associated with ineffective control of cardiovascular
risk factors in primary care in France.
Diabetes Metab, 2008, 34:606-611.
2008
Biggio V, Renneville A, Nibourel O, Philippe N, Terriou L, Roumier C,
Amouyel P, Cottel D, Castaigne S, Dombret H, Thomas X, Fenaux P,
Preudhomme C.
Recurrent in-frame insertion in C/EBPalpha TAD2 region is a polymorphism without prognostic value in AML.
Leukemia, 2008, 22:655-657.
Dauchet L, Dallongeville J.
Fruit and vegetables and cardiovascular disease : epidemiological
evidence from the non-Western world.
Br J Nutr, 2008, 99:219-220.
Bruandet A, Richard F, Bombois S, Maurage CA, Masse I,
Amouyel P, Pasquier F.
Cognitive decline and survival in Alzheimer’s disease according to
education level.
Dement Geriatr Cogn Disord, 2008, 25:74-80.
Debette S, Courbon D, Leone N, Gariépy J, Tzourio C, Dartigues JF,
Barberger-Gateau P, Ritchie K, Alpérovitch A, Amouyel P,
Ducimetière P, Zureik M.
Tea consumption is inversely associated with carotid plaques in women.
Arterioscler Thromb Vasc Biol, 2008, 28:353-359.
Bruandet A, Richard F, Tzourio C, Berr C, Dartigues JF,
Alpérovitch A, Amouyel P, Helbecque N.
Haplotypes across ACE and the risk of Alzheimer's disease : The Three-City
Study.
J Alzheimers Dis, 2008, 13:333-339.
Debette S, Leone N, Courbon D, Gariépy J, Tzourio C, Dartigues JF,
Ritchie K, Ducimetière P, Amouyel P, Zureik M.
Calf circumference is inversely associated with carotid plaques.
Stroke, 2008, 39:2958-2965.
114
Contents
Dreses-Werringloer U, Lambert JC, Vingtdeux V, Zhao H, Vais H,
Siebert A, Jain A, Koppel J, Rovelet-Lecrux A, Hannequin D,
Pasquier F, Galimberti D, Scarpini E, Mann D, Lendon C, Campion D,
Amouyel P, Davies P, Foskett JK, Campagne F, Marambaud P.
A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels,
and Alzheimer's disease risk.
Cell, 2008, 133:1149-1161.
Troughton JA, Woodside JV, Yarnell JW, Arveiler D, Amouyel P,
Ferrières J, Ducimetière P, Patterson CC, Luc G ; on behalf of the
PRIME Study Group.
Paraoxonase activity and coronary heart disease risk in healthy middleaged males : The PRIME study.
Combris P, Amiot-Carlin M-J, Caillavet F, Causse M, Dallongeville J,
Padilla M, Renard C, Soler L-G.
Les fruits et légumes dans l'alimentation. Enjeux et déterminants de la
consommation. Série Expertise collective.
Éditeur Quae, Versailles, France. 2008.
Dupont A, Chwastyniak M, Beseme O, Guihot AL, Drobecq H,
Amouyel P, Pinet F.
Application of saturation dye 2D-DIGE proteomics to characterize
proteins modulated by oxidized low density lipoprotein treatment of
human macrophages.
J Proteome Res, 2008, 7:3572-3582.
2009
Acosta-Martin AE, Chwastyniak M, Beseme O, Drobecq H,
Amouyel P, Pinet F.
Impact of incomplete DNase I treatment on human macrophage
analysis.
Proteomics Clin Appl, 2009, 3:1-11.
Empana JP, Canoui-Poitrine F, Luc G, Juhan-Vague I, Morange PE,
Arveiler D, Ferrières J, Amouyel P, Bingham A, Montaye M, Ruidavets JB,
Haas B, Evans A, Ducimetière P, on behalf of The PRIME Study Group :
Contribution of novel biomarkers to incident stable angina and acute
coronary syndrome. The PRIME Study.
Eur Heart J, 2008, 29:1966-1974.
Helbecque N, Codron V, Cottel D, Amouyel P.
An apolipoprotein A-I gene promoter polymorphism associated with
cognitive decline, but not with Alzheimer's disease.
Ahluwalia N, Ferrières J, Dallongeville J, Simon C, Ducimetière P,
Amouyel P, Arveiler D, Ruidavets JB.
Association of macronutrient intake patterns with being overweight in a
population-based random sample of men in France.
Diabetes Metab, 2009, 35:129-136.
Legry V, Cottel D, Ferrières J, Deroide T, Amouyel P, Meirhaeghe A.
Association between liver X receptor alpha gene polymorphisms and
metabolic syndrome in French populations.
Int J Obes, (Lond) 2008, 32:421-428.
Allouch M, Zhong YZ, Riddell JW, Sabatier R, Hamon M.
Transradial coronary rotational atherectomy using 5-French guiding
catheters.
Chin Med J (Engl), 2009, 122:1356-1358.
Leone N, Ducimetière P, Gariépy J, Courbon D, Tzourio C, Dartigues JF,
Ritchie K, Alpérovitch A, Amouyel P, Safar ME, Zureik M.
Distension of the carotid artery and risk of coronary events.
The Three-City Study.
Amouyel P, Mismetti P, Langkilde LK, Jasso-Mosqueda G,
Nelander K, Lamarque H.
INR variability in atrial fibrillation : a risk model for cerebrovascular events.
Eur J Intern Med, 2009, 20:63-69.
Asplund K, Karvanen J, Giampaoli S, Jousilahti P, Niemelä M,
Broda G, Cesana G, Dallongeville J, Ducimetriere P, Evans A,
Ferrières J, Haas B, Jorgensen T, Tamosiunas A, Vanuzzo D,
Wiklund PG, Yarnell J, Kuulasmaa K, Kulathinal S; MORGAM Project.
Relative risks for stroke by age, sex, and population based on follow-up
of 18 European populations in the MORGAM Project.
Stroke, 2009, 40:2319-2326.
Lieb W, Zeller T, Mangino M, Götz A, Braund P, Wenzel JJ, Horn C,
Proust C, Linsel-Nitschke P, Amouyel P, Bruse P, Arveiler D, König IR,
Ferrières J, Ziegler A, Balmforth AJ, Evans A, Ducimetière P,
Cambien F, Hengstenberg C, Stark K, Hall AS, Schunkert H,
Blankenberg S, Samani NJ, Erdmann J, Tiret L.
Lack of association of genetic variants in the LRP8 gene with familial and
sporadic myocardial infarction.
J Mol Med, 2008, 86:1163-1170.
Barthélémy O, Beygui F, Vicaut E, Rouanet S, Van Belle E, Baulac C,
Degrandsart A, Dallongeville J, Montalescot G ; OPERA Investigators.
Relation of high concentrations of plasma carboxy-terminal telopeptide
of collagen type I with outcome in acute myocardial infarction.
Am J Cardiol, 2009, 104:904-909.
Moreno LA, Gonzalez-Gross M, Kersting M, Molnar D, de Henauw S,
Beghin L, Sjöström M, Hagströmer M, Manios Y, Gilbert CC,
Ortega FB, Dallongeville J, Arcella D, Wärnberg J, Hallberg M,
Fredriksson H, Maes L, Widhalm K, Kafatos AG and Marcos A,
on behalf of the HELENA Study Group.
Assessing, understanding and modifying nutritional status, eating habits
and physical activity in European adolescents : The HELENA (Healthy
Lifestyle in Europe by Nutrition in Adolescence) Study.
Public Health Nutr, 2008, 11:288-299.
Bensemain F, Hot D, Ferreira S, Dumont J, Bombois J, Maurage CA,
Huot L, Hermant X, Levillain E, Hubans C, Hansmannel F, Chapuis J,
Lemoine Y, Hauw JJ, Schraen S, Lemoine Y, Buée L, Berr C, Mann D,
Pasquier F, Amouyel P, Lambert JC.
Evidence for induction of the ornithine transcarbamylase expression in
Mol Psychiatry, 2009, 14:106-116.
Pinet F, Beseme O, Cieniewski-Bernard C, Drobecq H, Jourdain S,
Lamblin N, Amouyel P, Bauters C.
Predicting left ventricular remodeling after a first myocardial infarction
by plasma proteome analysis.
Proteomics, 2008, 8:1798-1808.
Beygui F, Montalescot G, Vicaut E, Rouanet S, Van Belle E, Baulac C,
Degrandsart A, Dallongeville J ; OPERA Investigators.
Aldosterone and long-term outcome after myocardial infarction :
A substudy of the french nationwide Observatoire sur la Prise en charge
hospitalière, l'Evolution à un an et les carRactéristiques de patients
présentant un infArctus du myocarde avec ou sans onde Q (OPERA) study.
Am Heart J, 2009, 157:680-687.
Sabouret P, Cacoub P, Dallongeville J, Krempf M, Mas JL, Pinel JF,
Priollet P, Steg G, Taminau D, Montalescot G ; for the REACH Registry
investigators.
REACH : International prospective observational registry in patients at
risk of atherothrombotic events. Results for the French arm at baseline
and one year.
Arch Cardiovasc Dis, 2008, 101:81-88.
Bokor S, Amouyel P, Dallongeville J.
Which measure of adiposity for primary care ?
Int J Clin Pract, 2009, 63:1270-1272.
115
Contents
Bruandet A, Richard F, Bombois S, Maurage CA, Deramecourt V,
Lebert F, Amouyel P, Pasquier F.
Alzheimer's disease with cerebrovascular disease and vascular dementia
: clinical features and course compared with Alzheimer's disease.
J Neurol Neurosurg Psychiatry, 2009, 80:133-139.
Debette S, Metso TM, Pezzini A, Engelter ST, Leys D, Lyrer P,
Metso AJ, Brandt T, Kloss M, Lichy C, Hausser I, Touzé E, Markus HS,
Abboud S, Caso V, Bersano A, Grau A, Altintas A, Amouyel P,
Tatlisumak T, Dallongeville J, Grond-Ginsbach C ; CADISP group.
CADISP-genetics : an International project searching for genetic risk
factors of cervical artery dissections.
Int J Stroke, 2009, 4:224-230.
Canoui-Poitrine F, Luc G, Juhan-Vague I, Morange PE, Arveiler D,
Ferrieres J, Amouyel P, Bingham A, Montaye M, Ruidavets JB,
Haas B, Evans A, Ducimetiere P, Empana JP; on behalf of The PRIME
Study Group.
Respective contribution of conventional risk factors and antihypertensive
treatment to stable angina pectoris and acute coronary syndrome as first
presentation of coronary heart disease. The PRIME Study.
Delhaye C, Sudre A, Lemesle G, Maréchaux S, Broucqsault D,
Hennache B, Bauters C, Lablanche JM.
Preprocedural high-sensitivity C-reactive protein predicts death or
myocardial infarction but not target vessel revascularization or stent
thrombosis after percutaneous coronary intervention.
Cardiovasc Revasc Med, 2009, 10:144-150.
Chapuis J, Boscher M, Bensemain F, Cottel D, Amouyel P, Lambert JC.
Association study of the paraoxonase 1 gene with the risk of developing
Neurobiol Aging, 2009, 30:152-156.
Deschaintre Y, Richard F, Leys D, Pasquier F.
Treatment of vascular risk factors is associated with slower decline in
Alzheimer’s disease.
Neurology, 2009, 73:674-680.
Chapuis J, Hot D, Hansmannel F, Kerdraon O, Ferreira S, Hubans C,
Maurage CA, Huot L, Bensemain F, Laumet G, Ayral AM, Fievet N,
Hauw JJ, DeKosky ST, Lemoine Y, Iwatsubo T, Wavrant-De Vrièze F,
Dartigues JF, Tzourio C, Buee L, Pasquier F, Berr C, Mann D,
Lendon C, Alpérovitch A, Kamboh MI, Amouyel P, Lambert JC.
Transcriptomic and genetic studies Identify IL-33 as a candidate gene for
Mol Psychiatry, 2009, 14:1004-1016.
Do HQ, Nazih H, Luc G, Arveiler D, Ferrières J, Evans A, Amouyel P,
Cambien F, Ducimetière P, Bard JM.
Influence of cholesteryl ester transfer protein, peroxisome proliferatoractivated receptor alpha, apolipoprotein E, and apolipoprotein A-I
polymorphisms on high-density lipoprotein cholesterol, apolipoprotein
A-I, lipoprotein A-I, and lipoprotein A-I:A-II concentrations : the
Prospective Epidemiological Study of Myocardial Infarction study.
Metabolism, 2009, 58:283-289.
Clément K, Stunff CL, Meirhaeghe A, Dechartres A, Ferrieres J,
Basdevant A, Boitard C, Amouyel P, Bougnères P.
In obese and non-obese adults, the cis-regulatory rs361072 promoter
variant of PIK3CB is associated with insulin resistance not with type 2
diabetes.
Mol Genet Metab. 2009; 96(3):129-132.
Dumont J, Meroufel D, Bauters C, Hansmannel F, Bensemain F,
Cottel D, Hamon M, Lambert JC, Ducimetière P, Amouyel P,
Zureik M, Brousseau T.
Association of ornithine transcarbamylase gene polymorphisms with
hypertension and coronary artery vasomotion.
Am J Hypertens, 2009, 22:993-1000.
Clergeau M-R, Hamon M, Morello R, Saloux E, Viader F, Hamon M.
Silent cerebral infarcts in patients with pulmonary embolism and a patent
foramen ovale : a prospective diffusion-weighted magnetic resonnance
imaging study.
Stroke, 2009, 40:3758-3762.
Emerging Risk Factors Collaboration, Erqou S, Kaptoge S, Perry PL,
Di Angelantonio E, Thompson A, White IR, Marcovina SM, Collins R,
Thompson SG, Danesh J.
Lipoprotein(a) concentration and the risk of coronary heart disease,
stroke, and nonvascular mortality.
JAMA, 2009, 302:412-423.
Coronary Artery Disease Consortium, Samani NJ, Deloukas P,
Erdmann J, Hengstenberg C, Kuulasmaa K, McGinnis R, Schunkert H,
Soranzo N, Thompson J, Tiret L, Ziegler A.
Large scale association analysis of novel genetic loci for coronary artery
disease.
Emerging Risk Factors Collaboration, Di Angelantonio E, Sarwar N,
Perry P, Kaptoge S, Ray KK, Thompson A, Wood AM, Lewington S,
Sattar N, Packard CJ, Collins R, Thompson SG, Danesh J.
Major lipids, apolipoproteins, and risk of vascular disease.
JAMA, 2009, 302:1993-2000.
Dallongeville J, Iribarren C, Ferrières J, Lyon L, Evans A, Go AS,
Arveiler D, F
ortmann SP, Ducimetière P, Hlatky MA, Amouyel P, Southwick A,
Quertermous T, Meirhaeghe A. Peroxisome proliferator-activated receptor
gamma polymorphisms and coronary heart disease.
PPAR Res, 2009, 2009:543746.
Empana JP, Dauvilliers Y, Dartigues JF, Ritchie K, Gariepy J,
Jouven X, Tzourio C, Amouyel P, Besset A, Ducimetiere P.
Excessive daytime sleepiness is an independent risk indicator for
cardiovascular mortality in community-dwelling elderly : the three city study.
Stroke, 2009, 40:1219-1224.
Ferrières J, Bongard V, Dallongeville J, Arveiler D, Cottel D, Haas B,
Wagner A, Amouyel P, Ruidavets JB.
Trends in plasma lipids, lipoproteins and dyslipidaemias in French adults,
1996-2007.
Dauchet L, Amouyel P, Dallongeville J ; Medscape.
Fruits, vegetables and coronary heart disease.
Nat Rev Cardiol, 2009, 6:599-608.
Debette S, Bevan S, Dartigues JF, Sitzer M, Lorenz M, Ducimetière P,
Amouyel P, Markus HS.
Fractalkine receptor/ligand genetic variants and carotid intima-media
thickness.
Stroke, 2009, 40:2212-2214.
Fibrinogen Studies Collaboration.
Measures to assess the prognostic ability of the stratified Cox proportional
hazards model.
Stat Med, 2009, 28:389-411.
116
Contents
Fibrinogen Studies Collaboration, Jackson D, White I, Kostis JB,
Wilson AC, Folsom AR, Wu K, Chambless L, Benderly M, Goldbourt
U, Willeit J, Kiechl S, Yarnell JW, Sweetnam PM, Elwood PC, Cushman
M, Psaty BM, Tracy RP, Tybjaerg-Hansen A, Haverkate F, de Maat MP,
Thompson SG, Fowkes FG, Lee AJ, Smith FB, Salomaa V, Harald K,
Rasi V, Vahtera E, Jousilahti P, D'Agostino R, Kannel WB, Wilson PW,
Tofler G, Levy D, Marchioli R, Valagussa F, Rosengren A, Wilhelmsen
L, Lappas G, Eriksson H, Cremer P, Nagel D, Curb JD, Rodriguez B,
Yano K, Salonen JT, Nyyssönen K, Tuomainen TP, Hedblad B,
Engström G, Berglund G, Loewel H, Koenig W, Hense HW,
Meade TW, Cooper JA, De Stavola B, Knottenbelt C, Miller GJ,
Cooper JA, Bauer KA, Rosenberg RD, Sato S, Kitamura A, Naito Y,
Iso H, Salomaa V, Harald K, Rasi V, Vahtera E, Jousilahti P, Palosuo T,
Ducimetiere P, Amouyel P, Arveiler D, Evans AE, Ferrieres J, JuhanVague I, Bingham A, Schulte H, Assmann G, Cantin B, Lamarche B,
Despres JP, Dagenais GR, Tunstall-Pedoe H, Lowe GD, Woodward M,
Ben-Shlomo Y, Davey Smith G, Palmieri V, Yeh JL, Meade TW,
Rudnicka A, Brennan P, Knottenbelt C, Cooper JA, Ridker P,
Rodeghiero F, Tosetto A, Shepherd J, Lowe GD, Ford I, Robertson M,
Brunner E, Shipley M, Feskens EJ, Di Angelantonio E, Kaptoge S,
Lewington S, Lowe GD, Sarwar N, Thompson SG, Walker M,
Watson S, White IR, Wood AM, Danesh J.
Systematically missing confounders in individual participant data metaanalysis of observational cohort studies.
Stat Med, 2009, 28:1218-1237.
Hachulla E, Carpentier P, Gressin V, Diot E, Allanore Y, Sibilia J,
Launay D, Mouthon L, Jego P, Cabane J, de Groote P, Chabrol A,
Lazareth I, Guillevin L, Clerson P, Humbert M; ItinérAIRSclérodermie Study Investigators.
Risk factors for death and the 3-year survival of patients with systemic
sclerosis : the French ItinérAIR-Sclérodermie study.
Rheumatology (Oxford), 2009, 48:304-308.
Fibrinogen Studies Collaboration.
Correcting for multivariate measurement error by regression calibration
in meta-analyses of epidemiological studies.
Stat Med, 2009, 28:1067-1092.
An age effect on the association of common variants of ACE with
Neurosci Lett, 2009, 461:181-184.
Gardette V, Bongard V, Dallongeville J, Arveiler D, Bingham A,
Ruidavets JB, Amouyel P, Haas B, Ducimetière P, Ferrières J.
Ten-year all-cause mortality in presumably healthy subjects on lipidlowering drugs (from the Prospective Epidemiological Study of
Myocardial Infarction [PRIME] prospective cohort).
Am J Cardiol, 2009, 103:381-386.
Helbecque N, Cottel D, Amouyel P.
Low-density lipoprotein receptor-related protein 8 gene polymorphisms
and dementia.
Hamon M, Rasmussen LH, Manoukian SV, Cequier A, Lincoff AM,
Rupprecht HJ, Gersh BJ, Mann T, Bertrand ME, Mehran R, Stone GW.
Choice of arterial access site and outcomes in patients with acute coronary
syndromes managed with an early invasive strategy : The Acuity trial.
Eurointervention, 2009, 5:115-120.
Hamon M, Coutance G.
Transradial intervention for minimizing bleeding complications in
percutaneous coronary intervention.
Am J Cardiol, 2009, 104(5 Suppl):55C-59C. Review.
Hansmannel F, Lendon C, Pasquier F, Dumont J, Hannequin D,
Chapuis J, Laumet G, Ayral AM, Galimberti D, Scarpini E, Campion D,
Amouyel P, Lambert JC.
Is the ornithine transcarbamylase gene a genetic determinant of
Alzheimer's disease ?
Neurosci Lett, 2009, 449:76-80.
Jolly SS, Amlani S, Hamon M, Yusuf S, Mehta SR.
Radial versus Femoral Access for Coronary Angiography or Intervention
and the Impact on Major Bleeding : A Systematic Review and Metaanalysis of Randomized Trials.
Am Heart J, 2009, 157:132-40.
Goumidi L, Spengler D, Cottel D, Wagner A, Ducimetière P,
Ruidavets JB, Legry V, Arveiler D, Bingham A, Ferrières J,
Amouyel A, Meirhaeghe A.
Study of the genetic variability of ZAC1 (PLAGL1) in French populationbased samples.
J Hypertens, 2009, 27:314-321.
Karvanen J, Silander K, Kee F, Tiret L, Salomaa V, Kuulasmaa K,
Wiklund PG, Virtamo J, Saarela O, Perret C, Perola M, Peltonen L,
Cambien F, Erdmann J, Samani NJ, Schunkert H, Evans A, MORGAM
Project.
The impact of newly identified loci on coronary heart disease, stroke and
total mortality in the MORGAM prospective cohorts.
Genet Epidemiol, 2009, 33:237-246.
Hachulla AL, Launay D, Gaxotte V, de Groote P, Lamblin N, Devos P,
Hatron PY, Beregi JP, Hachulla E.
Cardiac magnetic resonance imaging in systemic sclerosis : a crosssectional observational study of 52 patients.
Ann Rheum Dis, 2009, 68:1878-1884.
Konoshita T, Kato N, Fuchs S, Mizuno S, Aoyama C, Motomura M,
Makino Y, Wakahara S, Inoki I, Miyamori I, Pinet F.
Genetic variant of the renin-angiotensin system and diabetes influences
blood pressure response to angiotensin receptor blocker.
Diabetes care, 2009, 32:1485-1490.
Hachulla E, de Groote P, Gressin V, Sibilia J, Diot E, Carpentier P,
Mouthon L, Hatron PY, Jego P, Allanore Y, Tiev KP, Agard C,
Cosnes A, Cirstea D, Constans J, Farge D, Viallard JF, Harle JR,
Patat F, Imbert B, Kahan A, Cabane J, Clerson P, Guillevin L,
Humbert M ; Itinér AIR-Sclérodermie Study Group.
The three-year incidence of pulmonary arterial hypertension associated
with systemic sclerosis in a multicenter nationwide longitudinal study in
France.
Arthritis Rheum, 2009, 60:1831-1839.
Kotseva K, Wood D, De Backer G, De Bacquer D, Pyörälä K, Keil U ;
EUROASPIRE Study Group.
Cardiovascular prevention guidelines in daily practice : a comparison of
EUROASPIRE I, II, and III surveys in eight European countries.
Lancet, 2009, 373:929-940.
Hachulla E, Bervar JF, Launay D, Lamblin N, Perez T, Mouthon L,
de Groote P, Tillie-Leblond I, Humbert M.
[Dyspnea upon exertion in systemic scleroderma : from symptom to
etiological diagnosis] [Article in French]
Presse Med, 2009, 38:911-926.
Kotseva K, Wood D, Backer GD, Bacquer DD, Pyörälä K, Keil U ;
EUROASPIRE Study Group.
EUROASPIRE III : a survey on the lifestyle, risk factors and use of
cardioprotective drug therapies in coronary patients from 22 European
countries.
Eur J Cardiovasc Prev Rehabil, 2009, 16: 121-137.
117
Contents
Labayen I, Ruiz JR, Vicente-Rodríguez G, Turck D, Rodríguez G,
Meirhaeghe A, Molnár D, Sjöström M, Castillo MJ, Gottrand F,
Moreno LA.
Early life programming of abdominal adiposity in adolescents ; The
HELENA study.
Diabetes Care, 2009, 32:2120-2122.
Mouquet F, Lamblin N, de Groote P.
Indication for bromocriptine in peripartum cardiomyopathy.
Eur J Heart Fail, 2009, 11:223.
Sourgounis A, Lipiecki J, Lo TS, Hamon M.
Coronary stents and chronic anticoagulation.
Circulation 2009; 119:1682-1688.
Lambert JC, Schraen-Maschke S, Richard F, Fievet N, Rouaud O,
Berr C, Dartigues JF, Tzourio C, Alpérovitch A, Buée L, Amouyel P.
Association of plasma amyloid {beta} with risk of dementia : The
prospective Three-City Study.
Neurology, 2009, 73:847-853.
Sparks DL, Hunsaker JC 3rd, Amouyel P, Malafosse A, Bellivier F,
Leboyer M, Courtet P, Helbecque N.
Angiotensin I-converting enzyme I/D polymorphism and suicidal
behaviors.
Am J Med Genet B Neuropsychiatr Genet, 2009, 150B:290-294.
Lambert JC, Heath S, Even G, Campion D, Sleegers K, Hiltunen M,
Combarros O, Zelenika D, Bullido MJ, Tavernier B, Letenneur L,
Bettens K, Berr C, Pasquier F, Fiévet N, Barberger-Gateau P,
Engelborghs S, De Deyn P, Mateo I, Franck A, Helisalmi S, Porcellini E,
Hanon O; the European Alzheimer's Disease Initiative Investigators,
de Pancorbo MM, Lendon C, Dufouil C, Jaillard C, Leveillard T,
Alvarez V, Bosco P, Mancuso M, Panza F, Nacmias B, Bossù P,
Piccardi P, Annoni G, Seripa D, Galimberti D, Hannequin D, Licastro F,
Soininen H, Ritchie K, Blanché H, Dartigues JF, Tzourio C, Gut I,
Van Broeckhoven C, Alpérovitch A, Lathrop M, Amouyel P.
Genome-wide association study identifies variants at CLU and CR1
associated with Alzheimer's disease.
Nat Genet, 2009, 41:1094-1099.
Tilloy E, Montaye M, Kee F, Bingham A, Arveiler D, Ruidavets JB,
Evans A, Haas B, Ferrières J, Ducimetière P, Amouyel P, Dallongeville J.
Contribution of cardiovascular risk factors to coronary risk in patients with
intermittent claudication in the PRIME Cohort Study of European men.
Valgimigli M, Campo G, de Cesare N, Meliga E, Vranckx P, Furgieri A,
Angiolillo DJ, Sabatè M, Hamon M, Repetto A, Colangelo S,
Brugaletta S, Parrinello G, Percoco G, Ferrari R ; Tailoring Treatment
With Tirofiban in Patients Showing Resistance to Aspirin and/or
Resistance to Clopidogrel (3T/2R) Investigators.
Intensifying platelet inhibition with tirofiban in poor responders to
aspirin, clopidogrel, or both agents undergoing elective coronary
intervention: results from the double-blind, prospective, randomized
Tailoring Treatment with Tirofiban in Patients Showing Resistance to
Aspirin and/or Resistance to Clopidogrel study.
Circulation, 2009, 119:3215-3222.
Legry V, Bokor S, Cottel D, Beghin L, Catasta G, Nagy E, GonzalezGross M, Spinneker A, Stehle P, Molnar D, Moreno L, Amouyel P,
Dallongeville J, Meirhaeghe A.
Associations between common genetic polymorphisms in angiopoietinlike proteins 3 and 4 and lipid metabolism and adiposity in European
adolescents and adults.
J Clin Endoc Metab, 2009, 94:5070-5077.
Vereecken C, De Henauw S, Maes L, Moreno L, Manios Y, Phillipp K,
Plada M, De Bourdeaudhuij I ; HELENA Study Group.
Reliability and validity of a healthy diet determinants questionnaire for
adolescents.
Public Health Nutr, 2009, 12:1830-1838.
Legry V, Goumidi L, Huyvaert M, Cottel D, Ferrières J, Arveiler D,
Bingham A, Wagner A, Ruidavets JB, Ducimetière P, Amouyel P,
Meirhaeghe A.
Association study between angiopoietin-like 6 (ANGPTL6) gene
polymorphisms and Metabolic Syndrome-related phenotypes in the
French MONICA Study.
Diabetes Metab, 2009, 35:287-292.
Zerbib J, Seddon JM, Richard F, Reynolds R, Leveziel N, Benlian P,
Borel P, Feingold J, Munnich A, Soubrane G, Kaplan J, Rozet JM,
Souied EH.
rs5888 variant of SCARB1 gene is a possible susceptibility factor for agerelated macular degeneration.
PLoS One, 2009, 4:e7341.
Legry V, Cottel D, Ferrières J, Arveiler D, Andrieux N, Bingham A,
Wagner A, Ruidavets JB, Ducimetière P, Amouyel P, Meirhaeghe A.
Effect of a FTO polymorphism on fat mass, obesity and type 2 diabetes in
the French MONICA Study.
Metabolism, 2009, 58:971-975.
Meroufel D, Médiène-Benchekor S, Dumont J, Benhammamouch S,
Amouyel P, Brousseau T.
Relationship between endothelial nitric oxide synthase gene polymorphisms
and risk of myocardial infarction in the Algerian population.
Egypt J Med Hum Genetics, 2009, 10(1).
Lemesle G, Delhaye C, Sudre A, Broucqsault D, Rosey G, Bauters C,
Lablanche JM.
Impact of high loading and maintenance dose of clopidogrel within the
first 15 days after percutaneous coronary intervention on patient
outcome.
Am Heart J, 2009, 157:375-382.
Book Chapter
Hamon M, Mc Fadden E.
Trans-radial approach for cardiovascular interventions (2nd edition).
ESM Editions 2009.
Meroufel D, Dumont J, Médiène-Benchekor S, Benhammamouch S,
Ducimetière P, Cottel D, Montaye M, Amouyel P, Brousseau T.
Characterisation of arginase 1 gene polymorphisms in the Algerian
population and association with blood pressure.
Clin Biochem, 2009, 42:1178-1182.
2010
Agrinier N, Cournot M, Dallongeville J, Arveiler D, Ducimetière P,
Ruidavets JB, Ferrières J.
Menopause and modifiable coronary heart disease risk factors : A
population based study.
Maturitas, 2010, 65:237-243.
Mouquet F, Cuilleret F, Susen S, Sautière K, Marboeuf P, Ennezat PV,
Mc Fadden EP, Pigny P, Richard F, Hennache B, Vantyghem MC,
Dallongeville J, Jude B, Bertrand ME, Van Belle E.
Metabolic syndrome and collateral vessel formation in patients with
documented occluded coronary arteries : The impact of hyperglycemia,
insulin-resistance, adiponectin and PAI-1.
Eur Heart J, 2009, 30:840-849.
Bauters C, Ennezat PV, Lamblin N, de Groote P.
Left ventricular remodeling and heart failure after myocardial infarction
in elderly patients.
Am J Cardiol, 2010, 105:903-904.
118
Contents
Beseme O, Fertin M, Drobecq H, Amouyel P, Pinet F.
Combinatorial peptide ligand library plasma treatment : Advantages for
accessing low-abundance proteins.
Electrophoresis, 2010, 31:2697-2704.
de Groote P, Lamblin N, Launay D, Bervar JF, Hachulla E. [Screening
and diagnosis of pulmonary arterial hypertension.
Comments regarding the latest guidelines from the European Societies
of Cardiology and of Pulmonology].
Presse Med. 2010; 39 Suppl 1:1S16-21. [French].
Blacher J, Evans A, Arveiler D, Amouyel P, Ferrières J, Bingham A,
Yarnell J, Haas B, Montaye M, Ruidavets JB, Ducimetière P.
Residual cardiovascular risk in treated hypertension and hyperlipidaemia :
the PRIME Study.
J Hum Hypertens, 2010, 24:19-26.
Dubois E, Richard V, Mulder P, Lamblin N, Drobecq H, Henry JP,
Amouyel P, Thuillez C, Bauters C, Pinet F.
Decreased serine207 phosphorylation of troponin T as a biomarker for
left ventricular remodelling after myocardial infarction.
Eur Heart J, 2010, in press.
Bokor S, Legry V, Meirhaeghe A, Ruiz JR, Mauro B, Widhalm K,
Manios Y, Amouyel P, Moreno LA, Molnàr D, Dallongeville J and on
behalf of the HELENA Study group.
Single-nucleotide polymorphism of CD36 locus and obesity in European
adolescents.
Obesity, 2010, 18:1398-1403.
Dumont J, Jossé R, Lambert C, Anthérieu S, Le Hegarat L, Aninat C,
Robin M.A, Guguen-Guillouzo C, Guillouzo A.
Differential toxicity of heterocyclic aromatic amines and their mixture in
metabolically competent HepaRG cells.
Toxicol Appl Pharmacol, 2010, 245:256-263.
Bokor S, Meirhaeghe A, Ruiz JR, Zaccaria M, Widhalm K, Manios Y,
Gonzalez-Gross M, Amouyel P,. Moreno LA, Molnàr D, Dallongeville J,
on behalf of the HELENA Study group.
Common polymorphisms in 6 genes of methyl group metabolism
pathway and obesity in European adolescents.
Int J Pediatr Obes, 2010, in press.
Dupont A, Elkalioubie A, Juthier F, Tagzirt M, Vincentelli A,
Le Tourneau T, Haulon S, Deklunder G, Breyne J, Susen S,
Marechaux S, Pinet F, Jude B.
Frequency of abdominal aortic aneurysm in patients undergoing
coronary artery bypass grafting.
Am J Cardiol, 2010, 105:1545-1548.
Bokor S, Dumont J, Spinneker A, Gonzalez-Gross M, Nova E,
Widhalm K, Moschonis G, Stehle P, Amouyel P, De Henauw S,
Molnar D, Moreno LA, Meirhaeghe A, Dallongeville J.
Single nucleotide polymorphisms in the FADS gene cluster are associated
with delta-5 and delta-6 desaturase activities estimated by serum fatty
acid ratios.
J Lipid Res, 2010, 51:2325-2333.
Emerging Risk Factors Collaboration, Kaptoge S, Di Angelantonio E,
Lowe G, Pepys MB, Thompson SG, Collins R, Danesh J.
C-reactive protein concentration and risk of coronary heart disease,
stroke, and mortality : an individual participant meta-analysis.
Lancet, 2010, 375:132-140.
Emerging Risk Factors Collaboration, Sarwar N, Gao P, Seshasai SR,
Gobin R, Kaptoge S, Di Angelantonio E, Ingelsson E, Lawlor DA,
Selvin E, Stampfer M, Stehouwer CD, Lewington S, Pennells L,
Thompson A, Sattar N, White IR, Ray KK, Danesh J.
Diabetes mellitus, fasting blood glucose concentration, and risk of
vascular disease : a collaborative meta-analysis of 102 prospective studies.
Lancet, 2010, 375:2215-2222.
Canouï-Poitrine F, Luc G, Bard JM, Ferrières J, Yarnell J, Arveiler D,
Morange P, Kee F, Evans A, Amouyel P, Ducimetière P, Empana JP.
Relative contribution of lipids and apolipoproteins to incident Coronary
Heart Disease and Ischemic Stroke : The PRIME Study.
Cerebrovasc Dis, 2010, 30:252-259.
Cohen A, Dallongeville J, Durand-Zaleski I, Bouée S, Le Heuzey JY;
the EPHA Investigators.
Characteristics and management of outpatients with history of or current
atrial fibrillation : The observational French EPHA study.
Empana JP, Jouven X, Canouï-Poitrine F; Luc G, Tafflet M, Haas B,
Arveiler D, Ferrières J, Ruidavets JB; Montaye M, Yarnell J,
Morange P, Kee F, Evans A, Amouyel P, Ducimetière P.
C-Reactive Protein, Interleukin 6, Fibrinogen and risk of sudden death in
European middle-aged men : The PRIME Study.
Arterioscler Thromb Vasc Biol. 2010; 30(10):2047-2052.
Dallongeville J, Dauchet L, de Mouzon O, Réquillart V, Soler L-G.
Increasing fruit and vegetable consumption : a cost-effectiveness analysis
of public policies.
Eur J Public Health, 2010, in press.
Evangelou E, Maraganore DM, Annesi G, Brighina L, Brice A,
Elbaz A, Ferrarese C, Hadjigeorgiou GM, Krueger R, Lambert JC,
Lesage S, Markopoulou K, Mellick GD, Meeus B, Pedersen NL,
Quattrone A, Van Broeckhoven C, Sharma M, Silburn PA, Tan EK,
Wirdefeldt K, Ioannidis JP; for the Genetic Epidemiology of
Parkinson's Disease (GEOPD) Consortium Australia.
Non-replication of association for six polymorphisms from meta-analysis
of genome-wide association studies of Parkinson's disease : Large-scale
collaborative study.
Dallongeville J & Meirhaeghe A.
Triglyceride-mediated pathways and coronary heart disease.
Lancet, 2010, 376:956-957.
Dauchet L, Montaye M, Ruidavets JB, Arveiler D, Kee F, Bingham A,
Ferrières J, Haas B, Evans A, Ducimetière P, Amouyel P, Dallongeville J.
Association between the frequency of fruit and vegetable consumption
and cardiovascular disease in male smokers and non-smokers.
Eur J Clin Nutr, 2010, 64:578-586.
Fertin M, Beseme O, Duban S, Amouyel P, Bauters C, Pinet F.
Deep plasma proteomic analysis of patients with left ventricular
remodeling after a first myocardial infarction.
Proteomics Clin Appl, 2010, in press.
De Bacquer D, Dallongeville J, Kotseva K, Montaye M, Heidrich J,
Wood D, Keil U, De Backer G, EUROASPIRE Study Group.
The challenge facing the care of overweight coronary patients in Europe.
Eur J Cardiovasc Prev Rehabil, 2010, in press.
Gey KF, Ducimetière P, Evans A, Amouyel P, Arveiler D, Ferrières J,
Luc G, Kee F, Bingham A, Yarnell J, Cambien F.
Low plasma retinol predicts coronary events in healthy middle-aged men :
The PRIME Study.
De Bacquer D, Dallongeville J, Heidrich J, Kotseva K, Reiner Z,
Gaita D, Prugger C, Wellmann J, Wood D, De Backer G, Keil U,
on behalf of the EUROASPIRE III Study Group.
Management of overweight and obese patients with coronary heart
disease across Europe.
119
Contents
Goumidi L, Flamant F, Lendon C, Galimberti D, Pasquier F, Scarpini E,
Hannequin D, Campion D, Amouyel P, Lambert JC, Meirhaeghe A.
Study of thyroid hormone receptor alpha gene polymorphisms on
Neurobiol Aging, 2010, in press.
Lambert JC, Sleegers K, González-Pérez A, Ingelsson M,
Beecham GW, Hiltunen M, Combarros O, Bullido MJ, Brouwers N,
Bettens K, Berr C, Pasquier F, Richard F, Dekosky ST, Hannequin D,
Haines JL, Tognoni G, Fiévet N, Dartigues JF, Tzourio C, Engelborghs S,
Arosio B, Coto E, De Deyn P, Del Zompo M, Mateo I, Boada M,
Antunez C, Lopez-Arrieta J, Epelbaum J, Schjeide BM, FrankGarcia A, Giedraitis V, Helisalmi S, Porcellini E, Pilotto A, Forti P,
Ferri R, Delepine M, Zelenika D, Lathrop M, Scarpini E, Siciliano G,
Solfrizzi V, Sorbi S, Spalletta G, Ravaglia G, Valdivieso F, Vepsäläinen S,
Alvarez V, Bosco P, Mancuso M, Panza F, Nacmias B, Bossù P,
Hanon O, Piccardi P, Annoni G, Mann D, Marambaud P, Seripa D,
Galimberti D, Tanzi RE, Bertram L, Lendon C, Lannfelt L, Licastro F,
Campion D, Pericak-Vance MA, Soininen H, Van Broeckhoven C,
Alpérovitch A, Ruiz A, Kamboh MI, Amouyel P.
The CALHM1 P86L polymorphism is a genetic modifier of age at onset in
Alzheimer's Disease: a meta-analysis study.
J Alzheimers Dis, 2010, in press.
Gruson E, Montaye M, Kee F, Wagner A, Bingham A, Ruidavets JB,
Haas B, Evans A, Ferrières J, Ducimetière P, Amouyel P, Dallongeville J.
Anthropometric assessment of abdominal obesity and coronary heart
disease risk in men : the PRIME study.
Heart, 2010, 96:136-140.
Hachulla E, Launay D, Yaici A, Berezne A, de Groote P, Sitbon O,
Mouthon L, Guillevin L, Hatron PY, Simonneau G, Clerson P,
Humbert M; French PAH-SSc Network.
Pulmonary arterial hypertension associated with systemic sclerosis in
patients with functional class II dyspnoea : mild symptoms but severe
outcome.
Lamblin N, Ratajczak P, Hot D, Dubois E, Chwastyniak M, Beseme O,
Drobecq H, Lemoine Y, Koussa M, Pinet F, Amouyel P.
Profile of macrophages in human abdominal aortic aneurysms : a
transcriptomic, proteomic and antibody protein array study.
J Proteome Res, 2010, 9:3720-3729.
Hansmannel F, Sillaire A, Kamboh MI, Lendon C, Pasquier F,
Hannequin D, Laumet G, Mounier A, Ayral A-M, Dekosky ST,
Hauw J-J, Berr C, Mann D, Amouyel P, Campion D, Lambert JC.
Is the urea cycle involved in Alzheimer's disease ?
J Alzheimers Dis, 2010, 21:1013-1021.
Laumet G, Petitprez V, Sillaire A, Ayral AM, Hansmannel F, Chapuis J,
Hannequin D, Pasquier F, Scarpini E, Galimberti D, Lendon C,
Campion D, Amouyel P, Lambert JC.
A study of the association between the ADAM12 and SH3PXD2A
(SH3MD1) genes and Alzheimer's disease.
Neurosci Lett, 2010, 468:1-2.
Hong MG, Alexeyenko A, Lambert JC, Amouyel P, Prince JA.
Genome-wide pathway analysis implicates intracellular transmembrane
protein transport in Alzheimer disease.
J Hum Genet, 2010, in press.
Krüger R, Sharma M, Riess O, Gasser T, Van Broeckhoven C,
Theuns J, Aasly J, Annesi G, Bentivoglio AR, Brice A, Djarmati A,
Elbaz A, Farrer M, Ferrarese C, Gibson JM, Hadjigeorgiou GM,
Hattori N, Ioannidis JP, Jasinska-Myga B, Klein C, Lambert JC,
Lesage S, Lin JJ, Lynch T, Mellick GD, de Nigris F, Opala G,
Prigione A, Quattrone A, Ross OA, Satake W, Silburn PA, Tan EK,
Toda T, Tomiyama H, Wirdefeldt K, Wszolek Z, Xiromerisiou G,
Maraganore DM ; for the Genetic Epidemiology of Parkinson's
disease consortium.
A large-scale genetic association study to evaluate the contribution of
Omi/HtrA2 (PARK13) to Parkinson's disease.
Laumet G, Chouraki V, Grenier Boley B, Legry V, Heath S, Zelenika D,
Fievet N, Hannequin D, Delepine M, Pasquier F, Hanon O, Brice A,
Epelbaum J, Berr C, Dartigues J-F, Tzourio C, Campion D, Lathrop M,
Bertram L, Amouyel P, Lambert JC.
Systematic analysis of candidate genes for Alzheimer’s disease in a
French, genome-wide association study.
J Alzheimers Dis, 2010, 20:1181-1188.
Le Fur I, Laumet G, Richard F, Fievet N, Berr C, Rouaud O, Delcourt C,
Association study of the CFH Y402H polymorphism with Alzheimer's
disease.
Labayen I, Ruiz JR, Meirhaeghe A, Ortega FB, Jiménez-Pavón D,
Castillo MJ, De Henauw S, González-Gross M, Bueno G, Molnar D,
Kafatos A, Esperanza L, Moreno LA.
Association between the FTO rs9939609 polymorphism and leptin in
European adolescents : a possible link with energy balance control. The
HELENA study.
Int J Obes, 2010, in press.
Lemesle G, Sudre A, Bouallal R, Delhaye C, Rosey G, Bauters C,
Lablanche JM.
Impact of thrombus aspiration use and direct stenting on final
myocardial blush score in patients presenting with ST-elevation
myocardial infarction.
Deciphering genetic susceptibility to frontotemporal lobar dementia.
Nat Genet, 2010, 42:189-190.
Lemesle G, Maluenda G, Bonello L, Delhaye C, Sudre A, Bauters C,
Lablanche JM.
Dosing strategies for antiplatelet therapy in percutaneous coronary
intervention.
Hosp Pract (Minneap), 2010, 38:50-58.
Lambert JC, Grenier-Boley B, Chouraki V, Heath S, Zelenika D,
Fievet N, Hannequin D, Pasquier F, Hanon O, Brice A, Epelbaum J,
Berr C, Dartigues J-F, Tzourio C, Campion D, Lathrop M, Amouyel P.
Implication of the immune system in Alzheimer’s disease : evidence from
genome-wide pathway analysis.
J Alzheimers Dis, 2010, 20:1107-1118.
Le Tourneau T, Betto M, Richardson M, Juthier F, Ennezat PV,
Polge AS, Bauters C, Vincentelli A, Deklunder G.
Prospective assessment of multiple cardiac papillary fibroelastomas An
echocardiographic and surgical study.
Int J Cardiol, 2010, in press.
Le Tourneau T, Richardson M, Juthier F, Modine T, Fayad G, Polge AS,
Ennezat PV, Bauters C, Vincentelli A, Deklunder G.
Echocardiography predictors and prognostic value of pulmonary artery
systolic pressure in chronic organic mitral regurgitation.
Heart, 2010, 96:1311-1317.
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Leveziel N, Puche N, Richard F, Somner JE, Zerbib J, Bastuji-Garin S,
Cohen S, Korobelnik JF, Sahel JA, Soubrane G, Benlian P, Souied EH.
Genotypic influences on severity of exudative Age-related Macular
Degeneration.
Invest Ophthalmol Vis Sci, 2010, 51:2620-2625.
Sleegers K, Lambert JC, Bertram L, Cruts M, Amouyel P,
Van Broeckhoven C.
The pursuit of susceptibility genes for Alzheimer's disease : progress and
prospects.
Trends Genet, 2010, 26:84-93.
Luc G, Empana JP, Morange P, Juhan-Vague I, Arveiler D, Ferrieres J,
Amouyel P, Evans A, Kee F, Bingham A, Machez E, Ducimetiere P.
Adipocytokines and the risk of coronary heart disease in healthy middle
aged men : the PRIME Study.
Int J Obes (Lond). 2010; 34(1):118-126.
Straczek C, Ducimetiere P, Barberger-Gateau P, Helmer C, Ritchie K,
Jouven X, Carcaillon L, Amouyel P, Tzourio C, Empana JP.
Higher level of systemic C-Reactive Protein is independently predictive of
Coronary Heart Disease in Older Community-Dwelling Adults: the ThreeCity study.
J Am Geriatr Soc, 2010, 58:129-35.
Mekinian A, Lions C, Leleu X, Duhamel A, Lamblin N, Coiteux V,
De Groote P, Hatron PY, Facon T, Beregi JP, Hachulla E, Launay D ;
Lille Amyloidosis Study Group.
Prognosis assessment of cardiac involvement in systemic AL amyloidosis
by magnetic resonance imaging.
Am J Med, 2010, 123:864-868.
Szopa M, Meirhaeghe A, Luan J, Moreno LA, Gonzalez-Gross M,
Vidal-Puig A, Cooper C, Hagen R, Amouyel P, Wareham NJ, Loos RJF.
No association between polymorphisms in the INSIG1 gene and the risk
of type 2 diabetes and related traits.
Am J Clin Nutr, 2010, 92:252-257.
Moliner-Urdiales D, Ruiz JR, Vicente-Rodriguez G, Ortega FB,
Rey-Lopez JP, España-Romero V, Casajús JA, Molnar D, Widhalm K,
Dallongeville J, González-Gross M, Castillo MJ, Sjöström M,
Moreno LA.
Associations of muscular and cardiorespiratory fitness with total and
central body fat in adolescents ; The HELENA Study.
Br J Sports Med, 2010, in press.
Tilloy E, Cottel D, Ruidavets JB, Arveiler D, Ducimetière P,
Bongard V, Haas B, Ferrières J, Wagner A, Bingham A, Amouyel P,
Dallongeville J.
Characteristics of current smokers, former smokers and secondhand
exposure and evolution between 1985 and 2007.
Eur J Cardiovasc Prev Rehabil. 2010, in press.
Triglyceride Coronary Disease Genetics Consortium and Emerging
Risk Factors Collaboration, Sarwar N, Sandhu MS, Ricketts SL,
Butterworth AS, Di Angelantonio E, Boekholdt SM, Ouwehand W,
Watkins H, Samani NJ, Saleheen D, Lawlor D, Reilly MP, Hingorani
AD, Talmud PJ, Danesh J.
Triglyceride-mediated pathways and coronary disease: collaborative
analysis of 101 studies.
Lancet, 2010, 375:1634-1639.
Pigeyre M, Bokor S, Romon M, Gottrand F, Gilbert CC, Valtueña J,
Gómez-Martínez S, Moreno LA, Amouyel P, Dallongeville J,
Meirhaeghe A.
Influence of maternal educational level on the association between the
rs3809508 neuromedin B gene polymorphism and the risk of obesity in
the HELENA study.
Int J Obes (Lond), 2010, 3:478-486.
Plichart M, Barberger-Gateau P, Tzourio C, Amouyel P, Pérès K,
Ritchie K, Jouven X, Ducimetière P, Empana JP.
Disability and incident coronary heart disease in older communitydwelling adults: the Three-City Study.
J Am Geriatr Soc, 2010, 58:636-642.
Van Belle E, Dallongeville J, Vicaut E, Degrandsart A, Baulac C,
Montalescot G, on behalf of The OPERA Investigators.
Ischaemia-modified albumin levels predict long-term outcome in
patients with acute myocardial infarction. The French Nationwide OPERA
study.
Am Heart J, 2010, 159:570-576.
Rodriguez-Artalejo F, Guallar E, Borghi C, Dallongeville J,
De Backer G, Halcox JP, Hernandez-Vecino R, Jimenez FJ, MassoGonzalez EL, Perk J, Steg PG, Banegas JR, Investigators E.
Rationale and methods of the European Study on Cardiovascular Risk
Prevention and Management in Daily Practice (EURIKA).
BMC Public Health, 2010, 10:382.
Vasseur F, Caeyseele T, Barat-Houari M, Lobbens S, Meirhaeghe A,
Meyre D, Froguel P, Amouyel P, Helbecque N.
Concordance of two multiple analytical approaches demonstrate that
interaction between BMI and ADIPOQ haplotypes is a determinant of LDL
cholesterol in a general French population.
J Hum Genet, 2010, 55:227-231.
Ruiz JR, Labayen I, Ortega FB, Legry V, Moreno LA, Dallongeville J,
Martínez-Gómez D, Bokor S, Manios Y, Ciarapica D, Gottrand F,
De Henauw S, Molnar D, Sjöström M, Meirhaeghe A.
Physical activity attenuates the effect of the FTO rs9939609
polymorphism on total and central body fat in adolescents ; The HELENA
Study.
Arch Pediatr Adolesc Med. 2010; 164(4):328-333.
Verier C, Meirhaeghe A, Bokor S, Breidenassel C, Manios, Molnár D,
Artero EG, Nova E, De Henauw S, Moreno LA, Amouyel P, Labayen I,
Bevilacqua N, Turck D, Béghin L, Dallongeville J, Gottrand F.
Breastfeeding modulates the influence of the peroxisome proliferatoractivated receptor gamma (PPARG) Pro12Ala polymorphism on adiposity
in adolescents : the HELENA Cross-Sectional Study.
Diabetes Care, 2010, 33:190-196.
Seshadri S, Fitzpatrick AL, Ikram A, DeStefano AL, Gudnason V,
Boada M, Bis JC, Smith AV, Carassquillo MM, Lambert JC, Harold D,
Schrijvers EMC, Ramirez-Lorca R, Debette S, Longstreth WT,
Janssens CJW,Pankratz, Dartigues J-F, Hollinworth P, Aspelund T,
Hernadez I, Beiser A, Kuller LH, Koudstaal PJ, Dickson DW,
Tzourio C, Abraham R, Antunez C, Du, Y, Rotter JI, Aulchenko YS,
Harris TB, Petersen RC, Berr C, Owen MJ, Lopez-Arrieta J,
Varadarajan BN, Becker JT, Rivadeneira F, Nalls MA, GraffRadford NR, Campion D, Auerbach S, Rice K, Hofman A, Jonsson PV,
Schmidt H, Lathrop M, Mosley TH, Au R, Psaty BM, Uitterlinden AG,
Farrer LA, Lumley T, Ruiz A, Williams J, Amouyel P, Younkin SG,
Wolf PA, Launer LJ, Lopez O, Van Duijn C, Breteler M on behalf of
the Charge, GERAD1 and EADI1 consortia.
Genome-wide association studies of alzheimer’s disease.
JAMA, 2010, 303:1864-65.
Wagner A, Sadoun A, Dallongeville J, Ferrières J, Amouyel P,
Ruidavets JB, Arveiler D.
High blood pressure prevalence and control in a middle-aged French
population and their associated factors : the MONA LISA study.
J Hypertens, 2010, in press.
Zerbib J, Richard F, Puche N, Leveziel N, Cohen SY, Korobelnik JF,
Sahel J, Munnich A, Kaplan J, Rozet JM, Souied EH.
R102G polymorphism of the C3 gene associated with exudative agerelated macular degeneration in a French population.
Mol Vis, 2010, 16:1324-1330.
121
Contents
Jiménez Pavón D, Ortega FP, Ruiz JR, España Romero V,
García Artero E, Moliner Urdiales D, Gómez Martínez S,
Vicente Rodríguez G, Manios Y, Béghin L, Répasy J, Sjöstrom M,
Moreno LA, González Gross M, Castillo MJ; HELENA Study Group.
Socioeconomic status influences physical fitness in European adolescents
independently of body fat and physical activity : the HELENA study.
Nutr Hosp, 2010, 25:311-316.
Rey-López JP, Vicente-Rodriguez G, Ortega FB, Ruiz JR, MartinezGómez D, De Henauw S, Manios Y, Molnar D, Polito A, Verloigne M,
Castillo MJ, Sjöström M, De Bourdeaudhuij I, Moreno LA ; HELENA
Study Group.
Sedentary patterns and media availability in European adolescents : The
HELENA study.
Prev Med, 2010, 51:50-55.
PhD
Amélie BRUANDET
Directeur de thèse : Philippe Amouyel
« Facteurs pronostiques de patients atteints de démence suivis en centre
mémoire de ressource et de recherche : exemple d'utilisation de bases de
données médicales à des fins de recherche clinique »
23 septembre 2008
Vanessa LEGRY
Directeur de thèse : Aline Meirhaeghe-Hurez
« Recherche de déterminants génétiques des phénotypes associés au
syndrome métabolique en population »
28 septembre 2009
122
Contents
abdominal aortic aneurysms and chronic heart failure. These two
affections are subject to remodelling mechanisms affecting the
cardiovascular system, whose implication in the prognosis of the
patients is critical. Indeed, in consideration of the improved
treatment of the coronary disease and increased life expectancy
of patients, the structural modifications of the cardiovascular
system are now prevailing affecting the prognosis and quality
of life of individuals.
Search for the molecular
determinants of cardiovascular diseases via
proteomic analysis
and candidate gene
approaches
The strategy used consists in comparing the proteome of cells
in patients suffering from these affections to that of cells in nonaffected subjects. Cells used for these comparisons are
macrophages and smooth muscle cells. Bi-dimensional
electrophoresis and protein chips techniques are used.
Significant efforts made to analyse these data using biocomputing before and after these experiments are made. This
approach has already been validated and maps for the
proteome of macrophages (Dupont et al, 2004) and smooth
muscle cells (Dupont et al, 2005), which are critical for the
interpretation of data, were produced by the unit. Our team is
also developing proteomic techniques to increase sensitivity
and be able to analyse proteomes on reduced quantities of
tissues, or even directly on plasma or serum to adjust to the
necessarily limited biological resources of clinical studies in
humans. These experiments have therefore enabled us to
validate the 2D-DIGE Saturation Dye technique applied to
samples collected as part of clinical protocols (Dupont et al,
2008). However, certain proteins having a low molecular weight
(<20 kDa) or hydrophobic proteins that included membrane
proteins cannot be detected by 2-D electrophoresis techniques.
This is why we turned to the use of protein biochips. These
experiments have enabled us to validate the SELDI-TOF
technique to identify differentially expressed proteins having a
low molecular weight. This technique comes in addition to 2D
electrophoresis (Cieniewski-Bernard et al, 2008).
Florence PINET
DR 2 Inserm
Group Members :
Bauters Christophe, PU PH Lille 2
Degroote Pascal, PU CHRU Lille
Hamon Martial, PU-PH CHU Caen
Huygens Maggy, Technician IPL
Beseme Olivia, Engineer Lille 2
Acosta Adélina, Postdoctoral fellow
Dubois Emilie, Postdoctoral fellow
Boytard Ludovic, PhD Student Lille 2
Fertin Marie, PhD Student Lille 2
Burdese Justine, Master 2 Paris VII
Key words :
Differential proteomics. Bioinformatics. Biomarkers. Cardiovascular diseases (coronary heart disease, aneurysm, heart
failure). Risk factors. Translational research.
In the past thirty years, more than 200 cardiovascular disease
risk factors have been identified including approximately ten
modifiable risk factors - which seem to account for the major
part of avoidable cases in a population. However, considering
their interactions with each other, their impact on the
individual risk varies significantly. The determinants for these
variations are difficult to assess and are often related to
individual susceptibilities.
These various techniques are then applied to clinical studies
developed with cardiologists. These studies have been
designed to maximise differences between cases and. So, the
LILAS protocol is a case-control study which included 24 men
suffering from abdominal aortic aneurysms of atherosclerotic
origin first treated using surgery and 21 men affected by
obliterating peripheral arteriopathy treated by surgery
though with no abdominal aortic aneurysms. For each
subject, we preserve serum, and isolate macrophages and
smooth muscle cells with an explant of a fragment of the
vascular segment obtained after surgical excision. We have
also finalised the REVE study whose primary objective is to
identify new determinants for post-infarction left ventricular
remodelling. Patients included had to have suffered from an
inaugural myocardial infarction of anterior topography; they
were regularly examined using cardiac echography – the first
time 7 days after, the 2nd time 3 months after and the last
time during examinations performed after 1 year. REVE-1
findings showed that, unfortunately, left ventricular
remodelling still remained a relatively frequent development
(affecting one third of all patients followed-up in the REVE-1
study) (Savoye et al, 2006). In the REVE 1 study, we developed
experimental conditions on the basis of 4 control samples
and 4 patients’ samples on 3 types of chips - hydrophobic
(H50), anionic (Q10) and cationic (CM10) chips. The sample of
REVE (n=93) was divided into tertiles and we compared
samples from tertile 1 (n=31) (non-remodelling patients) with
These individual susceptibilities are analysed using candidate
gene approaches. This is how, for heart failure - a condition
whose prognosis still is severe - we have studied the association
of various functional polymorphisms of physiopathological
pathways involved in its development or prognosis. We were
therefore able to show, in a clinical series of more than 1,200
patients suffering from heart failure (one of the largest cohorts
on the genetics of heart failure developed with the cardiologists
of the CHRU of Lille), the prognosis role of polymorphisms in
metalloproteases 3 and 9 genes (Mizon-Gérard et al, 2004) and
the absence of any association with the polymorphisms of the
genes coding beta-adrenergic receptors (de Groote et al, 2005)
or AMDP1 (de Groote et al, 2007). In addition to genetic
polymorphisms, we have also shown the relevance of two
biochemical markers (Lamblin et al, 2005a & 2005b).
In order to identify further markers for these risk variations, we
have developed a strategy for the search of original susceptibility
genes on the basis of differential proteomic analyses. Our close
cooperation with cardiologists and their active involvement in
research efforts conducted in the laboratory have enabled us to
set up clinical studies to collect biological samples adapted to
these approaches. Two specific pathological fields were selected:
123
Contents
Bauters C, Ennezat PV, Tricot O, Lauwerier B, Lallemant R,
Saadouni H, Quandalle P, Jaboureck O, Lamblin N, Le Tourneau T ;
on behalf of The REVE Investigators.
Stress hyperglycaemia is an independent predictor of left ventricular
remodeling after first anterior myocardial infarction in non-diabetic patients.
Eur Heart J, 2007, 28:546-552.
Bauters A, Ennezat PV, Tricot O, Lallemant R, Aumegeat V,
Segrestin B, Quandalle P, Lamblin N, Bauters C ; REVE Investigators.
Relation of admission white blood cell count to left ventricular
remodeling after anterior wall acute myocardial infarction.
Am J Cardiol, 2007, 100:182-184.
samples from tertile 3 (n=31) (patients having left ventricular
remodelling exceeding 20%). We have found 4 differentially
expressed peaks. We have developed elution conditions for
each peak to be able to identify them using MALDI-TOF and
LC/MS-MS. Three peaks correspond to one protein and one
to another. We confirmed the identification of peaks through
the immuno-depletion of patients’ plasma using specific
antibodies. We were able to evidence that differences in the
expression of all 3 peaks corresponded to post-translational
variants of chain α1 of haptoglobin and the other to
hemoglobin (Pinet et al, 2008).
Brasselet C, Tassan S, Nazeyrollas P, Hamon M, Metz D.
Randomized comparison of femoral versus radial approach for
percutaneous coronary intervention using abxicimab in acute
myocardial infarction : Results of the FARMI (Five french Arterial access
with Reopro in Myocardial Infarction) Trial.
Heart, 2007, 93:1556-1561.
Cutlip DE, Windecker S, Mehran R, Boam A, Cohen DJ,
van Es G-A, Steg PG, Morel M-A, Mauri L, Vranckx P, McFadden E,
Lansky A, Hamon M, Krucoff MW, Serruys PW.
Clinical endpoints in coronary stent trials. A case for standardized definitions.
Circulation, 2007, 115:2344-2351.
Finally, in order to limit variability and to have more protein
material, we applied our techniques to an animal model
(LREVMODEL study). This is a heart failure experimental model
obtained in rats by ligaturing the coronary artery, developed in
cooperation with Dr V Richard, Dr P Mulder and Pr C Thuillez at
INSERM in Rouen, France. The first part of the differential
proteomic analysis was made on blood and heart tissue samples
2 months after ligaturing. Concerning the analysis of the left
ventricle, using 2D electrophoresis, we highlighted 49
differentially expressed polypeptidic spots between the left
ventricle (LV) of HF rats and controls. We have identified 27
different proteins involved in oxidative stress and proteasomal
degradation pathways, energetic metabolism with, among other
things, an increased expression of glycolytic enzymes and
reduced expression of proteins involved in the degradation
pathway of fatty acids or molecular chaperones, including HSP’s,
as well as proteins involved in the kinin pathway. This was how
we were able to show that proteins involved in oxidative stress
were modulated differently in the early and late phases of left
ventricular remodelling and differently depending on the severity of heart failure (Cieniewski-Bernard et al, review underway).
Debette S, Bauters C, Leys D, Lamblin N, Pasquier F, de Groote P.
Prevalence and determinants of cognitive impairment in chronic heart
failure patients.
Congest Heart Fail, 2007, 13(4):205-208.
de Groote P, Mouquet F, Dallongeville J, Lamblin N, Bauters C.
The impact of the AMPD1 gene polymorphism on exercise capacity, other
prognostic parameters, and survival in patients with stable congestive
heart failure. A study on 686 consecutive patients.
Am Heart J, 2007, 153:e15. [Letter to the editor.]
de Groote P, Delour P, Mouquet F, Lamblin N, Dagorn J, Hennebert O,
Le Tourneau T, Foucher-Hossein C, Verkindère C, Bauters C.
The effects of beta-blockers in patients with stable chronic heart failure.
Predictors of left ventricular ejection fraction improvement and impact on
prognosis.
Am Heart J, 2007, 154:589-595.
de Groote P, Ennezat PV, Mouquet F.
Bisoprolol in the treatment of chronic heart failure.
Vasc Health Risk Manag, 2007, 3:431-439.
After these new known or unknown proteins were determined,
we tried to go back to the gene and search for polymorphisms.
The new potential genetic susceptibility factors so identified
were then tested in the epidemiological studies defined in team
1 on the vascular risk and in studies used by team 3 on the
neurodegenerative risk. Transcriptome analyses using tailormade chips were also carried out and a DNA chip was made to
incorporate known genes suspected of being involved in
remodelling (family of metalloproteases).
de Groote P, Isnard R, Assyag P, Clersson P, Ducardonnet A,
Galinier M, Jondeau G, Leurs I, Thébaut JF, Komajda M.
Is the gap between guidelines and clinical practice in heart failure
treatment being filled ? Insights from the IPACT RECO survey.
Eur J Heart Fail, 2007, 9:1205-1211.
Dumont J, Zureik M, Bauters C, Grupposo MC, Cottel D, Montaye M,
Hamon M, Ducimetière P, Amouyel P, Brousseau T.
Association of OAZ1 Gene Polymorphisms With Subclinical and Clinical
Vascular Events.
Publications
Ennezat PV, Gonin X, Darchis J, de Groote P, Lamblin N, Bauters C,
Lejemtel T, Asseman P.
Delayed recovery of left ventricular systolic dysfunction : ''give time to
medical therapy''.
Minerva Cardioangiol, 2007, 55:426-427.
2007
Bauters C, Lamblin N, Ennezat PV, Mycinski C, Tricot O, Nugue O,
Segrestin B, Hannebicque G, Agraou B, Polge AS, de Groote P,
Helbecque N, Amouyel P; REVE Investigators.
A prospective evaluation of left ventricular remodeling after inaugural
anterior myocardial infarction as a function of gene polymorphisms in the
renin-angiotensin-aldosterone, adrenergic, and metalloproteinase systems.
Am Heart J. 2007; 153:641-648.
Hamon M, Gomes S, Clergeau M-R, Fradin S, Morello R, Hamon M.
Risk of Acute Brain Injury Related to Cerebral Microembolism During
Cardiac Catheterization Performed by Right Upper Limb Arterial Access.
Stroke, 2007, 2176-2179.
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Contents
Hamon M, Burzotta F, Oppenheim C, Morello R, Viader F, Hamon M.
Silent Cerebral Infarct After Cardiac Catheterization as Detected by
Diffusion Weighted Magnetic Resonnance Imaging : A Randomized
Comparison of Radial and Femoral arterial approaches.
Trials, 2007, 8:15.
Valgimigli M, Bolognese L, Anselmi M, Campo G, Rodriguez AE, de
Cesare N, Cohen DJ, Sheiban I, Colangelo S, Pasquetto G, Hamon M,
Vranckx P, Ferrario M, Prati F, Agostoni P, Malagutti P, Arcozzi C,
Parrinello G, Vassanelli C, Ferrari R, Percoco G.
Two-by-two factorial comparison of high-bolus-dose tirofiban followed by
standard infusion versus abciximab and sirolimus-eluting versus baremetal stent implantation in patients with acute myocardial infarction:
Design and rationale for the MULTI-STRATEGY trial.
Am Heart J, 2007, 154:39-45.
Hamon M, Morello R, Riddell JW, Hamon M.
Coronary arteries : Diagnostic performance of 16- versus 64-slice spiral
computed tomography of coronary arteries as compared to invasive
coronary angiography : Meta-analysis.
Radiology, 2007, 245:720-731.
de Groote P.
Cardiologies et maladies vasculaires. Indications thérapeutiques.
Masson, Paris, 2007, pages 719-722.
Komajda M, Amouyel P, Johnson N, Bergougnoux L, Laperche T,
de Groote P, Jaillon P, Cohen-Solal A ; Comité scientifique d'étude
de KEOPS et des investigateurs.
Treating heart failure with carvedilol in private practice (initiating
treatment and follow-up at one year). The KEOPS study.
Arch Mal Coeur Vaiss, 2007, 100:818-826.
de Groote P.
Cardiologies et maladies vasculaires. Cœurs et maladies systémiques
auto-immunes.
Masson, Paris, 2007, pages 1209-1223.
Launay D, Lambert M, Hachulla E, de Groote P, Remy-Jardin M,
Queyrel V, Morell-Dubois S, Charlanne H, Cortot A, Hatron PY.
Entéropathie exsudative révélant une péricardite chronique constrictive
idiopathique.
Rev Med Interne, 2007, 28:38-41.
de Groote P.
Traité de médecine cardiovasculaire du sujet âgé. Utilisation des dérivés
nitrés et médicaments apparentés.
Médecine-Sciences, Flammarion, Paris 2007, pages 510-514.
de Groote P.
Cardiologie et pathologies vasculaires. L’insuffisance cardiaque.
Med-Line Editions, Paris 2007, pages 261-284.
Le Hello C, Morello R, Lequerrec A, Duarte C, Riddell J, Hamon M.
Effect of PlA1/A2 glycoprotein IIIa gene polymorphism on the long-term
outcome after successful coronary stenting.
Thrombosis Journal, 2007, 5:19.
Dupont A, Pinet F.
The proteome and secretome of human arterial smooth muscle cell.
Methods Mol Biol, 2007, 357:225-233.
Manoukian SV, Feit F, Mehran R, Voeltz MD, Ebrahimi R, Hamon M,
Dangas GD, Lincoff AM, White HD, Moses JW, King SB 3rd,
Ohman EM, Stone GW.
Impact of major bleeding on 30-day mortality and clinical outcomes in
patients with acute coronary syndromes : an analysis from the ACUITY Trial.
J Am Coll Cardiol, 2007, 49:1362-1368.
Pinet F.
Isolation and analysis of renal vessels.
In : Renal and urinary Proteomics, Ed. Thongboonkerd, Wiley. 2007.
2008
Maréchaux S, Ennezat PV, Lejemtel TH, Polge AS, de Groote P,
Asseman P, Nevière R, Le Tourneau T, Deklunder G.
Left ventricular response to exercise in aortic stenosis: an exercise
echocardiographic study.
Echocardiography, 2007, 24:955-959.
Agostini D, Verberne HJ, Hamon M, Jacobson AF, Manrique A.
Cardiac (123)I-MIBG scintigraphy in heart failure.
Q J Nucl Med Mol Imaging, 2008, 52:369-377.
Bauters C.
Silent coronary plaque rupture.
Marquié C, Duchemin A, Klug D, Lamblin N, Mizon F, Cordova H,
Boulo M, Lacroix D, Pol A, Kacet S.
Can we implant cardioverter defibrillator under minimal sedation ?
Europace 2007, 9:545-550.
Bauters C.
Pathophysiology of coronary artery disease.
Rev Prat, 2008, 58:1523-1526.
Meirhaeghe A, Sandhu MS, McCarthy MI, de Groote P, Cottel D,
Arveiler D, Ferrières J, Groves CJ, Hattersley AT, Hitman GA,
Walker M, Wareham NJ, Amouyel P.
Association between the T-381C polymorphism of the brain natriuretic
peptide gene and risk of type 2 diabetes in human populations.
Hum Mol Genet, 2007, 16:1343-1350.
Bauters C, Lamblin N, de Groote P.
High-sensitivity C-reactive protein for risk stratification in patients with
heart failure.
Am Heart J, 2008 Feb, 155:e7.
Riddell JW, Chiche L, Plaud B, Hamon M.
Coronary stents and noncardiac surgery.
Circulation, 2007, 116:e378-e382.
Burzotta F, Trani C, Hamon M, Amoroso G, Kiemeneij F.
Transradial approach for coronary angiography and interventions in
patients with bypass grafts : tips and tricks.
Cath Cardiovasc Interv, 2008, 72: 263-72.
Schmidt J, Launay D, Soudan B, Hachulla E, de Groote P, Lambert M,
Queyrel V, Morell-Dubois S, Hatron PY.
Intérêt du dosage plasmatique de l’endothéline au cours de la
sclérodermie systémique.
Rev Med Interne, 2007, 28:371-376.
Cieniewski-Bernard C, Mulder P, Henry JP, Drobecq H, Dubois E,
Pottiez G, Thuillez C, Amouyel P, Richard V, Pinet F.
Proteomic analysis of left ventricular remodelling in an experimental
model of heart failure.
J Proteome Res, 2008, 7:5004-5016.
Cieniewski-Bernard C, Acosta A, Dubois E, Lamblin N, Beseme O,
Chwastyniak M, Amouyel P, Bauters C, Pinet F.
Proteomic analysis in cardiovascular diseases.
Clin Exp Pharmacol Physiol, 2008, 35:362-366.
125
Contents
Clergeau MR, Morello R, Le Page O, Hamon M.
Intérêt des statines dans la prévention des infarctus post-angioplastie
coronaire. Revue systématique et méta-analyse.
Ann Cardiol Ang, 2008, 57:181-186.
Hamon M, Baron J-C, Viader F, Hamon M.
Periprocedural stroke and cardiac catheterization.
Circulation, 2008, 118: 678-683.
Hamon M, Champ-Rigot L, Morello R, Riddell JW, Hamon M.
Diagnostic accuracy of in-stent coronary restenosis detection with
multislice spiral computed tomography: a meta-analysis.
Eur Radiol, 2008, 18:217-225.
Coutance G, Le Page O, Lo T, Hamon M.
Prognostic value of Brain Natriuretic Peptide in acute pulmonary embolism.
Critical Care, 2008, 12:R109.
de Groote P, Herpin D, Diévart F, Hanon O, Trochu JN, Artigou JY,
Galinier M, Habib G, Juillière Y, Neuder Y, Roudaut R, Komajda M.
Treatment of heart failure with preserved systolic function.
Hamon M, Lepage O, Malagutti P, Riddell JW, Morello R, Agostini D,
Hamon M.
Diagnostic performance of 16- and 64-section spiral CT for coronary
artery bypass graft assessment : Meta-analysis.
Radiology, 2008, 247:679-686.
de Groote P, Lamblin N, Mouquet F, Bauters C.
No gender survival difference in a population of patients with chronic heart
failure related to left ventricular systolic dysfunction and receiving optimal
medical therapy.
Hamon M, Nolan J.
Should radial artery access be the gold standard for PCI ?
Heart, 2008, 94:1530-1532.
Lemesle G, Sudre A, Modine T, Delhaye C, Rosey G, Gourlay T,
Bauters C, Lablanche JM.
High incidence of recurrent in stent thrombosis after successful treatment
of a first in stent thrombosis.
Catheter Cardiovasc Interv, 2008, 72:470-478.
de Groote P, Gressin V, Hachulla E, Carpentier P, Guillevin L, Kahan A,
Cabane J, Francès C, Lamblin N, Diot E, Patat F, Sibilia J, Petit H,
Cracowski JL, Clerson P, Humbert M ; ItinerAIR-Scleroderma Investigators.
Evaluation of cardiac abnormalities by Doppler echocardiography in a
large nationwide multicentric cohort of patients with systemic sclerosis.
Ann Rheum Dis, 2008, 67:31-36.
Le Tourneau T, Susen S, Caron C, Millaire A, Maréchaux S, Polge AS,
Vincentelli A, Mouquet F, Ennezat PV, Lamblin N, de Groote P,
Van Belle E, Deklunder G, Goudemand J, Bauters C, Jude B.
Functional impairment of von Willebrand factor in hypertrophic
cardiomyopathy: relation to rest and exercise obstruction.
Circulation, 2008, 118:1550-1557.
Dupont A, Chwastyniak M, Beseme O, Guihot AL, Drobecq H,
Amouyel P, Pinet F.
human macrophages.
J Proteome Res, 2008, 7:3572-3582.
Lo T, Sourgounis A, Nolan J, Hamon M.
Starting a transradial programme : How to go about it ?
Ind Heart J, 2008, 60 (Suppl A):A3-A8.
Ennezat PV, Maréchaux S, Huerre C, Deklunder G, Asseman P, Jude B,
Van Belle E, Mouquet F, Bauters C, Lamblin N, Lejemtel TH, de Groote P.
Exercise does not enhance the prognostic value of Doppler
echocardiography in patients with left ventricular systolic dysfunction
and functional mitral regurgitation at rest.
Am Heart J, 2008, 155:752-757.
Maillard-Lefebvre H, Launay D, Mouquet F, Gaxotte V, Hachulla E,
de Groote P, Lambert M, Queyrel V, Morell-Dubois S, Bérégi JP,
Bauters C, Hatron PY.
Polyarteritis nodosa-related coronary aneurysms.
J Rheumatol, 2008, 35:933-934.
Ennezat PV, Darchis J, Lamblin N, Tricot O, Elkohen M, Aumégeat V,
Equine O, Dujardin X, Saadouni H, Le Tourneau T, de Groote P,
Bauters C ; REVE Investigators.
Left ventricular remodeling is associated with the severity of mitral
regurgitation after inaugural anterior myocardial infarction--optimal
timing for echocardiographic imaging.
Am Heart J, 2008, 155:959-965.
Maréchaux S, Pinçon C, Le Tourneau T, de Groote P, Huerre C, Asseman P,
Van Belle E, Nevière R, Bauters C, Deklunder G, Lejemtel TH, Ennezat PV.
Cardiac correlates of exercise induced pulmonary hypertension in patients
with chronic heart failure due to left ventricular systolic dysfunction.
Echocardiography, 2008, 25:386-393.
Mouquet F, Lions C, de Groote P, Bouabdallaoui N, Willoteaux S,
Dagorn J, Deruelle P, Lamblin N, Bauters C, Beregi JP.
Characterisation of peripartum cardiomyopathy by cardiac magnetic
resonance imaging.
Eur Radiol, 2008, 18:2765-2769.
Ennezat PV, Lamblin N, Mouquet F, Tricot O, Quandalle P, Aumégeat V,
Equine O, Nugue O, Segrestin B, de Groote P, Bauters C ; REVE
Investigators.
The effect of ageing on cardiac remodelling and hospitalization for heart
failure after an inaugural anterior myocardial infarction.
Eur Heart J, 2008, 29:1992-1999.
Pinet F, Beseme O, Cieniewski-Bernard C, Drobecq H, Jourdain S,
Lamblin N, Amouyel P, Bauters C.
Proteomics, 2008, 8:1798-1808.
Ennezat PV, Lefetz Y, Maréchaux S, Six-Carpentier M, Deklunder G,
Montaigne D, Bauchart JJ, Mounier-Véhier C, Jude B, Nevière R,
Bauters C, Asseman P, de Groote P, Lejemtel TH.
Left ventricular abnormal response during dynamic exercise in patients
with heart failure and preserved left ventricular ejection fraction at rest.
J Card Fail, 2008, 14:475-480.
Richardson-Lobbedez M, Maréchaux S, Bauters C, Darchis J,
Auffray JL, Bauchart JJ, Aubert JM, Lejemtel TH, Lesenne M,
Van Belle E, Goldstein P, Asseman P, Ennezat PV.
Prognostic importance of tissue Doppler-derived diastolic function in patients
presenting with acute coronary syndrome : a bedside echocardiographic study.
Eur J Echocardiogr, 2008, 9:594-598.
Hamon M, Agostini D, Le Page O, Riddell JW, Hamon M.
Prognostic impact of right ventricular involvement in patients with acute
myocardial infarction. meta-analysis.
Critical Care Med, 2008, 36:2023-2033.
Ritz MF, Ratajczak P, Curin Y, Cam E, Mendelowitsch A, Pinet F,
Andriantsitohaina R.
Chronic treatment with red wine polyphenol compounds mediates
neuroprotection in a rat model of ischemic cerebral stroke.
J Nutr, 2008, 138:519-525.
Hamon M, Sourgounis A.
Radiation exposure and vascular access site.
Eur Heart J, 2008, 29:954.
126
Contents
Hamon M, Rasmussen LH, Manoukian SV, Cequier A, Lincoff AM,
Rupprecht HJ, Gersh BJ, Mann T, Bertrand ME, Mehran R, Stone GW.
Choice of arterial access site and outcomes in patients with acute coronary
syndromes managed with an early invasive strategy : The Acuity trial.
Eurointervention, 2009, 5:115-120.
Valgimigli M, Campo G, de Cesare N, Vranckx P, Hamon M,
Angiolillo D, Sabat M, Ferrari F, Tumscitz C, Repetto A, Meliga E,
Kubbajeh M, Benassi A, Parrinello G, Percoco G, Ferrari R.
Tailoring Treatment with Tirofiban in patients showing Resistance to
aspirin and/or Resistance to clopidogrel (3T/2R) study. Rationale for the
Study and Protocol Design.
Cardiovasc Drugs Ther, 2008, 4:313-320.
Hamon M, Coutance G.
Transradial intervention for minimizing bleeding complications in
percutaneous coronary intervention.
Am J Cardiol. 2009; 104(5 Suppl):55C-59C. Review.
2009
Acosta-Martin AE, Chwastyniak M, Beseme O, Drobecq H,
Amouyel P, Pinet F.
Impact of incomplete DNase I treatment on human macrophage analysis.
Proteomics Clin Appl, 2009, 3:1-11.
Jolly SS, Amlani S, Hamon M, Yusuf S, Mehta SR.
Radial versus Femoral Access for Coronary Angiography or Intervention
and the Impact on Major Bleeding : A Systematic Review and Metaanalysis of Randomized Trials.
Am Heart J, 2009, 157:132-140.
Allouch M, Zhong YZ, Riddell JW, Sabatier R, Hamon M.
Transradial coronary rotational atherectomy using 5-French guiding
catheters.
Chin Med J. (Engl), 2009, 122:1356-1358.
Konoshita T, Kato N, Fuchs S, Mizuno S, Aoyama C, Motomura M,
Makino Y, Wakahara S, Inoki I, Miyamori I, Pinet F.
Genetic variant of the renin-angiotensin system and diabetes influences
blood pressure response to angiotensin receptor blocker.
Diabetes care, 2009, 32:1485-1490.
Clergeau M-R, Hamon M, Morello R, Saloux E, Viader F, Hamon M.
Silent cerebral infarcts in patients with pulmonary embolism and a patent
foramen ovale : a prospective diffusion-weighted magnetic resonnance
imaging study.
Stroke, 2009, 40:3758-3762.
Lemesle G, Delhaye C, Sudre A, Broucqsault D, Rosey G, Bauters C,
Lablanche JM.
Impact of high loading and maintenance dose of clopidogrel within the
first 15 days after percutaneous coronary intervention on patient outcome.
Am Heart J, 2009, 157:375-382.
Delhaye C, Sudre A, Lemesle G, Maréchaux S, Broucqsault D,
Hennache B, Bauters C, Lablanche JM.
Preprocedural high-sensitivity C-reactive protein predicts death or
myocardial infarction but not target vessel revascularization or stent
thrombosis after percutaneous coronary intervention.
Le Tourneau T, Betto M, Richardson M, Juthier F, Ennezat PV, Polge
AS, Bauters C, Vincentelli A, Deklunder G.
Prospective assessment of multiple cardiac paillary fibroelastomas. An
echocardiographic and surgical study.
Int J Cardiol, 2009, Dec 22. [Epub ahead of print]
Am J Hypertens, 2009, 22:993-1000.
Mouquet F, Lamblin N, de Groote P.
Indication for bromocriptine in peripartum cardiomyopathy.
Eur J Heart Fail, 2009, 11:223.
Sourgounis A, Lipiecki J, Lo TS, Hamon M.
Coronary stents and chronic anticoagulation.
Circulation, 2009, 119:1682-1688.
Hachulla AL, Launay D, Gaxotte V, de Groote P, Lamblin N, Devos P,
Hatron PY, Beregi JP, Hachulla E.
Cardiac magnetic resonance imaging in systemic sclerosis : a cross-sectional
observational study of 52 patients.
Ann Rheum Dis, 2009, 68:1878-1884.
Valgimigli M, Campo G, de Cesare N, Meliga E, Vranckx P, Furgieri A,
Angiolillo DJ, Sabatè M, Hamon M, Repetto A, Colangelo S,
Brugaletta S, Parrinello G, Percoco G, Ferrari R ; Tailoring Treatment
With Tirofiban in Patients Showing Resistance to Aspirin and/or
Resistance to Clopidogrel (3T/2R) Investigators.
Intensifying platelet inhibition with tirofiban in poor responders to
aspirin, clopidogrel, or both agents undergoing elective coronary
intervention: results from the double-blind, prospective, randomized
Tailoring Treatment with Tirofiban in Patients Showing Resistance to
Aspirin and/or Resistance to Clopidogrel study.
Circulation, 2009, 119:3215-3222.
Hachulla E, de Groote P, Gressin V, Sibilia J, Diot E, Carpentier P,
Mouthon L, Hatron PY, Jego P, Allanore Y, Tiev KP, Agard C,
Cosnes A, Cirstea D, Constans J, Farge D, Viallard JF, Harle JR,
Patat F, Imbert B, Kahan A, Cabane J, Clerson P, Guillevin L,
Humbert M ; Itinér AIR-Sclérodermie Study Group.
The three-year incidence of pulmonary arterial hypertension associated with
systemic sclerosis in a multicenter nationwide longitudinal study in France.
Arthritis Rheum, 2009, 60:1831-1839.
Hamon M, Mc Fadden E.
Trans-radial approach for cardiovascular interventions (2nd edition).
ESM Editions 2009.
Hachulla E, Bervar JF, Launay D, Lamblin N, Perez T, Mouthon L,
de Groote P, Tillie-Leblond I, Humbert M.
[Dyspnea upon exertion in systemic scleroderma : from symptom to
etiological diagnosis] [Article in French]
Presse Med, 2009, 38:911-926.
2010
Bauters C, Ennezat PV, Lamblin N, de Groote P.
Left ventricular remodeling and heart failure after myocardial infarction
in elderly patients.
Am J Cardiol, 2010, 105:903-904.
Hachulla E, Carpentier P, Gressin V, Diot E, Allanore Y, Sibilia J,
Launay D, Mouthon L, Jego P, Cabane J, de Groote P, Chabrol A,
Lazareth I, Guillevin L, Clerson P, Humbert M ; ItinérAIRSclérodermie Study Investigators.
Risk factors for death and the 3-year survival of patients with systemic
sclerosis: the French ItinérAIR-Sclérodermie study.
Beseme O, Fertin M, Drobecq H, Amouyel P, Pinet F.
Combinatorial peptide ligand library plasma treatment : advantages for
accessing low-abundance proteins.
Electrophoresis, 2010, 31:2697-2704.
127
Contents
Dubois E, Richard V, Mulder P, Lamblin N, Drobecq H, Henry JP,
Amouyel P, Thuillez C, Bauters C, Pinet F.
Decreased Serine207-phosphorylated of troponin T as a biomarker for
left ventricular remodeling after myocardial infarction.
Eur Heart J, 2010, Apr 25. [Epub ahead of print].
Patent
Pinet F, Richard V, Bauters C, Mulder P
Post-translation modified cardiac troponin T as a biomarker of a risk for
heart failure.
EPO9305471.6 du 22 mai 2009
PCT/EP2010/056931 du 20 Mai 2010
Inserm / Institut Pasteur de Lille
Dupont A, Elkalioubie A, Juthier F, Tagzit M, Vincentelli A,
Le Tourneau T, Haulon S, Deklunder G, Breyne J, Susen S,
Marechaux S, Pinet F, Jude B.
Frequency of abdominal aortic aneurysm in patients undergoing
coronary artery bypass grafting.
Am. J Cardiol, 2010, 105:1545-1548.
Fertin M, Beseme O, Duban S, Amouyel P, Bauters C, Pinet F.
Deep plasma proteomic analysis of patients with left ventricular
remodeling after a first myocardial infarction.
Proteomics Clin Appl, 2010a, 4:654-673.
Fertin M, Hennache B, Hamon M, Biausque F, Elkohen M, Nugue O,
Ennezat PV, Tricot O, Lamblin N, Pinet F, Bauters C.
Usefulness of serial assessment of B-type natriuretic peptide, troponin I,
and C-reactive protein to predict left ventricular remodeling after acute
myocardial infarction (From the REVE-2 study).
Am J Cardiol, 2010b, in press.
Drobecq H, Lemoine Y, Koussa M, Amouyel P, Pinet F.
transcriptomic, proteomic and protein array study.
J Prot Res, 2010, 9:3720-3725.
Lemesle G, Sudre A, Bouallal R, Delhaye C, Rosey G, Bauters C,
Lablanche JM.
Impact of thrombus aspiration use and direct stenting on final
myocardial blush score in patients presenting with ST-elevation
myocardial infarction.
Le Tourneau T, Richardson M, Juthier F, Modine T, Fayad G, Polge AS,
Ennezat PV, Bauters C, Vincentelli A, Deklunder G.
Echocardiography predictors and prognostic value of pulmonary artery
systolic pressure in chronic organic mitral regurgiation.
Heart, 2010, 96:1311-1317.
Mekinian A, Lions C, Leleu X, Duhamel A, Lamblin N, Coiteux V,
De Groote P, Hatron PY, Facon T, Beregi JP, Hachulla E, Launay D ;
Lille Amyloidosis Study Group.
Prognosis assessment of cardiac involvement in systemic AL amyloidosis
by magnetic resonance imaging.
Am J Med, 2010, 123(9):864-868
PhD
Adelina Elena ACOSTA MARTIN
Directeur de thèse Florence Pinet
« Recherche de biomarqueurs de l'anévrysme de l'aorte abdominale »
14 décembre 2009
Emilie DUBOIS
Directeur de thèse Christophe Bauters
« Etude de la phosphorylation lors du remodelage ventriculaire postinfarctus dans un modèle expérimental d’insuffisance cardiaque »
18 octobre 2010
128
Contents
However, the discovery of new candidate genes in
dementias still is complex and many results derived from
association studies could not be replicated. In order to
improve our probability of identifying relevant candidate
genes, we combined to our genetic epidemiological
approach an analysis of the variations in gene expression by
a transcriptomic and bio-computing approach. This was how
we selected differentially expressed genes in the brain tissue
of patients against that of control brains. Besides, we
examined alternative splicing events likely to modulate the
biological properties of genes of interest in AD. This latter
approach was developed following the observation that
such events had an impact on the biological functions of
presenilins (proteins involved in early inherited forms of AD).
Search for the molecular
determinants of neurodegenerative diseases
via transcriptomic
and candidate gene
approaches
DR2 Inserm
Group Members :
In order to successfully perform the transcriptional analysis
we first selected all chromosomal regions of interest identified
by positional cloning studies on Alzheimer’s disease. This was
how we could identify more than 463 centimorgans of the
genome distributed among 9 different chromosomes likely to
contain several candidate genes. A bio-computing analysis
made it possible to identify 2,741 genes in these regions. A
DNA chip was designed from these genes and a differential
transcriptomic analysis was made by comparing brain tissues
of AD patients with those from healthy controls. At the same
time, transcriptomic analyses were carried out on the same
tissues using Affymetrix pangenomic chips. All these findings
were thoroughly analysed with bio-computing tools in the
unit, to identify approximately 106 possible differentially
expressed candidate genes.
Richard Florence, MCU Lille 2
Ayral Anne-Marie, Engineer IPL
Grenier-Boley Benjamin, CE IPL
Demiautte Florie, Technician contrat ANR
Hansmannel Franck, Postdoctoral fellow contrat LECMA
Chouraki Vincent, PhD Student Lille 2
Laumet Geoffroy, PhD Student Lille 2
Mounier Anaïs, PhD Student Lille 2
Letronne Florent, Master 2 Student Lille 2
Key words :
Neurodegenerative disease (mild cognitive impairment,
dementia, Alzheimer s disease). Differential transcriptomics.
Differential proteomics. Bioinformatics. Biomarkers. Molecular
epidemiology. Aging. High throughout genomics. Translational
research. Prevention.
In parallel, we have also implemented a bio-computing
analysis procedure to characterise alternative splicing events
not listed in international databases. This work has led us to
make close to 6 million sequence alignments (genomic DNA
and messenger RNA). From 11,231 genes expressed in the
brain tissue showing a strong biological plausibility, we have
selected 3,572 genes likely to contain an extra, non-listed
exon. We eventually restricted the number of genes of interest
to 350 as they were located in all 463 centimorgans of
chromosomal regions of interest identified by positional
cloning studies on Alzheimer’s disease.
The objective of this theme is to identify new environmental
and genetic determinants leading to the alteration of
cognitive functions and to the conversion to dementia. This
project is focused on two aspects:
- Study of the relations between the cardiovascular risk and
dementias,
- the search for candidate genes via transcriptomic and biocomputing approaches (high throughput technologies).
Our research program in cardiovascular and neurodegenerative
diseases have enabled us to gain better insight into these two
conditions. This was how we endeavoured to assess the
relations between the intake of cholesterol-lowering agents
and the risk of developing Alzheimer’s disease. We found that a
decrease in the risk of dementia in subjects treated with lipidreducing agents (Dufouil et al, 2005) and a slower decline of
their MMSE compared to non-treated subjects (Masse et al,
2005). We have also studied several factors of genetic
susceptibility of the cardiovascular risk which may play a role in
the nervous system. This was how we excluded ACE (Bruandet
et al, in the press), APOAI (Helbecque et al, 2008) and VEGF
(Chapuis et al, 2006) genes as genetic determinants of
Alzheimer’s disease. However, we have noted that a
polymorphism of gene MMP-3 was linked to an increase in the
risk of dementia, though only in individuals who do not carry
allele 4 of the APOE gene – a major genetic determinant of
Alzheimer’s disease - (Helbecque et al, 2007). We have also
confirmed that the gene coding for paraoxonase 1 was
potentially a relevant candidate for Alzheimer’s disease
(Chapuis et al, in the press).
After these genes were selected, genetic polymorphisms
contained in these genes were systematically searched for, and
genetic epidemiologic studies were made on our case-controls
studies. Our first approach (using transcriptomic analysis)
enabled us to identify the Ornithine TransCarba-mylase
(Bensemain et al, Mol Psy, 2007), and using the second approach
(bio-computing), we identified another candidate gene, the
S100b (Lambert et al, Mol Psy 2007). In order to accelerate the
identification of new targets, we have developed a new highthroughput customised chip (Affymetrix). This was how we
could analyse 1,156 Tag-SNP’s located in 82 genes, using two
randomly selected sub-samples from 2 independent studies a French and an American one. This work enabled us to identify
a new candidate gene - NF-HEV/IL-33 located in 9p24.1.
Finally, our competences in the field of genetic epidemiology for
Alzheimer’s disease, and the availability of cases-controls studies
have enabled us to take part in the identification of new
potential determinants in cooperation with other teams. So, in
cooperation with two American teams, we were involved in the
identification of gene CALMH1, a trans-membrane glycoprotein
129
Contents
Guyant-Maréchal L, Rovelet-Lecrux A, Goumidi L, Cousin E,
Hannequin D, Raux G, Penet C, Ricard S, Macé S, Amouyel P,
Deleuze JF, Frebourg T, Brice A, Lambert JC, Campion D.
Variations in the APP promoter region and risk to Alzheimer’s disease.
Neurology, 2007, 68:632-633.
controlling concentration levels in calcium ions inside cells,
whose genetic mutation modifies both the concentration in
calcium and production of ab peptides (Dreses-Werringloer et
al, Cell, 2008).
Helbecque N, Cottel D, Hermant X, Amouyel P.
Impact of the matrix metalloproteinase MMP-3 on dementia.
Neurobiol Aging, 2007, 28:1215-1220.
Lambert JC, Ferreira S, Gussekloo J, Christiansen L, Brysbaert B,
Slagboom PE, Cottel D, Petit T, Hauw JJ, Dekosky ST, Richard F, Berr C,
Lendon CL, Kamboh IM, Mann D, Christensen K, Westendorp R, Amouyel P.
Evidence for the association of the S100beta gene with low cognitive
performance and dementia in the elderly.
Mol Psychiatry, 2007, 12:870-880.
Genetic heterogeneity of Alzheimer's disease : Complexity and advances.
Psychoneuroendocrinology, 2007, 32 (Suppl 1):S62-S70.
Leveziel N, Souied EH, Richard F, Barbu V, Zourdani A, Morineau G,
Zerbib J, Coscas G, Soubrane G, Benlian P.
PLEKHA1-LOC387715-HTRA1 polymorphisms and exudative age-related
macular degeneration in the French population.
Mol Vis, 2007, 13:2153-2159.
Considering the countless false-positives, the editors of
journals having a high impact factor significantly raised their
requirements for association studies, and before a manuscript
is accepted, we are often asked to add functionality studies or
experimental in vivo or in vitro analyses. In order to fulfil these
requirements, we work in cooperation with several teams
having expertise in the fields of cell biology such as Inserm
unit 837, in particular. We are also exploring the impact of
these factors on mild to moderate alteration of cognitive
functions in individuals without dementia. These studies
enable us to identify individuals who will eventually suffer
from a conversion to dementia. The impact of the genes so
identified is also assessed in other types of neurodegenerative diseases: fronto-temporal dementia, sporadic
Parkinson’s disease so as to determine if common
neurodegenerative processes may be identified. Finally, the
impact of candidate genes so identified is studied in
epidemiological studies defined in team 1 and team 2 on the
vascular risk (Dumont et al, Am J Hyperten, 2009).
Meirhaeghe A, Boreham CA, Murray LJ, Richard F, Davey Smith G,
Young IS, Amouyel P.
A possible role for the PPARG Pro12Ala polymorphism in preterm birth.
Diabetes, 2007, 56:494-498.
Vasseur F, Guerardel A, Barat-Houari M, Cottel D, Amouyel P,
Froguel P, Helbecque N.
Impact of a CART promoter genetic variation on plasma lipid profile in a
general population.
Mol Genet Metab, 2007, 90:199-204.
Zawadzki C, Susen S, Richard F, Haulon S, Corseaux D, Jeanpierre E,
Vincentelli A, Lucas C, Torpier G, Martin A, Van Belle E, Staels B, Jude B.
Dyslipidemia shifts the tissue factor/tissue factor pathway inhibitor
balance toward increased thrombogenicity in atherosclerotic plaques.
Evidence for a corrective effect of statins.
Atherosclerosis, 2007, 195:e117-e125.
To conclude, in the past four years, we have attained the
objectives which we had set to ourselves at the time of the
creation of the unit, particularly in terms of completion of
epidemiological studies, development of new proteomic,
transcriptomic and bio-computing screening technological
approaches and concerning the identification of determinants
for cardiovascular and neurodegenerative diseases.
Pasquier F, Richard F and Legrain S.
Maladie d’Alzheimer.
In : Traité de Santé Publique 2e édition. Eds : Médecine-Science,
Flammarion, Paris, 2007.
2008
Bruandet A, Richard F, Bombois S, Maurage CA, Masse I,
Amouyel P, Pasquier F.
Cognitive decline and survival in Alzheimer’s disease according to
education level.
Publications
2007
Bruandet A, Richard F, Tzourio C, Berr C, Dartigues JF,
Alpérovitch A, Amouyel P, Helbecque N.
Haplotypes across ACE and the risk of Alzheimer's disease : The Three-City
Study.
J Alzheimers Dis 2008, 13:333-339.
Boddaert J, Kinugawa K, Lambert JC, Boukhtouche F, Zoll J, Merval
R, Blanc-Brude O, Mann D, Berr C, Vilard J, Garabedian B,
Silvestre JS, Duyckaerts C, Amouyel P, Mariani J, Tedgui A, Mallat Z.
Evidence of a role for lactadherin in Alzheimer's disease.
Am J Pathol, 2007, 170:921-929.
Campagne F, Lambert JC, Dreses-Werringloer U, Vingtdeux V,
Lendon C, Campion D, Amouyel P, Lee AT, Gregersen PK, Davies P,
Marambaud P.
CALHM1 association with Alzheimer's disease risk response.
Cell, 2008, 135:994-996. Response.
Gutierrez-Aguilar R, Benmezroua Y, Balkau B, Marre M, Helbecque
N, Charpentier G, Polychronakos C, Sladek R, Froguel P, Neve B.
Minor contribution of Smad7 and KLF10 variants to genetic susceptibility
of type 2 diabetes.
Diabetes Metab, 2007, 33:372-378.
130
Contents
Hauw JJ, DeKosky ST, Lemoine Y, Iwatsubo T, Wavrant-De Vrièze F,
Dartigues JF, Tzourio C, Buee L, Pasquier F, Berr C, Mann D,
Lendon C, Alpérovitch A, Kamboh MI, Amouyel P, Lambert JC.
Transcriptomic and genetic studies Identify IL-33 as a candidate gene for
Mol Psychiatry, 2009, 14:1004-1016.
Chapuis J, Hannequin D, Pasquier F, Bentham P, Brice A, Leber I,
Frebourg T, Deleuze JF, Cousin E, Thaker U, Amouyel P, Mann D,
Lendon C, Campion D, Lambert JC.
Association study of the GAB2 gene with the risk of developing
Neurobiol Dis, 2008, 30:103-106.
Lambert JC, Campagne F, Marambaud P.
ALHM1, a novel gene to blame in Alzheimer disease. [Article in French].
Med Sci (Paris), 2008, 24:923-924.
Deschaintre Y, Richard F, Leys D, Pasquier F.
Treatment of vascular risk factors is associated with slower decline in
Neurology, 2009, 73:674-680.
Chapuis J, Moisan F, Mellick G, Elbaz A, Pasquier F, Hannequin D,
Lendon C, Campion D, Amouyel P, Lambert JC.
Association study of the NEDD9 gene with the risk of developing
Alzheimer's and Parkinson's disease.
Hum Mol Genet, 2008, 17:2863-2867.
Am J Hypertens, 2009, 22:993-1000.
Dreses-Werringloer U, Lambert JC, Vingtdeux V, Zhao H, Vais H,
Siebert A, Jain A, Koppel J, Rovelet-Lecrux A, Hannequin D,
Pasquier F, Galimberti D, Scarpini E, Mann D, Lendon C, Campion D,
Amouyel P, Davies P, Foskett JK, Campagne F, Marambaud P.
A polymorphism in CALHM1 influences Ca2+ homeostasis, Abeta levels,
and Alzheimer's disease risk.
Cell 2008, 133:1149-1161.
Goumidi L, Flamant F, Lendon C, Galimberti D, Pasquier F,
Scarpini E, Hannequin D, Campion D, Amouyel P, Lambert JC,
Meirhaeghe A.
An apolipoprotein A-I gene promoter polymorphism associated with
cognitive decline, but not with Alzheimer's disease.
Hansmannel F, Lendon C, Pasquier F, Dumont J, Hannequin D,
Chapuis J, Laumet G, Ayral AM, Galimberti D, Scarpini E,
Is the ornithine transcarbamylase gene a genetic determinant of
Alzheimer's disease ?
Neurosci Lett, 2009, 449:76-80.
Le Ber I, Camuzat A, Hannequin D, Pasquier F, Guedj E, RoveletLecrux A, Hahn-Barma V, van der Zee J, Clot F, Bakchine S, Puel M,
Ghanim M, Lacomblez L, Mikol J, Deramecourt V, Lejeune P,
de la Sayette V, Belliard S, Vercelletto M, Meyrignac C,
Broeckhoven CV, Lambert JC, Verpillat P, Campion D, Habert MO,
Dubois B, Brice A ; the French research network on FTD/FTD-MND.
Phenotype variability in progranulin mutation carriers: a clinical,
neuropsychological, imaging and genetic study.
Brain, 2008, 131:732-746.
An age effect on the association of common variants of ACE with
Neurosci Lett, 2009, 461:181-184.
Helbecque N, Cottel D, Amouyel P.
Low-density lipoprotein receptor-related protein 8 gene polymorphisms
and dementia.
Leveziel N, Zerbib J, Richard F, Querques G, Morineau G, FremeauxBacchi V, Coscas G, Soubrane G, Benlian P, Souied EH.
Genotype-phenotype correlations for exudative Age-related Macular
Degeneration associated with homozygous HTRA1 and CFH genotypes.
Association of plasma amyloid {beta} with risk of dementia : The
prospective Three-City Study.
Neurology, 2009, 73:847-853.
Rovelet-Lecrux A, Deramecourt V, Legallic S, Maurage CA, Le Ber I,
Brice A, Lambert JC, Frébourg T, Hannequin D, pasquier F, Campion D.
Deletion of the progranulin gene in patients with frontotemporal lobar
degeneration of Parkinson disease.
Neurobol Dis, 2008, 31:41-45.
Lambert JC, Heath S, Even G, Campion D, Sleegers K, Hiltunen M,
Combarros O, Zelenika D, Bullido MJ, Tavernier B, Letenneur L,
Bettens K, Berr C, Pasquier F, Fiévet N, Barberger-Gateau P,
Engelborghs S, De Deyn P, Mateo I, Franck A, Helisalmi S,
Porcellini E, Hanon O ; the European Alzheimer's Disease Initiative
Investigators, de Pancorbo MM, Lendon C, Dufouil C, Jaillard C,
Leveillard T, Alvarez V, Bosco P, Mancuso M, Panza F, Nacmias B,
Bossù P, Piccardi P, Annoni G, Seripa D, Galimberti D, Hannequin D,
Licastro F, Soininen H, Ritchie K, Blanché H, Dartigues JF, Tzourio C,
Gut I, Van Broeckhoven C, Alpérovitch A, Lathrop M, Amouyel P.
Nat Genet, 2009, 41:1094-1099.
2009
Bensemain F, Hot D, Ferreira S, Dumont J, Bombois J, Maurage CA,
Lemoine Y, Hauw JJ, Schraen S, Lemoine Y, Buée L, Berr C, Mann D,
Pasquier F, Amouyel P, Lambert JC.
Mol Psychiatry, 2009, 14:106-116.
Bruandet A, Richard F, Bombois S, Maurage CA, Deramecourt V,
Lebert F, Amouyel P, Pasquier F.
Alzheimer's disease with cerebrovascular disease and vascular dementia :
clinical features and course compared with Alzheimer's disease.
J Neurol Neurosurg Psychiatry, 2009, 80:133-139.
Sparks DL, Hunsaker JC 3rd, Amouyel P, Malafosse A, Bellivier F,
Leboyer M, Courtet P, Helbecque N.
Angiotensin I-converting enzyme I/D polymorphism and suicidal behaviors.
Chapuis J, Boscher M, Bensemain F, Cottel D, Amouyel P, Lambert JC.
Association study of the paraoxonase 1 gene with the risk of developing
131
Contents
Laumet G, Chouraki V, Grenier Boley B, Legry V, Heath S, Zelenika D,
Epelbaum J, Berr C, Dartigues J-F, Tzourio C, Campion D, Lathrop M,
Systematic analysis of candidate genes for Alzheimer’s disease in a
French, genome-wide association study.
J Alzheimers Dis, 2010, 20:1181-1188.
Zerbib J, Seddon JM, Richard F, Reynolds R, Leveziel N, Benlian P, Borel P,
Feingold J, Munnich A, Soubrane G, Kaplan J, Rozet JM, Souied EH.
rs5888 variant of SCARB1 gene is a possible susceptibility factor for agerelated macular degeneration.
PLoS One, 2009, 4:e7341.
2010
Le Fur I, Laumet G, Richard F, Fievet N, Berr C, Rouaud O, Delcourt C,
Association study of the CFH Y402H polymorphism with Alzheimer's disease.
Goumidi L, Flamant F, Lendon C, Galimberti D, Pasquier F, Scarpini E,
Hannequin D, Campion D, Amouyel P, Lambert JC, Meirhaeghe A.
Leveziel N, Puche N, Richard F, Somner JE, Zerbib J, Bastuji-Garin S,
Cohen S, Korobelnik JF, Sahel JA, Soubrane G, Benlian P, Souied EH.
Genotypic influences on severity of exudative Age-related Macular
Degeneration.
Hansmannel F, Sillaire A, Kamboh MI, Lendon C, Pasquier F,
Hannequin D, Laumet G, Mounier A, Ayral A-M, Dekosky ST,
Hauw J-J, Berr C, Mann D, Amouyel P, Campion D, Lambert JC.
Is the urea cycle involved in Alzheimer's disease ?
J Alzheimers Dis, 2010, 21:1013-1021.
Seshadri S, Fitzpatrick AL, Ikram A, DeStefano AL, Gudnason V,
Boada M, Bis JC, Smith AV, Carassquillo MM, Lambert JC, Harold D,
Schrijvers EMC, Ramirez-Lorca R, Debette S, Longstreth WT,
Janssens CJW,Pankratz, Dartigues J-F, Hollinworth P, Aspelund T,
Hernadez I, Beiser A, Kuller LH, Koudstaal PJ, Dickson DW, Tzourio
C, Abraham R, Antunez C, Du, Y, Rotter JI, Aulchenko YS, Harris TB,
Petersen RC, Berr C, Owen MJ, Lopez-Arrieta J, Varadarajan BN,
Becker JT, Rivadeneira F, Nalls MA, Graff-Radford NR, Campion D,
Auerbach S, Rice K, Hofman A, Jonsson PV, Schmidt H, Lathrop M,
Mosley TH, Au R, Psaty BM, Uitterlinden AG, Farrer LA, Lumley T, Ruiz A,
Williams J, Amouyel P, Younkin SG, Wolf PA, Launer LJ, Lopez O, Van Duijn C,
Breteler M on behalf of the Charge, GERAD1 and EADI1 consortia.
Genome-wide association studies of alzheimer’s disease.
JAMA, 2010, 303:1864-65.
Hong MG, Alexeyenko A, Lambert JC, Amouyel P, Prince JA.
Genome-wide pathway analysis implicates intracellular transmembrane
protein transport in Alzheimer disease.
J Hum Genet, 2010, in press.
Krüger R, Sharma M, Riess O, Gasser T, Van Broeckhoven C, Theuns J,
Aasly J, Annesi G, Bentivoglio AR, Brice A, Djarmati A, Elbaz A, Farrer M,
Ferrarese C, Gibson JM, Hadjigeorgiou GM, Hattori N, Ioannidis JP,
Jasinska-Myga B, Klein C, Lambert JC, Lesage S, Lin JJ, Lynch T,
Mellick GD, de Nigris F, Opala G, Prigione A, Quattrone A, Ross OA,
Satake W, Silburn PA, Tan EK, Toda T, Tomiyama H, Wirdefeldt K,
Wszolek Z, Xiromerisiou G, Maraganore DM ; for the Genetic
Epidemiology of Parkinson's disease consortium.
A large-scale genetic association study to evaluate the contribution of
Omi/HtrA2 (PARK13) to Parkinson's disease.
Sleegers K, Lambert JC, Bertram L, Cruts M, Amouyel P,
Van Broeckhoven C.
The pursuit of susceptibility genes for Alzheimer's disease : progress and
prospects.
Trends Genet, 2010, 26:84-93.
Deciphering genetic susceptibility to frontotemporal lobar dementia.
Nat Genet, 2010, 42:189-190.
Berr C, Dartigues J-F, Tzourio C, Campion D, Lathrop M, Amouyel P.
Implication of the immune system in Alzheimer’s disease : evidence from
genome-wide pathway analysis.
J Alzheimers Dis, 2010, 20:1107-1118.
Zerbib J, Richard F, Puche N, Leveziel N, Cohen SY, Korobelnik JF,
Sahel J, Munnich A, Kaplan J, Rozet JM, Souied EH.
R102G polymorphism of the C3 gene associated with exudative agerelated macular degeneration in a French population.
Mol Vis, 2010, 16:1324-1330.
Lambert JC, Sleegers K, González-Pérez A, Ingelsson M, Beecham GW,
Hiltunen M, Combarros O, Bullido MJ, Brouwers N, Bettens K, Berr C,
Pasquier F, Richard F, Dekosky ST, Hannequin D, Haines JL, Tognoni G,
Fiévet N, Dartigues JF, Tzourio C, Engelborghs S, Arosio B, Coto E,
De Deyn P, Del Zompo M, Mateo I, Boada M, Antunez C, LopezArrieta J, Epelbaum J, Schjeide BM, Frank-Garcia A, Giedraitis V,
Helisalmi S, Porcellini E, Pilotto A, Forti P, Ferri R, Delepine M,
Zelenika D, Lathrop M, Scarpini E, Siciliano G, Solfrizzi V, Sorbi S,
Spalletta G, Ravaglia G, Valdivieso F, Vepsäläinen S, Alvarez V, Bosco P,
Mancuso M, Panza F, Nacmias B, Bossù P, Hanon O, Piccardi P,
Annoni G, Mann D, Marambaud P, Seripa D, Galimberti D, Tanzi RE,
Bertram L, Lendon C, Lannfelt L, Licastro F, Campion D, PericakVance MA, Soininen H, Van Broeckhoven C, Alpérovitch A, Ruiz A,
Kamboh MI, Amouyel P.
The CALHM1 P86L polymorphism is a genetic modifier of age at onset in
Alzheimer's Disease : a meta-analysis study.
J Alzheimers Dis, 2010, in press.
PhD
Julien CHAPUIS
Directeur de thèse Jean-Charles Lambert
« Identification de déterminants génétiques impliqués dans la composante
vasculaire de la MA, par analyses transcriptomiques, génétiques et
moléculaires
25 septembre 2008
Patent
Method for determining the risk of occurrence of Alzheimer’s disease
IPL - Inserm - CEA - Université Lille 2
25 septembre 2008
Laumet G, Petitprez V, Sillaire A, Ayral AM, Hansmannel F, Chapuis
J, Hannequin D, Pasquier F, Scarpini E, Galimberti D, Lendon C,
A study of the association between the ADAM12 and SH3PXD2A
(SH3MD1) genes and Alzheimer's disease.
Neurosci Lett, 2010, 468:1-2.
132
Contents
Genomics
and Metabolic Diseases
CNRS UMR 8199
Philippe FROGUEL
Contact :
00 33 3 20 87 79 54
[email protected]
Group Members :
Froguel Philippe, DR CNRS
Stehelin Dominique, DR CNRS
Wolowzcuk Isabelle, DR2 CNRS
Bouatia-Naji Nabila, CR Inserm
Cauchi Stéphane, CR CNRS
Meyre David, CR Inserm
Neve Bernadette, CR Inserm
Poulain-Godefroy Odile, CR IPL
Vaxillaire Martine, CR IPL
Chèvre Jean-Claude, IR CNRS
Delplanque Jérôme, IR CNRS
Gaget Stefan, IR CNRS
Gallina Sophie, IR CNRS
Lecoeur Cécile, IR CNRS
Sand Olivier, IR CNRS
Yengo Loïc, IR CNRS (CDD)
Couturier Cyril, MCU Lille 2
Allegaert Frédéric, AI Lille 2 (CDD)
Durand-Charton Emmanuelle, AI CNRS
Huyvaert Marlène, AI CNRS (CDD)
Khezami Hager, AI CNRS
Leloire Audrey, AI CNRS
Lobbens Stéphane, AI CNRS
Beury Delphine, IE CNRS (CDD)
Cavalcanti-Proença Christine, IE CNRS (CDD)
Dechaume Aurélie, IE CNRS (CDD)
De Graeve Franck, IE CNRS (CDD)
Eury Elodie, IE CNRS (CDD)
Labrune Yann, IE CNRS (CDD)
Le Guilcher David, IE CNRS
Marchand Marion, IE CNRS (CDD)
Tarront Solène, IE CNRS (CDD)
Vaillant Emmanuel, IE CNRS
Vivequin Sidonie, IE CNRS (CDD)
Delfosse Philippe, Technician IPL
Deweirder Marianne, Technician IPL
Flament Nicolas, Technician (CDD)
Gallina Philippe, Technician IPL
Hocquet Mélanie, Administrative staff IPL
Poulain Sylvie, Technician CNRS
Vatin Vincent, Technician IPL
Le Bacquer Olivier, Postdoctoral fellow
Bacart Johan, PhD Student IPL/région
Bonnefond Amélie, PhD Student CNRS/région
Choquet Hélène, PhD Student
Marquez Marcel, PhD Student CHRU/Région
Morandi Anita, PhD Student co-tutelle
Key Words :
Type 2 Diabetes. MODY. Obesity. Bioinformatics. Genotyping.
Sequencing. Candidate genes
133
Contents
A - Research Report
development 2010
The UMR 8199-Lille2-CNRS-IPL unit “Genomics and molecular
physiology of metabolic diseases” has been created 4 years ago
with three main objectives : 1/ to understand the contribution
of genes predisposing or associated with type 2 diabetes
(T2D) or obesity risk to the physiology of these disorders, 2/
to develop genetic epidemiology approaches, and 3/ to
contribute to elucidate the molecular determinants of traits
related to glucose homeostasis.
In the last 18 months, outstanding progress has been made
in the knowledge of the genetic background of T2D (“a
breakthrough of the year 2007” according to Science). With the
development of GWA approaches using high density SNP
microarrays, we indeed completed and published in February
2007 in Nature the first two-stage GWA study of T2D (5th best
referred paper in the world science in 2007). Subsequently,
our post-GWA analyses showed the additive effect of the
newly discovered variants to the risk of T2D, and their
predictive interest in the general population. We also
proposed that novel T2D GWA analyses should study the
contribution of genome variation to quantitative traits (QT)
associated with glucose homeostasis. This approach led us to
publish in Science in spring 2008 that G6PC2/IGRP modulates
fasting plasma glucose values, and very recently we identified
the role of the melatonin receptor 1B in the maintenance of
glycemia and also in the risk to T2D. In monogenic T2D we
found in 2006 that the sulfonylurea receptor (SUR1/ABCC8)
may cause neonatal diabetes, a discovery that contributed to
one of most striking evidence of the interest of genomic
medicine and pharmacogenetics so far.
T2D and obesity may have common bases. In line with this,
our study of the ENPP1-PC1 encoding for an inhibitor of the
insulin receptor showed evidence for a role of variation in this
gene in both severe early onset obesity and diabetes. We also
showed genetic and functional evidence for a contribution to
the obesity risk of frequent non synonymous mutations in
PCSK1(encoding for the proconvertase 1, a key enzyme for the
maturation of several neuropetides involved in appetite
regulation). We have also revisited the contribution of the the
known obesity gene locus MC4R, which led us to publish
several papers that illustrate the range of physiological effects
of genetic variation in this biologically important gene on
both monogenic and polygenic obesity. In obesity research,
our major discovery was the identification in 2007 of the gene
named “FaT and Obesity associated gene” (FTO), which seems
to be the most potent polygenic obesity gene identified so far
but the function of this gene in human remains elusive.
Another major achievement of the laboratory in 2007 was the
publication of a PNAS paper showing the role of the tetraspan
protein OB-RGRP in the leptin receptor trafficking and the
effect of genetic manipulation in rodents for appetite
regulation.
In conclusion, we have fully achieved our initial objectives. We
have taken advantage of the most recent striking
methodological progress and have been instrumental in
several of the recent discoveries in the genetics of T2D and
obesity.
We are now seeking at establishing an integrative genome
variation map of T2D and Obesity, using state-of-the-art
genomic approaches using our comprehensive genome core
lab for high throughput DNA analysis. Our project includes
several complementary directions that will be followed in
close collaboration with our Imperial College London twin
laboratory and also with other partners (in Lille, Paris and
abroad). We have two closely linked scientific projects, on T2D
and on obesity. We want :
1/ to develop a “deep T2D GWA analysis” combining several
genome-wide approaches to discover novel T2D loci and the
causative variants in already strongly associated loci,
2/ to evaluate in large European population-based studies the
effects attributable to these etiologic genetic variations, as
well as their possible interaction with environment,
3/ to integrate both transcriptomics, SNP/CNV/rare
variants/epigenetic DNA data, with extensive phenotypic
data (dichotomous, and clinical and biological quantitative
traits) from diabetics and controls,
4/ to elucidate novel forms and novel molecular mechanisms
involved in monogenic diabetes (MODYx, neonatal and
early-infancy diabetes), and
5/ to develop synergetic collaborative approaches in molecular
and cellular biology to study functional consequences of the
most significant T2D-associated genes, gene variants, and
genes regulated by T2D-associated transcription factors (e.g.
TCF7L2).
We have recently completed a GWA study of severe obesity.
We now propose :
1/ to apply a combined (meta-analysis) high density genomewide SNP association scan in extreme obesity in a total of
7,847 normal-weight controls and 4,660 extremely obese
subjects of European origin,
2/ to evaluate the contribution of the most promising SNP
associations with extreme obesity in different populations
(24,265 samples),
3/ to conduct deep analyses in the best promising new obesity
loci in order to identify the obesity causative SNPs (e.g. by
fine-mapping the FTO locus),
4/ to evaluate the predictive, cumulative and synergetic effects
of obesity associated SNPs, and SNPs-environment
interactions in large case-control and population-based
prospective cohorts,
134
Contents
Chu WS, Das SK, Wang H, Chan JC, Deloukas P, Froguel P, Baier LJ,
Jia W, Mccarthy MI, Ng MC, Damcott C, Shuldiner AR, Zeggini E,
Elbein SC.
Activating Transcription Factor 6 (ATF6) Sequence Polymorphisms in Type
2 Diabetes and Pre-Diabetic Traits.
Diabetes, 2007, 56:856-862.
5/ to explore the physiological and molecular mechanisms by
which they may contribute to obesity by studying the
correlation between genotypes and mRNA expression in
adipose tissue and muscle of obese patients, and the
obesity-associated intermediary traits in both obesityselected and general populations, and by searching for
mutations in monogenic forms of obesity/leanness in the
new obesity susceptibility genes, and 6/ to explore the
function of proteins related to obesity (i.e. OB-RGRP and
LEPROTL1).
De Smith AJ, Tsalenko A, Sampas N, Scheffer A, Yamada NA,
Tsang P, Ben-Dor A, Yakhini Z, Ellis RJ, Bruhn L, Laderman S,
Froguel P, Blakemore AI.
Array CGH analysis of copy number variation identifies 1284 new genes
variant in healthy white males : implications for association studies of
complex diseases.
Hum Mol Genet, 2007, 16:2783-2794.
Our overall goal in the next years is the sustainable
development of a genomic “environment” in Lille around
metabolic diseases. Together with 2 highly regarded
Inserm/Lille 2 University groups we will establish the European
Genomics Institute for Diabetes research (EGID). We want to
develop “system biology” approaches of diabetes and related
diseases, and to share state-of-the-art platforms for human
and animal structural and functional genomics of metabolic
diseases. This novel institute should attract several additional
teams (through a “hotel à projet” structure) in order to achieve
a critical mass in diabetes research.
Dina C, Meyre D, Gallina S, Durand E, Korner A, Jacobson P,
Carlsson Lm, Kiess W, Vatin V, Lecoeur C, Delplanque J, Vaillant E,
Pattou F, Ruiz J, Weill J, Levy-Marchal C, Horber F, Potoczna N,
Hercberg S, Le Stunff C, Bougneres P, Kovacs P, Marre M, Balkau B,
Cauchi S, Chevre JC, Froguel P.
Variation In FTO Contributes To Childhood Obesity And Severe Adult
Obesity.
Nat Genet, 2007, 39:724-726.
Dina C, Meyre D, Samson C, Tichet J, Marre M, Jouret B, Charles Ma,
Balkau B, Froguel P.
Comment on "A Common Genetic Variant Is Associated with Adult and
Childhood Obesity”.
Science, 2007, 315:187.
Publications
Fairbrother UL, Tanko LB, Walley AJ, Christiansen C, Froguel P,
Blakemore AI.
Leptin Receptor Genotype at Gln223Arg is Associated with Body
Composition, Bone Mineral Density and Vertebral Fracture in
Postmenopausal Danish Women.
J Bone Miner Res, 2007, 22:544-550.
2007
Bell CG, Meyre D, Petretto E, Levy-Marchal C, Hercberg S,
Charles Ma, Boyle C, Weill J, Tauber M, Mein CA, Aitman TJ,
Froguel P, Walley AJ.
No contribution of angiotensin-converting enzyme (ACE) gene variants
to severe obesity : a model for comprehensive case/control and
quantitative cladistic analysis of ACE in human diseases.
Eur J Hum Genet, 2007, 15:320-327.
Fanciulli M, Norsworthy PJ, Petretto E, Dong R, Harper L, Kamesh L,
Heward JM, Gough SC, De Smith A, Blakemore AI, Froguel P,
Owen CJ, Pearce ShHTeixeira L, Guillevin L, Graham DS, Pusey CD,
Cook HT, Vyse TJ, Aitman TJ.
FCGR3B copy number variation is associated with susceptibility to
systemic, but not organ-specific, autoimmunity.
Nat Genet, 2007, 39:721-723.
Bouatia-Naji N, Vatin V, Lecoeur C, Heude B, Proenca C, Veslot J,
Jouret B,Tichet J, Charpentier G, Marre M, Balkau B, Froguel P,
Meyre D.
Secretory granule neuroendocrine protein 1 (SGNE1) genetic variation
and glucose intolerance in severe childhood and adult obesity.
Flechtner I, Vaxillaire M, Cave H, Froguel P, Polak M.
Arch Pediatr, 2007, 14(11):1356-1365.
Ghoussaini M, Vatin V, Lecoeur C, Abkevich V, Younus A, Samson C,
Wachter C, Heude B, Tauber M, Tounian P, Hercberg S, Weill J, LevyMarchal C, Le Stunff C, Bougneres P, Froguel P, Meyre D.
Genetic Study of the Melanin-Concentrating Hormone Receptor 2
(MCHR2) in Childhood and Adulthood Severe Obesity.
J Clin Endocrinol Metab, 2007, 92:4403-4409.
Cauchi S, Vaxillaire M, Choquet H, Durand E, Duval A, Polak M,
Froguel P.
No major contribution of TCF7L2 sequence variants to maturity onset of
diabetes of the young (MODY) or neonatal diabetes mellitus in French
white subjects.
Diabetologia, 2007, 50:214-216.
Ghoussaini M, Vatin V, Lecoeur C, Abkevich V, Younus A, Samson C,
Wachter C, Heude B, Tauber M, Tounian P, Hercberg S, Weill J, LevyMarchal C, Le Stunff C, Bougneres P, Froguel P, Meyre D.
Genetic Study of the Melanin-Concentrating Hormone Receptor 2
(MCHR2) in Childhood and Adulthood Severe Obesity.
Cauchi S, El Achhab Y, Choquet H, Dina C, Krempler F, Weitgasser R,
Nejjari C, Patsch W, Chikri M, Meyre D, Froguel P.
TCF7L2 Is Reproducibly Associated With Type 2 Diabetes In Various Ethnic
Groups: A Global Meta-Analysis.
J Mol Med, 2007, 85:777-782.
Cauchi S, Meyre D, Choquet H, Deghmoun S, Durand E, Gaget S,
Lecoeur C, Froguel P, Levy-Marchal C.
TCF7L2 rs7903146 variant does not associate with smallness for
gestational age in the French population.
Gueorguiev M, Wiltshire S, Garcia EA, Mein C, Lecoeur C, Kristen B,
Allotey R, Hattersley AT, Walker M, O'rahilly S, Froguel P,
Grossman AB, Mccarthy MI, Hitman GA, Korbonits M.
Examining The Candidacy of Ghrelin as a Gene Responsible For Variation
in Adult Stature in a UK Population With Type 2 Diabetes.
135
Contents
Gutierrez-Aguilar R, Benmezroua Y, Balkau B, Marre M, Helbecque N,
Charpentier G, Polychronakos C, Sladek R, Froguel P, Neve B.
Minor contribution of SMAD7 and KLF10 variants to genetic susceptibility
of type 2 diabetes.
Diabetes Metab, 2007, 33:372-378.
Saunders CL, Chiodini BD, Sham P, Lewis CM, Abkevich V,
Adeyemo AA, De Andrade M, Arya R, Berenson GS, Blangero J,
Boehnke M, Borecki IB, Chagnon Yc, Chen W, Comuzzie AG,
Deng HW, Duggirala R, Feitosa MF, Froguel P, Hanson RL,
Hebebrand J, Huezo-Dias P, Kissebah AH, Li W, Luke A, Martin LJ,
Nash M, Ohman M, Palmer LJ, Peltonen L, Perola M, Price RA,
Redline S, Srinivasan SR, Stern MP, Stone S, Stringham H, Turner S,
Wijmenga C, A Collier D.
Meta-Analysis of Genome-wide Linkage Studies in BMI and Obesity.
Obesity (Silver Spring), 2007, 15:2263-2275.
Gutierrez-Aguilar R, Benmezroua Y, Vaillant E, Balkau B, Marre M,
Charpentier G, Sladek R, Froguel P, Neve B.
Analysis of KLF transcription factor family gene variants in type 2
diabetes.
Siddiq A, Gueorguiev M, Samson C, Hercberg S, Heude B, LevyMarchal C,Jouret B, Weill J, Meyre D, Walley A, Froguel P.
Single nucleotide polymorphisms in the neuropeptide Y2 receptor
(NPY2R) gene and association with severe obesity in French white
subjects.
Diabetologia, 2007, 50:574-584.
Horikoshi M, Hara K, Ito C, Shojima N, Nagai R, Ueki K, Froguel P,
Kadowaki T.
Variations in the HHEX gene are associated with increased risk of type 2
diabetes in the Japanese population.
Diabetologia, 2007, 50:2461-2466.
Horikoshi M, Hara K, Ito C, Nagai R, Froguel P, Kadowaki T.
A genetic variation of the transcription factor 7-like 2 gene is associated
with risk of type 2 diabetes in the Japanese population.
Diabetologia, 2007, 50:747-751.
Sladek R, Rocheleau G, Rung J, Dina C, Shen L, Serre D, Boutin P,
Vincent D, Belisle A, Hadjadj S, Balkau B, Heude B, Charpentier G,
Hudson TJ, Montpetit A, Pshezhetsky AV, Prentki M, Posner BI,
Balding DJ, Meyre D, Polychronakos C, Froguel P.
A Genome-Wide Association Study Identifies Novel Risk Loci For Type 2
Diabetes.
Nature, 2007, 445:881-885.
Hummel M, Vasseur F, Mathieu C, Bellanne-Chantelot C, Froguel P,
Standl E, Fuchtenbusch M.
Two Caucasian Families with the Hepatocyte Nuclear Factor-1Alpha
Mutation Tyr218Cys.
Exp Clin Endocrinol Diabetes, 2007, 115:62-64.
Stutzmann F, Vatin V, Cauchi S, Morandi A, Jouret B, Landt O,
Tounian P, Levy-Marchal C, Buzzetti R, Pinelli L, Balkau B, Horber F,
Bougneres P, Froguel P, Meyre D.
Non synonymous polymorphisms in Melanocortin-4 receptor protect
against obesity : the two facets of a Janus obesity gene.
Hum Mol Genet, 2007, 16:1837-1844.
Meyre D, Bouatia-Naji N, Vatin V, Veslot J, Samson C, Tichet J,
Marre M, Balkau B, Froguel P.
ENPP1 K121Q polymorphism and obesity, hyperglycaemia and type 2
diabetes in the prospective DESIR Study.
Diabetologia, 2007, 50:2090-2096.
Valayannopoulos V, Vaxillaire M, Aigrain Y, Jaubert F, BellanneChantelot C, Ribeiro Mj, Brunelle F, Froguel P, Robert JJ, Polak M,
Nihoul-Fekete C, De Lonlay P.
Coexistence in the Same Family of Both Focal and Diffuse Forms of
Hyperinsulinism.
Diabetes Care, 2007, 30:1590-1592.
Owen KR, Groves CJ, Hanson RL, Knowler WC, Shuldiner AR,
Elbein SC, Mitchell BD, Froguel P, NG MC, Chan JC, Jia W,
Deloukas P, Hitman GA, Walker M, Frayling TM, Hattersley AT,
Zeggini E, Mccarthy MI.
Common Variation in the LMNA Gene (Encoding Lamin A/C) and Type 2
Diabetes : Association Analyses in 9,518 Subjects.
Diabetes, 2007, 56:879-883.
Vanbesien-Mailliot CCA, Wolowczuk I, Mairesse J, Viltart O,
Delacre M, Khalife J, Chartier-Harlin MC, Maccari S.
Prenatal stress has pro-inflammatory consequences on the immune
system in adult rats.
Psychoneuroendocrinology, 2007, 32:114-124.
Porzio O, Massa O, Cunsolo V, Colombo C, Malaponti M, Bertuzzi F,
Hansen T, Johansen A, Pedersen O, Meschi F, Terrinoni A, Melino G,
Federici M, Decarlo N, Menicagli M, Campani D, Marchetti P,
Ferdaoussi M, Froguel P, Federici G, Vaxillaire M, Barbetti F.
Missense mutations in the TGM2 gene encoding transglutaminase 2 are
found in patients with early-onset type 2 diabetes.
Mutation in brief no. 982 Hum Mutat, 2007,28:1150.
Vaxillaire M, Dechaume A, Busiah K, Cave H, Pereira S, Scharfmann R,
Perez De Nanclares G, Castano L, Froguel P, Polak M.
New ABCC8 Mutations in Relapsing Neonatal Diabetes and Clinical
Features.
Diabetes, 2007, 56:1737-1741.
Poulain-Godefroy O, Froguel P.
Preadipocyte response and impairment of differentiation in an
inflammatory environment.
Yamauchi T, Nio Y, Maki T, Kobayashi M, Takazawa T, Iwabu M,
Okada-Iwabu M, Kawamoto S, Kubota N, Kubota T, Ito Y, Kamon J,
Tsuchida A, Kumagai K, Kozono H, Hada Y, Ogata H, Tokuyama K,
Tsunoda M, Ide T, Murakami K, Awazawa M, Takamoto I, Froguel P,
Hara K, Tobe K, Nagai R, Ueki K, Kadowaki T.
Targeted Disruption of AdipoR1 and AdipoR2 Causes Abrogation of
Adiponectin Binding and Metabolic Actions.
Nat Med, 2007, 13:332-339.
Salonen JT, Uimari P, Aalto JM, Pirskanen M, Kaikkonen J,
Todorova B, Hypponen J, Korhonen VP, Asikainen J, Devine C,
Tuomainen TP, Luedemann J, Nauck M, Kerner W, Stephens RH,
New JP, Ollier WE, Gibson JM, Payton A, Horan MA, Pendleton N,
Mahoney W, Meyre D, Delplanque J, Froguel P, Luzzatto O, Yakir B,
Darvasi A.
Type 2 diabetes whole-genome association study in four populations: the
DiaGen consortium.
Am J Hum Genet, 2007,81:338-345.
2008
Badii R, Bener A, Zirie M, Al-Rikabi A, Simsek M, Al-Hamaq AO,
Ghoussaini M, Froguel P, Wareham NJ.
Lack of association between the Pro(12)Ala polymorphism of the PPARgamma2 gene and type 2 diabetes mellitus in the Qatari consanguineous
population.
Acta Diabetol, 2008, 45:15-21.
136
Contents
Balkau B, Lange C, Fezeu L, Jean T, De Lauzon-Guillain B,
Czernichow S, Fumeron F, Froguel P, Vaxillaire M, Cauchi S,
Ducimetière P, Eschwège E.
Predicting diabetes - clinical, biological and genetic approaches: the
D.E.S.I.R. Study.
Diabetes Care, 2008, 31:2056-2061.
Cauchi S, Meyre D, Durand E, Proença C, Marre M, Hadjadj S,
Choquet H, De Graeve F, Gaget S, Allegaert F, Delplanque J,
Permutt MA, Wasson J, Blech I, Charpentier G, Balkau B, Vergnaud
AC, Czernichow S, Patsch W, Chikri M, Glaser B, Sladek R, Froguel P.
Post genome-wide association studies of novel genes associated with
type 2 diabetes show gene-gene interaction and high predictive value.
PLoS ONE, 2008, 3:e2031.
Benzinou M, Chèvre JC, Ward KJ, Lecoeur C, Dina C, Lobbens S,
Durand E, Delplanque J, Horber FF, Balkau B, Borch-Johnsen K,
Hansen T, Pedersen O, Meyre D, Froguel P.
Endocannabinoid receptor 1 gene variations increase risk for obesity and
modulate body mass index in European populations.
Hum Mol Genet, 2008, 17:1916-1921.
Cauchi S, Nead KT, Choquet H, Horber F, Potoczna N, Balkau B,
Marre M, Charpentier G, Froguel P, Meyre D.
The genetic susceptibility to type 2 diabetes may be modulated by obesity
status:implications for association studies.
Benzinou M, Creemers JW, Choquet H, Lobbens S, Dina C,
Durand E, Guerardel A, Boutin P, Jouret B, Heude B, Balkau B,
Tichet J, Marre M, Potoczna N, Horber F, Le Stunff C, Czernichow S,
Sandbaek A, Lauritzen T, Borch-Johnsen K, Andersen G, Kiess W,
Körner A, Kovacs P, Jacobson P, Carlsson LM, Walley AJ, Jørgensen
T, Hansen T, Pedersen O, Meyre D, Froguel P.
Common nonsynonymous variants in PCSK1 confer risk of obesity.
Nat Genet, 2008, 40:943-945.
Chambers Jc, Elliott P, Zabaneh D, Zhang W, Li Y, Froguel P,
Balding D, Scott J, Kooner Js.
Common genetic variation near MC4R is associated with waist
circumference and insulin resistance.
Nat Genet, 2008, 40:716-718.
Blakemore AI, Froguel P.
Is obesity our genetic legacy ?
J Clin Endocrinol Metab, 2008, 93:S51-6.
Dahlman I, Vaxillaire mM Nilsson M, Lecoeur C, Gu HF, CavalcantiProença C, Efendic S, Ostenson CG, Brismar K, Charpentier G,
Gustafsson JA, Froguel P, Dahlman-Wright K, Steffensen KR.
Estrogen receptor alpha gene variants associate with type 2 diabetes and
fasting plasma glucose.
Pharmacogenet Genomics, 2008,18:967-975.
Botton J, Heude B, Maccario J, Ducimetière P, Charles MA. FLVS
Study Group.
Postnatal weight and height growth velocities at different ages between
birth and 5 y and body composition in adolescent boys and girls.
Am J Clin Nutr, 2008, 87:1760-1768.
De Smith AJ, Walters RG, Coin LJ, Steinfeld I, Yakhini Z, Sladek R,
Small deletion variants have stable breakpoints commonly associated
with alu elements.
PLoS ONE, 2008, 3:e3104.
Bouatia-Naji N, De Graeve F, Brönner G, Lecoeur C, Vatin V,
Durand E, Lichtner P, Nguyen TT, Heude B, Weill J, Lévy-Marchal C,
Hebebrand J, Froguel P, Meyre D.
INS VNTR Is Not Associated With Childhood Obesity in 1,023 Families : A
Family-based Study.
Obesity (Silver Spring), 2008,16:1471-1475.
De Smith AJ, Walters RG, Froguel P, Blakemore AI.
Human genes involved incopy number variation : mechanisms of origin,
functional effects and implications for disease.
Cytogenet Genome Res, 2008, 123:17-26.
Duesing K, Charpentier G, Marre M, Tichet J, Hercberg S, Balkau B,
Froguel P, Gibson F.
Evaluating the association of common PBX1 variants with type 2
diabetes.
Bouatia-Naji N, Rocheleau G, Van Lommel L, Lemaire K, Schuit F,
Cavalcanti-Proença C, Marchand M, Hartikainen AL, Sovio U,
De Graeve F, Rung J, Vaxillaire M, Tichet J, Marre M, Balkau B,
Weill J, Elliott P, Jarvelin MR, Meyre D, Polychronakos C, Dina C,
Sladek R, Froguel P.
A Polymorphism Within the G6PC2 Gene Is Associated with Fasting
Plasma Glucose Levels.
Science, 2008, 320:1085-1088.
Duesing K, Fatemifar G, Charpentier G, Marre M, Tichet J,
Hercberg S, Balkau B, Froguel P, Gibson F.
Strong association of common variants in the CDKN2A/CDKN2B region
with type 2 diabetes in French Europids.
Diabetologia, 2008, 51:821-826.
Bouhaha R, Meyre D, Kamoun HA, Ennafaa H, Vaillant E, Sassi R,
Baroudi T, Vatin V, Froguel P, Elgaaied A, Vaxillaire M.
Effect of ENPP1/PC-1-K121Q and PPARgamma-Pro12Ala polymorphisms
on the genetic susceptibility to T2D in the Tunisian population.
Diabetes Res Clin Pract, 2008, 81:278-283.
Duesing K, Fatemifar G, Charpentier G, Marre M, Tichet J,
Hercberg S, Balkau B, Froguel P, Gibson F.
Evaluation of the Association of IGF2BP2 Variants with Type 2 Diabetes
in French Caucasians.
Diabetes, 2008, 57:1992-1996.
Cauchi S, Froguel P.
TCF7L2 genetic defect and type 2 diabetes.
Curr Diab Rep, 2008, 8:149-155.
Flechtner I, Vaxillaire M, Cavé H, Scharfmann R, Froguel P, Polak M.
Neonatal hyperglycaemia and abnormal development of the pancreas.
Best Pract Res Clin Endocrinol Metab, 2008, 22:17-40.
Cauchi S, Proença C, Choquet H, Gaget S, De Graeve F, Marre M,
Balkau B, Tichet J, Meyre D, Vaxillaire M, Froguel P ; D.E.S.I.R. Study
Group.
Analysis of Novel Risk Loci for Type 2 Diabetes in a General French
Population: The D.E.S.I.R. Study.
J Mol Med, 2008, 86:341-348.
The power of the extreme in elucidating obesity.
N Engl J Med, 2008, 359:891-893.
Guan W, Pluzhnikov A, Cox NJ, Boehnke M. International Type 2
Diabetes Linkage Analysis Consortium.
Meta-analysis of 23 type 2 diabetes linkage studies from the International
Type 2 Diabetes Linkage Analysis Consortium.
Hum Hered, 2008, 66:35-49.
Cauchi S, Choquet H, Gutiérrez-Aguilar R, Capel F, Grau K,
Proença C, Dina C,Duval A, Balkau B, Marre M, Potoczna N,
Langin D, Horber F, Sørensen TI, Charpentier G, Meyre D, Froguel P.
Effects of TCF7L2 polymorphisms on obesity in European populations.
137
Contents
Gutiérrez-Aguilar R, Froguel P, Hamid Yh, Benmezroua Y,
Jørgensen T, Borch-Johnsen K, Hansen T, Pedersen O, Neve B.
Genetic analysis of KLF11 variants in 5,864 danish individuals ; potential
effect on insulin resistance and modified stat3 binding by promoter
variant -1659g>C.
Stutzmann F, Tan K, Vatin V, Dina C, Jouret B, Tichet J, Balkau B,
Potoczna N, Horber F, O'rahilly S, Farooqi IS, Froguel P, Meyre D.
Prevalence of MC4R deficiency in European population and their agedependant penetrance in multi-generational pedigrees.
Diabetes, 2008, 57:2511-2518.
Tarasov AI, Nicolson T, Riveline JP, Taneja TK, Baldwin SA,
Baldwin JM, Charpentier G, Gautier JF, Froguel P, Vaxillaire M,
Rutter GA.
A rare mutation in ABCC8/SUR1 leading to altered KATP channel activity
and {beta}-cell glucose sensing is associated with type 2 diabetes mellitus
in adults.
Diabetes, 2008, 57:1595-1604.
Liu YJ, Liu XG, Wang L, Dina C, Yan H, Liu JF, Levy S, Papasian CJ,
Drees BM, Hamilton JJ, Meyre D, Delplanque J, Pei YF, Zhang L,
Recker RR, Froguel P, Deng HW.
Genome-wide association scans identified CTNNBL1 as a novel gene for
obesity.
Hum Mol Genet, 2008, 17:1803-1813.
Martin D, Bellanné-Chantelot C, Deschamps I, Froguel P, Robert Jj,
Velho G.
Long-term follow-up of OGTT-derived glucose tolerance, insulin secretion
and insulin sensitivity indices in subjects with glucokinase mutations
(MODY2).
Diabetes Care, 2008, 31:1321-1323.
Vaxillaire M, Froguel P.
Monogenic diabetes in the young, pharmacogenetics and relevance
to multifactorial forms of type 2 diabetes.
Endocr Rev, 2008, 29:254-264.
Vaxillaire M, Cavalcanti-Proença C, Dechaume A, Tichet J, Marre M,
Balkau B, Froguel P; Desir Study Group.
The common P446L polymorphism in GCKR inversely modulates fasting
glucose and triglyceride levels and reduces type 2 diabetes risk in the
DESIR prospective general French population.
Diabetes, 2008, 57:2253-2257.
Loos RJ, Lindgren CM, Li S, Wheeler E, Zhao JH, Prokopenko I,
Inouye M, Freathy RM, Attwood AP, Beckmann Js, Berndt SJ ; The
Prostate, Lung, Colorectal, And Ovarian (Plco) Cancer Screening
Trial, Jacobs KB, Chanock SJ, Hayes RB, Bergmann S, Bennett AJ,
Bingham SA, Bochud M, Brown M, Cauchi S, Connell JM, Cooper C,
Smith GD, Day I, Dina C, De S, Dermitzakis ET, Doney AS, Elliott KS,
Elliott P, Evans DM, Sadaf Farooqi I, Froguel P, et al.
Common variants near MC4R are associated with fat mass, weight and
risk of obesity.
Nat Genet, 2008, 40:768-775.
Villalobos-Comparán M, Teresa Flores-Dorantes M, Teresa VillarrealMolina M, Rodríguez-Cruz M, García-Ulloa AC, Robles L,
Huertas-Vázquez A, Saucedo-Villarreal N, López-Alarcón M,
Sánchez-Muñoz F, Domínguez-López A, Gutiérrez-Aguilar R,
Menjivar M, Coral-Vázquez R, Hernández-Stengele G, Vital-Reyes Vs,
Acuña-Alonzo V, Romero-Hidalgo S, Ruiz-Gómez Dg, RiañoBarros D, Herrera MF, Gómez-Pérez FJ, Froguel P, García-García E,
Teresa Tusié-Luna M, Aguilar-Salinas CA, Canizales-Quinteros S.
The FTO gene is associated with adulthood obesity in the Mexican
population.
Obesity (Silver Spring), 2008,16:2296-2301.
McAteer JB, Prudente S, Bacci S, Lyon HN, Hirschhorn JN,
Trischitta V, Florez JC. ENPP1 CONSORTIUM.
The ENPP1 K121Q polymorphism is associated with type 2 diabetes in
European populations : evidence from an updated meta-analysis in
42,042 subjects.
Diabetes, 2008, 57:1125-1130.
Wermter AK, Scherag A, Meyre D, Reichwald K, Durand E,
Nguyen TT, Koberwitz K, Lichtner P, Meitinger T, Schäfer H,
Hinney A, Froguel P, Hebebrand J, Brönner G.
Preferential reciprocal transfer of paternal/maternal DLK1 alleles to obese
children : first evidence of polar overdominance in humans.
Eur J Hum Genet, 2008, 16:1126-1134.
Meyre D, Farge M, Lecoeur C, Proenca C, Durand E, Allegaert F,
Tichet J,Marre M, Balkau B, Weill J, Delplanque J, Froguel P.
R125W coding variant in TBC1D1 confers risk for familial obesity and
contributes to linkage on chromosome 4p14 in the French population.
Hum Mol Genet, 2008, 17:1798-1802.
Polak M, Dechaume A, Cavé H, Nimri R, Crosnier H, Sulmont V,
De Kerdanet M, Scharfmann R, Lebenthal Y, Froguel P, Vaxillaire M,
the French ND (Neonatal Diabetes) Group.
Heterozygous Missense Mutations in the Insulin Gene are linked to
Permanent Diabetes appearing in the Neonatal Period or in EarlyInfancy : A report from the French ND Study Group.
Diabetes, 2008, 57:1115-1119.
2009
Poulain-Godefroy O, Lecoeur C, Pattou F, Fruhbeck G, Froguel P.
Inflammation is associated with a decrease of lipogenic factors in
omental fat in women.
Am J Physiol Regul Integr Comp Physiol, 2008, 295:R1-7.
Blakemore AI, Meyre D, Delplanque J, Vatin V, Lecoeur C, Marre M,
Tichet J,Balkau B, Froguel P, Walley AJ.
A Rare Variant in the Visfatin Gene (NAMPT/PBEF1) Is Associated With
Protection From Obesity.
Obesity (Silver Spring), 2009, Mar 19.
Wolowczuk I, Verwaerde C, Viltart O, Delanoye A, Delacre M, Pot B,
Grangette C.
Feeding our immune system : Impact on metabolism.
Clin Develop Immunol, 2008, 1:1-19.
Poulain-Godefroy O, Froguel P.
Response to the letter to the editor : "HIF-1{alpha} protein rather than mRNA
as a marker of hypoxia in adipose tissue in obesity," by Trayhurn et al.
Am J Physiol Regul Integr Comp Physiol, 2008, 295:R1098.
Boissel S, Reish O, Proulx K, Kawagoe-Takaki H, Sedgwick B, Yeo Gs,
Meyre D, Golzio C, Molinari F, Kadhom N, Etchevers Hc, Saudek V,
Farooqi Is, Froguel P, Lindahl T, O'rahilly S, Munnich A, Colleaux L.
Loss-of-function mutation in the dioxygenase-encoding FTO gene causes
severe growth retardation and multiple malformations.
Am J Hum Genet, 2009, 85:106-111.
Roquebert B, Damond F, Collin G, Matheron S, Peytavin G,
Bénard A, Campa P, Chêne G, Brun-Vézinet F, Descamps D; French
Anrs Hiv-2 Cohort (Anrs Co 05 Vih-2).
HIV-2 Integrase Gene Polymorphism and Phenotypic Susceptibility of HIV2 Clinical Isolates to the Integrase Inhibitors Raltegravir and Elvitegravir
in Vitro.
J Antimicrob Chemother, 2008, 62:914-920.
Bonnefond A, Bouatia-Naji N, Simon A, Saint-Martin C,
Dechaume A, De Lonlay P, Polak M, Bellanné-Chantelot C,
Froguel P, Vaxillaire M.
Mutations in G6PC2 do not contribute to monogenic forms of early
infancy diabetes and beta cell dysfunction.
Diabetologia, 2009, 52:982-985.
138
Contents
Bonnefond A, Vaxillaire M, Labrune Y, Lecoeur C, Chèvre Jc, BouatiaNaji N, Cauchi S, Balkau B, Marre M, Tichet J, Riveline JP, Hadjadj S,
Gallois Y, Czernichow S, Hercberg S, Kaakinen M, Wiesner S,
Charpentier G, Lévy-Marchal C, Elliott P, Jarvelin MR, Horber F,
Dina C, Pedersen O, Sladek R, Meyre D, Froguel P.
Genetic variant in HK1 is associated with a proanemic state and A1C but
not other glycemic control-related traits.
Diabetes, 2009, 58:2687-2697.
Chambers JC, Zhang W, Zabaneh D, Sehmi J, Jain P, Mccarthy MI,
Froguel P, Ruokonen A, Balding D, Jarvelin MR, Scott J, Elliott P,
Kooner JS.
Common genetic variation near melatonin receptor MTNR1B contributes
to raised plasma glucose and increased risk of type 2 diabetes among
Indian Asians and European Caucasians.
Diabetes, 2009, 58:2703-2708.
Choquet H, Cavalcanti-Proença C, Lecoeur C, Dina C, Cauchi S,
Vaxillaire M, Hadjadj S, Horber F, Potoczna N, Charpentier G, Ruiz J,
Hercberg S, Maimaitiming S, Roussel R, Boenhnke M, Jackson AU,
Patsch W, Krempler F, Voight BF, Altshuler D, Groop L, Thorleifsson G,
Steinthorsdottir V, Stefansson K, Balkau B, Froguel P, Meyre D.
The T-381C SNP in BNP gene may be modestly associated with type 2
diabetes : an updated meta-analysis in 49 279 subjects.
Hum Mol Genet, 2009, 18:2495-2501.
Bonnet F, Balkau B, Malécot JM, Picard P, Lange C, Fumeron F,
Aubert R, Raverot V, Déchaud H, Tichet J, Lecomte P, Pugeat M ;
Desir Study Group.
Sex hormone-binding globulin predicts the incidence of hyperglycemia
in women : interactions with adiponectin levels.
Eur J Endocrinol, 2009, 161:81-85.
Bossé Y, Bacot F, Montpetit A, Rung J, Qu Hq, Engert JC,
Polychronakos C, Hudson TJ, Froguel P, Sladek R, Desrosiers M.
Identification of susceptibility genes for complex diseases using poolingbased genome-wide association scans.
Hum Genet, 2009, 125:305-318.
Dahlman I, Nilsson M, Gu HF, Lecoeur C, Efendic S, Ostenson CG,
Brismar K, Gustafsson JA, Froguel P, Vaxillaire M, DahlmanWright K, Steffensen KR.
Functional and genetic analysis in type 2 diabetes of liver X receptor
alleles—a cohort study.
Bouatia-Naji N, Bonnefond A, Froguel P.
Inputs from the genetics of fasting glucose : lessons for diabetes.
Med Sci (Paris), 2009, 25:897-902.
De Smith AJ, Purmann C, Walters RG, Ellis RJ, Holder SE,
Van Haelst MM, Brady AF, Fairbrother UL, Dattani M, Keogh JM,
Henning E, Yeo GS, O'rahilly S, Froguel P, Farooqi IS, Blakemore AI.
A Deletion of the HBII-85 Class of Small Nucleolar RNAs (snoRNAs) is
Associated with Hyperphagia, Obesity and Hypogonadism.
Hum Mol Genet, 2009, Jun 4.
Bouatia-Naji N, Bonnefond A, Cavalcanti-Proença C, Sparsø T,
Holmkvist J, Marchand M, Delplanque J, Lobbens S, Rocheleau G,
Durand E, De Graeve F, Chèvre JC, Borch-Johnsen K, Hartikainen AL,
Ruokonen A, Tichet J, Marre M, Weill J, Heude B, Tauber M,
Lemaire K, Schuit F, Elliott P, Jørgensen T, Charpentier G, Hadjadj S,
Cauchi S, Vaxillaire M, Sladek R, Visvikis-Siest S, Balkau B, LévyMarchal C, Pattou F, Meyre D, Blakemore Ai, Jarvelin Mr, Walley AJ,
Hansen T, Dina C, Pedersen O, Froguel P.
A variant near MTNR1B is associated with increased fasting plasma
glucose levels and type 2 diabetes risk.
Nat Genet, 2009, 41:89-94.
Duesing K, Charpentier G, Marre M, Tichet J, Hercberg S, Balkau B,
Froguel P, Gibson F.
Evaluating the association of common APOA2 variants with type 2
diabetes.
El Achhab Y, Meyre D, Bouatia-Naji N, Berraho M, Deweirder M,
Vatin V, Delplanque J, Serhier Z, Lyoussi B, Nejjari C, Froguel P,
Chikri M.
Association of the ENPP1 K121Q polymorphism with type 2 diabetes and
obesity in the Moroccan population.
Diabetes Metab, 2009, 35:37-42.
Bouatia-Naji N, Marchand M, Cavalcanti-Proenca C, Daghmoun S,
Durand E, Tichet J, Marre M, Balkau B, Froguel P, Levy-Marchal C.
Smallness for gestational age interacts with HMGA2 genetic variation to
modulate height.
Eur J Endocrinol, 2009, 160:557-560.
Bouhaha R, Choquet H, Meyre D, Abid Kamoun H, Ennafaa H,
Baroudi T, Sassi R, Vaxillaire M, Elgaaied A, Froguel P, Cauchi S.
TCF7L2 is associated with type 2 diabetes in nonobese individuals from
Tunisia.
Pathol Biol (Paris), 2009, Mar 13.
Elliott P, Chambers Jc, Zhang W, Clarke R, Hopewell JC, Peden Jf,
Erdmann J, Braund P, Engert JC, Bennett D, Coin L, Ashby D,
Tzoulaki I, Brown IJ, Mt-Isa S, Mccarthy MI, Peltonen L, Freimer NB,
Farrall M, Ruokonen A, Hamsten A, Lim N, Froguel P,
Waterworth DM, Vollenweider P, Waeber G, Jarvelin MR, Mooser V,
Scott J, Hall AS, Schunkert H, Anand SS, Collins R, Samani NjJ
Watkins H, Kooner JS.
Genetic Loci associated with C-reactive protein levels and risk of coronary
heart disease.
JAMA, 2009, 302:37-48.
Bouldouyre MA, Charreau I, Marchou B, Tangre P, Katlama C,
Morlat P, Meiffredy V, Vittecoq D, Bierling P, Aboulker JP,
Molina JM ; Anrs 106 Study Group.
Incidence and risk factors of thrombocytopenia in patients receiving
intermittent antiretroviral therapy : a substudy of the ANRS 106-window
trial.
J Acquir Immune Defic Syndr, 2009, 52:531-537.
Ezzidi I, Mtiraoui N, Cauchi S, Vaillant E, Dechaume A, Chaieb M,
Kacem M, Almawi WY, Froguel P, Mahjoub T, Vaxillaire M.
Contribution of type 2 diabetes associated loci in the Arabic population
from Tunisia : a case-control study.
Carlsson LM, Jacobson P, Walley A, Froguel P, Sjöström L, Svensson
PA, Sjöholm K.
ALK7 expression is specific for adipose tissue, reduced in obesity and
correlates to factors implicated in metabolic disease.
Fernandez-Zapico ME, Van Velkinburgh JC, Gutiérrez-Aguilar R,
Neve B, Froguel P, Urrutia R, Stein R.
MODY7 gene, KLF11, is a novel p300-dependent regulator of Pdx-1
(MODY4) transcription in pancreatic islet beta cells.
J Biol Chem, 2009, 284:36482-36490.
Cauchi S, Stutzmann F, Cavalcanti-Proença C, Durand E, Pouta A,
Hartikainen Al, Marre M, Vol S, Tammelin T, Laitinen J, GonzalezIzquierdo A, Blakemore Ai, Elliott P, Meyre D, Balkau B, Järvelin Mr,
Froguel P.
Combined effects of MC4R and FTO common genetic variants on obesity
in European general populations.
J Mol Med, 2009, 87:537-546.
139
Contents
Garcia EA, King P, Sidhu K, Ohgusu H, Walley A, Lecoeur C,
Gueorguiev M, KHalaf S, Davies D, Grossman AB, Kojima M,
Petersenn S, Froguel P, Korbonits M.
The role of ghrelin and ghrelin-receptor gene variants and promoter
activity in type 2 diabetes.
Eur J Endocrinol, 2009, 161:307-315.
Le Roy H, Zuliani T, Wolowczuk I, Faivre N, Jouy N, Masselot B,
Kerkaert JP, Formstecher P, Polakowska R.
Asymetric distribution of epidermal growth factor receptor directs the fate
of normal and cancer keratonicytes.
Stem Cell Develop, 2009, Oct2.
Goossens GH, Petersen L, Blaak EE, Hul G, Arner P, Astrup A,
Froguel P, Patel K, Pedersen O, Polak J, Oppert JM, Martinez JA,
Sørensen TI, Saris WH; Nugenob Consortium.
Several obesity- and nutrient-related gene polymorphisms but not FTO and
UCP variants modulate postabsorptive resting energy expenditure and fatinduced thermogenesis in obese individuals : the NUGENOB study.
Int J Obes (Lond), 2009, 33:669-679.
Limou S, Le Clerc S, Coulonges C, Carpentier W, Dina C, Delaneau O,
Labib T, Taing L, Sladek R, Deveau C, Ratsimandresy R, Montes M,
Spadoni JL, Lelièvre JD, Lévy Y, Therwath A, Schächter F, Matsuda F,
Gut I, Froguel P, Delfraissy JF, Hercberg S, Zagury JF; Anrs Genomic
Group.
Genomewide association study of an AIDS-nonprogression cohort
emphasizes the role played by HLA genes (ANRS Genomewide Association
Study 02).
J Infect Dis, 2009, 199:419-426.
Gueorguiev M, Lecoeur C, Benzinou M, Mein CA, Meyre D, Vatin V,
Weill J, Heude B, Grossman AB, Froguel P, Korbonits M.
A Genetic Study of the Ghrelin and Growth Hormone Secretagogue
Receptor (GHSR) Genes and Stature.
Ann Hum Genet, 2009, 73:1-9.
Lucas S, Verwaerde C, Wolowczuk I.
Is the adipocyte the key road to inflammation ?
Immunol Immunogenetics Insights, 2009, 1:3-14.
Meyre D, Delplanque J, Chèvre Jc, Lecoeur C, Lobbens S, Gallina S,
Durand E, Vatin V, Degraeve F, Proença C, Gaget S, Körner A,
Kovacs P, Kiess W, Tichet J, Marre M, Hartikainen AL, Horber F,
Potoczna N, Hercberg S, Levy-Marchal C, Pattou F, Heude B,
Tauber M, Mccarthy MI, Blakemore AI, Montpetit A, Polychronakos C,
Weill J, Coin LJ, Asher J, Elliott P, Järvelin MR, Visvikis-Siest S,
Balkau B, Sladek R, Balding D, Walley A, Dina C, Froguel P.
Genome-wide association study for early-onset and morbid adult obesity
identifies three new risk loci in European populations.
Nat Genet, 2009, 41:157-159.
Gueorguiev M, Lecoeur C, Meyre D, Benzinou M, Mein CA,
Hinney A, Vatin V, Weill J, Heude B, Hebebrand J, Grossman AB,
Korbonits M, Froguel P.
Association Studies on Ghrelin and Ghrelin Receptor Gene Polymorphisms
With Obesity.
Heid IM, Huth C, Loos RJ, Kronenberg F, Adamkova V, Anand SS,
Ardlie K, Biebermann H, Bjerregaard P, Boeing H, Bouchard C,
Ciullo M, Cooper JA, Corella D, Dina C, Engert JC, Fisher E,
Francès F, Froguel P, et al.
Meta-analysis of the INSIG2 association with obesity including 74,345
individuals : does heterogeneity of estimates relate to study design ?
PLoS Genet, 2009, 5:e1000694.
Morandi A, Pinelli L, Petrone A, Vatin V, Buzzetti R, Froguel P, Meyre D.
The Q121 Variant of ENPP1 May Protect From Childhood
Overweight/obesity in the Italian Population.
Jernås M, Olsson B, Arner P, Jacobson P, Sjöström L, Walley A,
Froguel P, Mcternan PG, Hoffstedt J, Carlsson LM.
Regulation of carboxylesterase 1 (CES1) in human adipose tissue.
Nicolson TJ, Bellomo EA, Wijesekara N, Loder MK, BaldwinM,
Gyulkhandanyan AV, Koshkin V, Tarasov AI, Carzaniga R,
Kronenberger K, Taneja Tk, Da Silva Xavier G, Libert S, Froguel P,
Scharfmann R, Stetsyuk V, Ravassard P, Parker H, Gribble FmM
Reimann F, Sladek R, Hughes SJ, Johnson PR, Masseboeuf M,
Burcelin R, Baldwin SA, Liu M, Lara-Lemus R, Arvan P, Schuit FC,
Wheeler MB, Chimienti F,Rutter GA.
Insulin storage and glucose homeostasis in mice null for the granule zinc
transporter ZnT8 and studies of the type 2 diabetes-associated variants.
Diabetes, 2009, Jun 19.
Juffroy O, Noël D, Delanoye, A, Viltart O, Wolowczuk I, And
Verwaerde C.
Subcutaneous graft of D1 mouse mesenchymal stem cells leads to the
formation of a bone-like structure.
Differentiation, 2009, 78:223-231.
Kamal M, Marquez M, Vauthier V, Leloire A, Froguel P, Jockers R,
Couturier C.
Improved donor/acceptor BRET couples for monitoring beta-arrestin
recruitment to G protein-coupled receptors.
Biotechnol J, 2009, Jun 25.
Prokopenko I, Zeggini E, Hanson RL, Mitchell BD, Rayner NW,
Akan P, Baier L, Das SK, Elliott KS, Fu M, Frayling Tm, Groves CJ,
Gwilliam R, Scott LJ, Voight BF, Hattersley AT, Hu C, Morris AD,
Ng M, Palmer CN, Tello-Ruiz M, Vaxillaire M, Wang CR, Stein L,
Chan J, Jia W, Froguel P, Elbein SC, Deloukas P, Bogardus C,
Shuldiner AR, Mccarthy ML; International Type 2 Diabetes 1q
Consortium.
Linkage disequilibrium mapping of the replicated type 2 diabetes linkage
signal on chromosome 1q.
Diabetes, 2009, 58:1704-1709.
Kong A, Steinthorsdottir V, Masson G, Thorleifsson G, Sulem P,
Besenbacher S, Jonasdottir A, Sigurdsson A, Kristinsson KT,
Jonasdottir A, Frigge ML, Gylfason A, Olason PI, Gudjonsson SA,
Sverrisson S, Stacey SN, Sigurgeirsson B, Benediktsdottir KR,
Sigurdsson H, Jonsson T, Benediktsson R, Olafsson JH, Johannsson OT,
Hreidarsson AB, Sigurdsson G; Diagram Consortium, FergusonSmith AC, Gudbjartsson DF, Thorsteinsdottir U, Stefansson K.
Parental origin of sequence variants associated with complex diseases.
Nature, 2009, 462:868-674.
Rung J, Cauchi S, Albrechtsen A, Shen L, Rocheleau G, CavalcantiProença C, Bacot F, Balkau B, Belisle A, Borch-Johnsen K,
Charpentier G, Dina C, Durand E, Elliott P, Hadjadj S, Järvelin MR,
Laitinen J, Lauritzen T, Marre M, Mazur A, Meyre D, Montpetit A,
Pisinger C, Posner B, Poulsen P, Pouta A, Prentki M, Ribel-Madsen R,
Ruokonen A, Sandbaek A, Serre D, Tichet J, Vaxillaire M,
Wojtaszewski JF, Vaag A, Hansen T, Polychronakos C, Pedersen O,
Froguel P, Sladek R.
Genetic variant near IRS1 is associated with type 2 diabetes, insulin
resistance and hyperinsulinemia.
Nat Genet, 2009, 41:1110-1115.
Le Clerc S, Limou S, Coulonges C, Carpentier W, Dina C, Taing L,
Delaneau O, Labib T, Sladek R; Anrs Genomic Group, Deveau C,
Guillemain H, Ratsimandresy R, Montes M, Spadoni JL, Therwath A,
Schächter F, Matsuda F, Gut I, Lelièvre JD, Lévy Y, Froguel P,
Delfraissy JF, Hercberg S, Zagury JF.
Genomewide association study of a rapid progression cohort identifies new
susceptibility alleles for AIDS (ANRS Genomewide Association Study 03).
J Infect Dis, 2009, 200:1194-1201.
140
Contents
Bacart J, Leloire A, Levoye A, Froguel P, Jockers R, Couturier C.
Evidence For Leptin Receptor Isoforms Heteromerization At The Cell Surface.
Febs Lett, 2010, 584:2213-2217.
Saiki A, Olsson M, Jernås M, Gummesson A, Mcternan Pg,
Andersson J, Jacobson P, Sjöholm K, Olsson B, Yamamura S,
Walley A, Froguel P, Carlsson B, Sjöström L, Svensson PA,
Carlsson LM.
Tenomodulin is highly expressed in adipose tissue, increased in obesity
and down regulated during diet-induced weight loss.
Balkau B, Vol S, Loko S, Andriamboavonjy T, Lantieri O, Gusto G,
Meslier N, Racineux JL, Tichet J.
Epidemiologic Study On The Insulin Resistance Syndrome Study Group.
High Baseline Insulin Levels Associated With 6-Year Incident Observed
Sleep Apnea.
Diabetes Care, 2010, 33(5):1044-1049.
Sparsø T, Bonnefond A, Andersson E, Bouatia-Naji N, Holmkvist J,
Wegner L, Grarup N, Gjesing AP, Banasik K, Cavalcanti-Proença C,
Marchand M, Vaxillaire M, Charpentier G, Jarvelin MR, Tichet J,
Balkau B, Marre M, Lévy-Marchal C, Faerch K, Borch-Johnsen K,
Jørgensen T, Madsbad S, Poulsen P, Vaag A, Dina C, Hansen T,
Pedersen O, Froguel P.
G-allele of intronic rs10830963 in MTNR1B confers increased risk of
impaired fasting glycemia and type 2 diabetes through an impaired
glucose-stimulated insulin release : studies involving 19,605 Europeans.
Diabetes, 2009, 58:1450-1456.
Bonnefond A, Froguel P, Vaxillaire M.
The Emerging Genetics Of Type 2 Diabetes.
Trends Mol Med, 2010, 16:407-416.
Botton J, Heude B, Maccario J, Borys JM, Lommez A, Ducimetière P,
Charles MA ; Flvs Study Group.
Parental Body Size And Early Weight And Height Growth Velocities In Their
Offspring.
Early Hum Dev, 2010,;86 445-450.
Stutzmann F, Cauchi S, Durand E, Calvacanti-Proença C, Pigeyre M,
Hartikainen Al, Sovio U, Tichet J, Marre M, Weill J, Balkau B,
Potoczna N, Laitinen J, Elliott P, Järvelin Mr, Horber F, Meyre D,
Froguel P.
Common genetic variation near MC4R is associated with eating
behaviour patterns in European populations.
Int J Obes (Lond), 2009, 33:373-378.
Bouatia-Naji N, Bonnefond A, Baerenwald DA, Marchand M,
Bugliani M, Marchetti P, Pattou F, Printz RL, Flemming BP,
Umunakwe OC, Conley NL, Vaxillaire M, Lantieri O, Balkau B,
Marre M, Lévy-Marchal C, Elliott P, Jarvelin MR, Meyre D, Dina C,
Oeser JK, Froguel P, O'brien RM.
Genetic And Functional Assessment Of The Role Of The Rs13431652-A
And Rs573225-A Alleles In The G6pc2 Promoter That Strongly Associate
With Elevated Fasting Glucose Levels.
Diabetes, 2010, Jul 9.
Traurig M, Mack J, Hanson RL, Ghoussaini M, Meyre D, Knowler WC,
Kobes S,Froguel P, Bogardus C, Baier LI.
Common variation in SIM1 is reproducibly associated with BMI in Pima
Indians.
Diabetes, 2009, 58:1682-1689.
Bouhaha R, Baroudi T, Ennafaa H, Vaillant E, Abid H, Sassi R, Vatin V,
Froguel P, Gaaied AB, Meyre D, Vaxillaire M.
Study Of Tnfalpha -308g/A And Il6 -174g/C Polymorphisms In Type 2
Diabetes And Obesity Risk In The Tunisian Population.
Clin Biochem, 2010, 43:549-552.
Vaxillaire M, D P, Bonnefond A, Froguel P.
Breakthroughs in monogenic diabetes genetics : from pediatric forms to
young adulthood diabetes.
Pediatr Endocrinol Rev, 2009, 6:405-417.
Cauchi S, Del Guerra S, Choquet H, D'aleo V, Groves CJ, Lupi R,
Mccarthy MI, Froguel P, Marchetti P.
Meta-Analysis And Functional Effects Of The Slc30a8 Rs13266634
Polymorphism On Isolated Human Pancreatic Islets.
Mol Genet Metab, 2010, 100:77-82.
Vierron E, Halimi JM, Tichet J, Balkau B, Cogneau J, Giraudeau B ;
Desir Study Group.
Center effect on ankle-brachial index measurement when using the reference
method (Doppler and manometer) : results from a large cohort study.
Am J Hypertens, 2009, 22:718-722.
Coin LJ, Asher JE, Walters RG, Moustafa JS, De Smith AJ, Sladek R,
Balding DJ, Froguel P, Blakemore AI.
Cnvhap: An Integrative Population And Haplotype-Based Multiplatform
Model Of Snps And Cnvs.
Nat Methods, 2010, 7:541-546.
Vrang N, Meyre D, Froguel P, Jelsing J, Tang-Christensen M, Vatin V,
Mikkelsen JD, Thirstrup K, Larsen LK, Cullberg KB, Fahrenkrug J,
Jacobson P, Sjöström L, Carlsson LM, Liu Y, Liu X, Deng HW,
Larsen PJ.
The Imprinted Gene Neuronatin Is Regulated by Metabolic Status and
Associated With Obesity.
Corpeleijn E, Petersen L, Holst C, Saris WH, Astrup A, Langin D,
Macdonald I, Martinez JA, Oppert JM, Polak J, Pedersen O,
Froguel P, Arner P, Sørensen TI, Blaak EE.
Obesity-Related Polymorphisms And Their Associations With The Ability
To Regulate Fat Oxidation In Obese Europeans : The Nugenob Study.
Walley AJ, Asher JE, Froguel P.
The genetic contribution to non-syndromic human obesity.
Nat Rev Genet, 2009, 10:431-442.
Cuesta-Muñoz AL, Tuomi T, Cobo-Vuilleumier N, Koskela H, Odili S,
Stride A, Buettger C, Otonkoski T, Froguel P, Grimsby J, GarciaGimeno M, Matschinsky FM.
Clinical Heterogeneity In Monogenic Diabetes Caused By Mutations In
The Glucokinase Gene (Gck-Mody).
Diabetes Care, 2010, 33(2):290-292.
Wolowczuk I, Allez M, Chamaillard M.
IL-17/23, potential targets for Crohn’s disease.
Book Chapter in: Progess in Inflammation Research: “Th17 cells : Role
in Inflammation and Autoimmune Disease”, 2009, Chapter : 5, 211-224.
2010
Andersson EA, Holst B, Sparsø T, Grarup N, Banasik K, Holmkvist J,
Jørgensen T, Borch-Johnsen K, Egerod KL, Lauritzen T, Sørensen TI,
Bonnefond A, Meyre D, Froguel P, Schwartz TW, Pedersen O, Hansen T.
Mtnr1b G24e Variant Associates With Bmi And Fasting Plasma Glucose
In The General Population In Studies Of 22,142 Europeans.
Diabetes, 2010, 59:1539-1548.
141
Contents
Dupuis J, Langenberg C, Prokopenko I, Saxena R, Soranzo N,
Jackson AU, Wheeler E, Glazer NL, Bouatia-Naji N, Gloyn AL,
Lindgren Cm, Mägi R, Morris AP, Randall J, Johnson T, Elliott P, Rybin
D, Thorleifsson G, Steinthorsdottir V, Henneman P, Grallert H,
Dehghan A, Hottenga JJ, Franklin CS, Navarro P, Song K, Goel A,
Perry JR, Egan JM, Lajunen T, Grarup N, Sparsø T, Doney A, Voight
BF, Stringham Hm, Li M, Kanoni S, Shrader P, Cavalcanti-Proença C,
Kumari M, Qi L, Timpson NJ, Gieger C, Zabena C, Rocheleau G,
Ingelsson E, An P, O'connell J, Luan J, Elliott A, Mccarroll SA, Payne
F, et al.
New Genetic Loci Implicated In Fasting Glucose Homeostasis And Their
Impact On Type 2 Diabetes Risk.
Nat Genet, 2010, 42:105-116.
Meur G, Simon A, Harun N, Virally M, Dechaume A, Bonnefond A,
Fetita S, Tarasov AI, Guillausseau PJ, Boesgaard TW, Pedersen O,
Hansen T, Polak M, Gautier JF, Froguel P, Rutter GA, Vaxillaire M.
Insulin Gene Mutations Resulting In A Mody Phenotype : Marked
Differences In Clinical Presentation, Metabolic Status And Pathogenic
Effect Through Er Retention.
Diabetes, 2010, 59:653-661.
Meyre D, Proulx K, Kawagoe-Takaki H, Vatin V, Gutiérrez-Aguilar R,
Lyon D, Ma M, Choquet H, Horber F, Van Hul W, Van Gaal L, Balkau B,
Visvikis-Siest S, Pattou F, Farooqi IS, Saudek V, O'rahilly S, Froguel P,
Sedgwick B, Yeo GS.
Prevalence Of Loss Of Function Fto Mutations In Lean And Obese
Individuals.
Diabetes, 2010, 59:311-318.
Freathy RM, Mook-Kanamori DO, Sovio U, Prokopenko I, Timpson NJ,
Berry Dj, Warrington NM, Widen E, Hottenga JJ, Kaakinen M, Lange LA,
Bradfield JP, Kerkhof M, Marsh JA, Mägi R, Chen CM, Lyon HN, Kirin M,
Adair LS, Aulchenko YS, Bennett AJ, Borja JB, Bouatia-Naji N,
Charoen P, Coin LJ, Cousminer DL, De Geus EJ, Deloukas P, Elliott P,
Evans DM, Froguel P; Genetic Investigation Of Anthropometric
Traits (Giant) Consortium, Glaser B, Groves CJ, Hartikainen AL,
Hassanali N, Hirschhorn JN, Hofman A, Holly JM, Hyppönen E,
Kanoni S, Knight BA, Laitinen J, Lindgren CM; Meta-Analyses Of
Glucose And Insulin-Related Traits Consortium, Mcardle WL,
O'reilly PF, Pennell CE, Postma DS, Pouta A, Ramasamy A, Rayner NW,
Ring SM, Rivadeneira F, Shields BM, Strachan DP, Surakka I, Taanila A,
Tiesler C, Uitterlinden AG, Van Duijn CM; Wellcome Trust Case
Control Consortium, Wijga AH, Willemsen G, Zhang H, Zhao J,
Wilson JF, Steegers EA, Hattersley AT, Eriksson JG, Peltonen L,
Mohlke KL, Grant SF, Hakonarson H, Koppelman GH, Dedoussis GV,
Heinrich J, Gillman MW, Palmer LJ, Frayling TM, Boomsma DI, Davey
Smith G, Power C, Jaddoe VW, Jarvelin MR; Early Growth Genetics
(Egg) Consortium, Mccarthy MI.
Variants In Adcy5 And Near Ccnl1 Are Associated With Fetal Growth And
Birth Weight.
Nat Genet, 2010, 42:430-435.
Miot A, Maimaitiming S, Emery N, Bellili N, Roussel R, Tichet J, Velho G,
Balkau B, Marre M, Fumeron F; Desir Study Group.
Genetic Variability At The Six Transmembrane Protein Of Prostate 2 Locus
And The Metabolic Syndrome : The Data From An Epidemiological Study
On The Insulin Resistance Syndrome (Desir) Study.
J Clin Endocrinol Metab. 2010 Jun;95(6):2942-7.
Morandi A, Maffeis C, Lobbens S, Bouatia-Naji N, Heude B, Pinelli L,
Meyre D, Froguel P.
Early Detrimental Metabolic Outcomes Of Rs17300539-A Allele Of Adipoq
Gene Despite Higher Adiponectinemia.
Olsson M, Olsson B, Jacobson P, Thelle DS, Björkegren J, Walley A,
Froguel P, Carlsson LM, Sjöholm K.
Expression Of The Selenoprotein S (Sels) Gene In Subcutaneous Adipose
Tissue And Sels Genotype Are Associated With Metabolic Risk Factors.
Metabolism, 2010, Jul 8.
Poulain-Godefroy O, Le Bacquer O, Plancq P, Lecoeur C, Pattou F,
Frühbeck G, Froguel P.
Inflammatory Role Of Toll-Like Receptors In Human And Murine Adipose
Tissue.
Mediators Inflamm, 2010, 2010:823486.
Ghoussaini M, Stutzmann F, Couturier C, Vatin V, Durand E,
Lecoeur C, Degraeve F, Heude B, Tauber M, Hercberg S, LevyMarchal C, Tounian P, Weill J, Traurig M, Bogardus C, Baier LJ,
Michaud JL, Froguel P, Meyre D.
Analysis Of The Sim1 Contribution To Polygenic Obesity In The French
Population.
Repapi E, Sayers I, Wain LV, Burton PR, Johnson T, Obeidat M,
Zhao JH, Ramasamy A, Zhai G, Vitart V, Huffman JE, Igl W, Albrecht E,
Deloukas P, Henderson J, Granell R, Mcardle WL, Rudnicka AR;
Wellcome Trust Case Control Consortium, Barroso I, Loos RJ,
Wareham NJ, Mustelin L, Rantanen T, Surakka I, Imboden M,
Wichmann HE, Grkovic I, Jankovic S, Zgaga L, Hartikainen AL,
Peltonen L, Gyllensten U, Johansson A, Zaboli G, Campbell H,
Wild SH, Wilson JF, Gläser S, Homuth G, Völzke H, Mangino M,
Soranzo N, Spector TD, Polasek O, Rudan I, Wright AF, Heliövaara M,
Ripatti S, Pouta A, Naluai AT, Olin AC, Torén K, Cooper MN,
James AL, Palmer LJ, Hingorani AD, Wannamethee SG, Whincup PH,
Smith GD, Ebrahim S, Mckeever TM, Pavord ID, Macleod AK,
Morris AD, Porteous DJ, Cooper C, Dennison E, Shaheen S,
Karrasch S, Schnabel E, Schulz H, Grallert H, Bouatia-Naji N,
Delplanque J, Froguel P, Blakey JD; Nshd Respiratory Study Team,
Britton JR, Morris RW, Holloway JW, Lawlor DA, Hui J, Nyberg F,
Jarvelin MR, Jackson C, Kähönen M, Kaprio J, Probst-Hensch NM,
Koch B, Hayward C, Evans Dm, Elliott P, Strachan DP, Hall Ip,
Tobin MD.
Genome-Wide Association Study Identifies Five Loci Associated With Lung
Function.
Nat Genet, 2010, 42:36-44.
Grau K, Cauchi S, Holst C, Astrup A, Martinez JA, Saris WH, Blaak EE,
Oppert JM, Arner P, Rössner S, Macdonald Aa, Klimcakova E,
Langin D, Pedersen O, Froguel P, Sørensen TI.
Tcf7l2 Rs7903146-Macronutrient Interaction In Obese Individuals'
Responses To A 10-Wk Randomized Hypoenergetic Diet.
Am J Clin Nutr, 2010, 91:472-479.
Ma L, Hanson RL, Traurig MT, Muller YL, Kaur BP, Perez JM, Meyre D,
Fu M, Körner A, Franks PW, Kiess W, Kobes S, Knowler WC, Kovacs P,
Froguel P, Shuldiner AR, Bogardus C, Baier LJ.
Evaluation Of A2bp1 As An Obesity Gene.
Diabetes, 2010, Aug 19.
Maimaitiming S, Roussel R, Hadjadj S, Fumeron F, Aubert R,
Emery N, Velho G, Mohammedi K, Travert F, Tichet J, AlhencGelas F, Balkau B, Marre M; D.E.S.I.R.
Study Group. Association Of Common Variants In Nppa And Nppb With
Blood Pressure Does Not Translate Into Kidney Damage In A General
Population Study.
J Hypertens, 2010, 28:1230-1233.
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Saxena R, Hivert MF, Langenberg C, Tanaka T, Pankow JS,
Vollenweider P, Lyssenko V, Bouatia-Naji N, Dupuis J, Jackson AU,
Kao WH, Li M, Glazer NL, Manning AK, Luan J, Stringham HM,
Prokopenko I, Johnson T, Grarup N, Boesgaard TW, Lecoeur C,
Shrader P, O'connell J, Ingelsson E, Couper DJ, Rice K, Song K,
Andreasen CH, Dina C, Köttgen A, Le Bacquer O, Pattou F,
Taneera J, Steinthorsdottir V, Rybin D, Ardlie K, Sampson M, Qi L,
Van Hoek M, Weedon MN, Aulchenko YS, Voight BF, Grallert H,
Balkau B, Bergman RN, Bielinski SJ, Bonnefond A, Bonnycastle LL,
Borch-Johnsen K, et al.
Genetic Variation In Gipr Influences The Glucose And Insulin Responses
To An Oral Glucose Challenge.
Nat Genet, 2010, 42:142-148.
Walters RG, Jacquemont S, Valsesia A, De Smith AJ, Martinet D,
Andersson J, Falchi M, Chen F, Andrieux J, Lobbens S, Delobel B,
Stutzmann F, El-Sayed Moustafa JS, Chèvre JC, Lecoeur C, Vatin V,
Bouquillon S, Buxton JL, Boute O, Holder-Espinasse M, Cuisset JM,
Lemaitre MP, Ambresin AE, Brioschi A, Gaillard M, Giusti V,
Fellmann F, Ferrarini A, Hadjikhani N, Campion D, Guilmatre A,
Goldenberg A, Calmels N, Mandel JL, Le Caignec C, David A,
Isidor B, Cordier MP, Dupuis-Girod S, Labalme A, Sanlaville D,
Béri-Dexheimer M, Jonveaux P, Leheup B, Ounap K, Bochukova EG,
Henning E, Keogh J, Ellis RJ, Macdermot KD, Van Haelst MM,
Vincent-Delorme C, Plessis G, Touraine R, Philippe A, Malan V,
Mathieu-Dramard M, Chiesa J, Blaumeiser B, Kooy RF, Caiazzo R,
Pigeyre M, Balkau B, Sladek R, Bergmann S, Mooser V, Waterworth D,
Reymond A, Vollenweider P, Waeber G, Kurg A, Palta P, Esko T,
Metspalu A, Nelis M, Elliott P, Hartikainen AL, Mccarthy MI, Peltonen L,
Carlsson L, Jacobson P, Sjöström L, Huang N, Hurles Me, O'rahilly S,
Farooqi Is, Männik K, Jarvelin MR, Pattou F, Meyre D, Walley AJ,
Coin LJ, Blakemore AI, Froguel P, Beckmann JS.
A New Highly Penetrant Form Of Obesity Due To Deletions On
Chromosome 16p11.2.
Nature, 2010, 463:67167-5.
Scherag A, Dina C, Hinney A, Vatin V, Scherag S, Vogel CI, Müller TD,
Grallert H, Wichmann HE, Balkau B, Heude B, Jarvelin MR,
Hartikainen AL, Levy-Marchal C, Weill J, Delplanque J, Körner A,
Kiess W, Kovacs P, Rayner NW, Prokopenko I, Mccarthy MI,
Schäfer H, Jarick I, Boeing H, Fisher E, Reinehr T, Heinrich J,
Rzehak P, Berdel D, Borte M, Biebermann H, Krude H, Rosskopf D,
Rimmbach C, Rief W, Fromme T, Klingenspor M, Schürmann A,
Schulz N, Nöthen MM, Mühleisen TW, Erbel R, Jöckel KH, Moebus S,
Boes T, Illig T, Froguel P, Hebebrand J, Meyre D.
Two New Loci For Body-Weight Regulation Identified In A Joint Analysis
Of Genome-Wide Association Studies For Early-Onset Extreme Obesity In
French And German Study Groups.
Plos Genet, 2010, 6:E1000916.
Yamauchi T, Hara K, Maeda S, Yasuda K, Takahashi A, Horikoshi M,
Nakamura M, Fujita H, Grarup N, Cauchi S, Ng DP, Ma RC,
Tsunoda T, Kubo M, Watada H, Maegawa H, Okada-Iwabu M,
Iwabu M, Shojima N, Shin HD, Andersen G, Witte DR, Jørgensen T,
Lauritzen T, Sandbæk A, Hansen T, Ohshige T, Omori S, Saito I,
Kaku K, Hirose H, So WY, Beury D, Chan JC, Park KS, Tai ES, Ito C,
Tanaka Y, Kashiwagi A, Kawamori R, Kasuga M, Froguel P,
Pedersen O, Kamatani N, Nakamura Y, Kadowaki T.
A Genome-Wide Association Study In The Japanese Population Identifies
Susceptibility Loci For Type 2 Diabetes At Ube2e2 And C2cd4a-C2cd4b.
Nat Genet, 2010, Sep 5.
Su SY, Asher JE, Jarvelin MR, Froguel P, Blakemore AI, Balding DJ,
Coin LJ.
Inferring Combined Cnv/Snp Haplotypes From Genotype Data.
Bioinformatics, 2010, 26:1437-445.
Vasseur F, Caeyseele T, Barat-Houari M, Lobbens S, Meirhaeghe A,
Meyre D, Froguel P, Amouyel P, Helbecque N.
Concordance Of Two Multiple Analytical Approaches Demonstrate That
Interaction Between Bmi And Adipoq Haplotypes Is A Determinant Of Ldl
Cholesterol In A General French Population.
J Hum Genet, 2010, 55:227-231.
PhD
Fanny STUTZMANN
Directeur de thèse : Philippe FROGUEL
Sujet de thèse : Le récepteur 4 aux mélanocortines, un paradigme de la
génétique de l'obésité
9 Septembre 2009
Voight BF, Scott LJ, Steinthorsdottir V, Morris AP, Dina C, Welch RP,
Zeggini E, Huth C, Aulchenko YS, Thorleifsson G, Mcculloch LJ,
Ferreira T, Grallert H, Amin N, Wu G, Willer CJ, Raychaudhuri S,
Mccarroll SA, Langenberg C, Hofmann OM, Dupuis J, Qi L, Segrè AV,
Van Hoek M, Navarro P, Ardlie K, Balkau B, Benediktsson R,
Bennett AjJ Blagieva R, Boerwinkle E, Bonnycastle LL, Bengtsson
Boström K, Bravenboer B, Bumpstead S, Burtt NP, Charpentier G,
Chines PS, Cornelis M, Couper DJ, Crawford G, Doney AS, Elliott KS,
Elliott AL, Erdos MR, Fox CS, Franklin CS, Ganser M, Gieger C,
Grarup N, Green T, Griffin S, Groves CJ, Guiducci C, Hadjadj S,
Hassanali N, Herder C, Isomaa B, Jackson AU, Johnson PR,
Jørgensen T, Kao WH, Klopp N, Kong A, Kraft P, Kuusisto J,
Lauritzen T, Li M, Lieverse A, Lindgren CM, et al.
Twelve Type 2 Diabetes Susceptibility Loci Identified Through Large-Scale
Association Analysis.
Nat Genet, 2010, 42:579-589.
HDR
David Meyre
Sujet de thèse : Identification de variants génétiques de prédisposition à
l’obésité
16 Février 2009
Vrang N, Meyre D, Froguel P, Jelsing J, Tang-Christensen M, Vatin V,
Mikkelsen JD, Thirstrup K, Larsen LK, Cullberg KB, Fahrenkrug J,
Jacobson P, Sjöström L, Carlsson LM, Liu Y, Liu X, Deng HW,
Larsen PJ.
The Imprinted Gene Neuronatin Is Regulated By Metabolic Status And
Associated With Obesity.
Cyril Couturier
Sujet de thèse : les récepteurs à la leptine et leur protéine associée,
interactions et implications dans l’obesité.
9 octobre 2009
143
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144
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Nuclear Receptors,
Cardiovascular Diseases
and Diabetes
U 1011 Inserm
affiliated to IFR 114 & IFR 142
Bart STAELS
Contact :
00 33 3 20 87 78 25
[email protected]
Besin Isabelle, Administrative staff NSFA
Tournay Sylvie, Administrative staff Lille 2
Chartier François, Administrative Responsable IPL
Derudas Marie-Hélène, Administrative staff IPL
Frémaux Jacques, Technician IPL
Lebel Pascal, Assistant IPL
Lefebvre Claudine, Administrative staff IPL
Libouraux Chantal, Administrative staff INSERM
Martin-Nizard Françoise, MCU Lille 2
Charles Evelyne, Technician IPL
Duthoit Béatrice, Technician Lille 2
Watrain Annick, Technician IPL
145
Contents
4 Teams
• Nuclear receptors in the metabolic syndrome
Bart STAELS
• Molecular control of monocyte / macrophage functions
in cardiometabolic syndrome
Giulia CHINETTI
• Immuno-inflammation and cardiovascular diseases
David DOMBROWICZ
• Molecular analysis of gene regulation in cardiometabolic diseases
Philippe LEFEBVRE
146
Contents
of synthetic reference as well as novel ligands allowed to study
the biological effects of these compounds. These studies
might uncover novel therapeutic approaches for the
treatment of the metabolic abnormalities predisposing to
atherosclerosis, such as type 2 diabetes, dyslipidemia or
obesity. Within this context, team 3 demonstrated that PPARa
improves the adaptative response of the pancreatic beta-cell
to pathological conditions and showed a direct role for FXR
(Farnesoid X Receptor) in the regulation of peripheral insulin
sensitivity, adipocyte function and energy homeostasis.
Perturbations of the circadian rhythm are known to increase
the incidence of metabolic diseases. Team 3 is studying the
implication of nuclear receptors in the clock system and has
contributed to the identification of the nuclear receptor Reverba as an integrator of circadian and metabolic systems. The
objective of team 4 was to identify and study the regulation
of human macrophage genes involved in foam cell formation.
The identification of genes involved in lipid accumulation
within the arterial wall or implicated in cellular redox systems,
two important pathways contributing to arterial pathology
and its complications, contributed to a better understanding
of the role of macrophages in the development of
atherosclerosis.
A - Research Report
During the present 4-year term (2006-2009), we have been
studying the molecular mechanisms involved in the
development of atherosclerosis, with a particular focus on the
role and mechanism of action of nuclear receptors, their
involvement in the pathological processes, and their potential
usefulness as therapeutic targets.
The Unit 545 is composed of four teams studying convergent
and complementary research topics :
Team 1 : Clinical research and experimental pharmacology
G. Luc
Team 2 : Nuclear receptors and the vascular wall
B. Staels
Team 3 : Systemic risk factors of atherosclerosis : regulation
and modulation by nuclear receptors
C. Fiévet
Team 4 : Metabolic syndrome, arterial inflammation and
rupture of atheroma plaques
M. Rouis (who left the laboratory in November 2007).
As a participant in large consortia, team 1 has access to
important epidemiological cohorts, such as the Prospective
Epidemiological Study of Myocardial Infarction (PRIME Study),
on which a variety of biological parameters are measured,
such as inflammatory and angiogenesis factors, proteins
involved in the metabolism of lipoproteins, hormones,
adipokines, metalloproteinases, and protease inhibitors.
Identification of novel cardiovascular risk factors provides
opportunities to investigate their molecular action
mechanism and leads to the identification of new genes
involved in cardiovascular diseases. Other teams of the Unit
study their in vivo role using original pathophysiological
animal models and develop collaborations with clinical teams.
The regulation of newly identified genes, as well as the
metabolic consequences of their modulation, is also studied.
Team 2 focuses on the study of the molecular mechanisms
controlling vascular lipid homeostasis, inflammation in
macrophages as well as atherosclerosis progression. A specific
emphasis is put on the functions of two nuclear receptor
subfamilies, namely the PPARs (Peroxisome ProliferatorActivated Receptors) and LXRs (Liver X Receptors). Team 2
showed that these nuclear receptor subfamilies regulate
cholesterol trafficking to the plasma membrane thus reducing
intracellular cholesteryl ester accumulation. Moreover, LXR
ligands increase the expression of the LPS-receptor Toll like
receptor (TLR)-4 in human macrophages, as well as ROS
generation by enhancing the expression of NADPH oxidase
subunits, pinpointing a role of LXR as an integrator of inflammatory signaling. Finally, team 2 has recently demonstrated
that human atherosclerotic plaques contain not only
classically activated M1 macrophages which produce proinflammatory cytokines, but also M2 macrophages, which
have anti-inflammatory properties, and that activation of
PPARg primes primary human monocytes both in vitro and in
vivo, into M2 differentiation. To obtain further insights into the
role of nuclear receptors in metabolic regulation, team 3
devoted most of its activities to development and study of
pathophysiological animal models. Moreover, since nuclear
receptors are ligand-regulated transcription factors, the use
development 2010
We will study the biological and molecular mechanisms
controlling the development of atherosclerosis in order to
develop preventive and therapeutic strategies against its
cardiovascular complications, and study metabolic risk factors,
such as dyslipidemia and type 2 diabetes. We will focus on the
regulation of genes involved in these pathologies and on the
consequences of their dysregulation, studying the molecular
basis and the pathophysiological involvement of nuclear
receptors, transcription factors which our Unit has been
studying for several years. Four different teams will be
organized around a Unit project entitled “Nuclear receptors,
cardiovascular diseases and diabetes”. Team 1 (Nuclear
receptors in the metabolic syndrome) will study the metabolic
147
Contents
functions of nuclear receptors (FXR, Rev-erbα, RORα) and their
interest as potential therapeutic targets in humans. Team 2
(Molecular control of monocyte/macrophage functions in the
cardiometabolic syndrome) will focus its activities on the role
of PPARg in peripheral blood mononuclear cells from obese
subjects, and will study the role of PPARs, LXRs and the cyclindependent kinase inhibitor CDKN2Ap16INK4a in the molecular
regulation of monocyte differentiation and macrophage
functions in the vascular wall and adipose tissues during
aobesity. The mechanisms by which nuclear receptors control
the contribution of other cells from the immune system to
atherosclerosis will be studied by team 3 (Immunoinflammation and cardiovascular diseases) with a particular
interest for the control by PPARγ of mast cell and B lymphocyte
functions. Team 4 (Molecular analysis of gene regulation in
cardiometabolic diseases) will investigate how nuclear
receptors control gene expression and affect metabolic
functions, allowing the unraveling of novel regulatory
pathways and the validation of nuclear receptors as
pharmacological targets. Our Unit possesses all required
expertise in cellular and molecular biological techniques, and
benefits from comprehensive technical platforms in
biochemistry and histology. The Unit has also a longstanding
experience in the development and study of new animals
models in particular genetically-modified mice.
148
Contents
identified FXR as a modulator of energy homeostasis. We have
clearly shown that FXR plays a direct role in the regulation of
peripheral insulin sensitivity and in adipocyte function. We
also identified two new FXR target genes.
Finally, we have studied the biological functions of Rev-erbα
and RORα, two nuclear receptors which cross-talk with other
nuclear receptors, i.e. with LXR in the macrophage, and link
circadian signals and metabolic processes. We thus
demonstrated that Rev-erbα positively regulates hepatic bile
acid synthesis in vivo by de-repressing CYP7A1 gene
repression via SHP and E4BP4 transcriptional repression.
We have shown that enterocytes express certain of these
nuclear receptors and may be interesting pharmacological
targets in the control of cholesterol homeostasis.
Nuclear Receptors in the
Metabolic Syndrome
Bart Staels
PU Lille 2
Group Members :
Bailleul Bernard, DR2 Inserm
Duez Helene, CR1 Inserm
Lestavel Sophie, PU Lille 2
Briand Olivier, MCU Lille 2
Caron-Houde Sandrine, MCU Lille 2
Lalloyer Fanny, MCU Lille 2
Baugé Eric, ITA IPL
Dorchies Emilie, ITA IPL
Duhem Christian, ITA IPL
Lucas Anthony, ITA Inserm
Touche Véronique, ITA Lille 2
Daoudi Mehdi, Postdoctoral fellow
Prawitt Janne, Postdoctoral fellow
Sebti Yasmine, Postdoctoral fellow
Woldt Estelle, Postdoctoral fellow
Colin Sophie, PhD Student Lille 2
Huaman Samanez Carolina, PhD Student Lille 2
Trabelsi Sami, Master 2 Student Lille 2
development 2010
The general objective of this research project is to better
understand the metabolic functions of nuclear receptors, with
a major focus on FXR, Rev-erbα and RORα the cross-talk
pathways with other nuclear receptors, and to define the
potential benefit of pharmacological agents acting via these
receptors on human health.
To obtain further insights in the regulatory role of these nuclear
receptors in metabolism and energy homeostasis, total- or
organ-specific deficient or over-expressing animal models of
the nuclear receptors of interest are already available or in
development. The impact of these nuclear receptors will be
investigated by comparing them to wild-type mice with respect
to basal metabolic parameters, energy expenditure, gene and
protein expression and pharmacological response.
Concomitantly, molecular approaches will be applied to identify
the molecular mechanisms at the basis of the identified
physiological functions using various in vitro models (cell lines
or primary cells in culture) and cell and molecular biology
approaches (transfection, quantitative PCR, Western-blot,
Chromatin Immunoprecipitation (ChIP), gene silencing, DNA
micro-array technology, ChIP-seq technology…). Since nuclear
receptors are potential pharmacological targets, the use of
existing as well as the identification of novel synthetic ligands
will allow us to study the biological effects of these compounds.
Finally, the role of these nuclear receptors in human physiology
will be investigated by analysis of tissue biopsies from subjects
suffering from metabolic disorders, obtained through our active
participation in different European and national consortiums.
It is expected that these approaches will uncover new
therapeutic strategies for the treatment of metabolic diseases
such as type 2 diabetes and dyslipidemia.
Key words :
Metabolism. Genetically modified organism. Molecular
and cellular pharmacology. Nuclear receptors. Pathophysiology
A - Research Report
The organism senses its energy status through the close
communication of several organs (liver, adipose tissue,
intestine, pancreas), which integrate multiple endocrine
(hormones, inflammatory cytokines) and metabolic (glucose,
free fatty acids and intermediary metabolites) signals.
Dysregulation of the tight control of metabolism leads to
dyslipidemia, insulin resistance and obesity which predispose
to the development of cardiovascular complications due to
atherosclerosis. Nuclear receptors play key roles in the
(patho)physiological adaptation to alterations in energy
status.
During the last four years, we focused our research on the role
of the fatty acid and fibrate-activated PPARs in whole-body
metabolism and deciphered the molecular mechanisms of
their anti-atherosclerotic activities through its triglyceride
decreasing and HDL and reverse cholesterol transport
enhancing activities. Moreover, PPARα improves the adaptative
response of the pancreatic beta-cell to pathological conditions.
Using molecular approaches, we were able to define the
selective PPAR modulator (SPPARM) concept.
We also explored the role of bile acid-receptor FXR in the
regulation of hepatic and peripheral glucose metabolism and
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Contents
Cousein E, Barthelemy C, Poullain S, Simon N, Lestavel S,
Williame V, Joiris E, Danel C, Clavey V, Brossard D, Robert H, CrausteManciet S, Vaccher C, Odou P.
P-glycoprotein and cytochrome P450 3A4 involvement in risperidone
transport using an in vitro Caco-2/TC7 model and an in vivo model.
Prog Neuro-psych Biol Psych, 2007, 4 :878-886.
Drouineaud V, Sagot P, Garrido C, Logette E, Deckert V, Gambert P,
Jimenez C, Staels B, Lagrost L, Masson D.
Inhibition of progesterone production in human luteinized granulosa cells
treated with LXR agonists.
Mol Hum Reprod, 2007, 13:73-379.
Falcetti E, Flavell DM, Staels B, Tinker A, Haworth SG, Clapp LH.
IP receptor-dependent activation of PPARg by stable prostacyclin analogues.
Fievet C, Fruchart JC, Staels B.
Genetically-engineered animals as research models for atherosclerosis :
their use for the characterization of PPAR agonists in the treatment of
cardiometabolic disorders.
Front Biosci, 2007, 12:4132-4156.
Publications
Flajollet S, Lefebvre B, Cudejko C, Staels B, Lefebvre P.
The core component of the mammalian SWI/SNF complex
SMARCD3/BAF60c is a coactivator for the nuclear retinoic acid receptor.
Mol Cell Endocrinol, 2007, 270:3-32.
2007
Balmforth AJ, Grant PJ, Scott EM, Wheatcroft SB, Kearney MT,
Staels B, Marx N.
Inter-subject differences in constitutive expression levels of the clock gene
in man.
Diab Vasc Dis Res, 2007, 40 :39-43.
Fontaine C, Staels B.
The orphan nuclear receptor Rev-erb alpha : a transcriptional link
between circadian rhythmicity and cardiometabolic disease.
Curr Opin Lipidol, 2007, 18:141-146.
Barter P, Gotto AM, Larosa JC, Maroni J, Szarek M, Grundy SM,
Kastelein JJ, Bittner V, Fruchart JC For The Treating To New Targets
Investigators.
HDL cholesterol, very low levels of LDL cholesterol, and cardiovascular events.
N Engl J Med, 2007, 357:1301-1310.
Fruchart JC.
Is lipoprotein(a) a clinically meaningful risk marker for cardiovascular
events in healthy women ?
Nat Clin Pract Cardiovasc Med, 2007, 4:242-243.
Blanc-Brude OP, Teissier E, Castier Y, Leseche G, Bijnens AP,
Daemen M, Staels B, Mallat Z, Tedgui A.
IAP survivin regulates atherosclerotic macrophage survival.
Fruchart JC.
Novel peroxisome proliferator activated receptor-alpha agonists.
Am J Cardiol, 2007, 100:S41-S46.
Gervois P, Fruchart JC, Staels B.
Drug insight : mechanisms of action and therapeutic applications for
agonists of PPAR.
Nat Clin Pract Endocrinol Metab, 2007, 3:145-156.
Brigadeau F, Gele P, Wibaux M, Marquie C, Martin-Nizard F,
Torpier G, Fruchart JC, Staels B, Duriez P, Lacroix D.
The PPARa activator fenofibrate slows down the progression of the left
ventricular dysfunction in porcine tachycardia-induced cardiomyopathy.
J Cardiovasc Pharmacol, 2007, 49:408-415.
Gross B, Staels B.
PPAR agonists : multimodal drugs for the treatment of type-2 diabetes.
Best Pract Res Clin Endoc Met, 2007, 21:687-710.
Cariou B, Bouchaert E, Abdelkarim M, Dumont J, Caron S,
Fruchart JC, Burcelin R, Kuipers F, Staels B.
FXR-deficiency confers increased susceptibility to torpor.
FEBS Letters, 2007, 581:5191-5198.
Hondares E, Pineda-Torra I, Iglesias R, Staels B, Villarroya F, Giralt M.
PPARdelta, but not PPARalpha, activates PGC-1a gene transcription in muscle.
Cariou B, Staels B.
FXR : a promising target for the metabolic syndrome ?
Trends Pharmacol Sci, 2007, 28:36-243.
Kahri J, Fruchart-Najib J, Matikainen N, Fruchart JC, Vakkilainen J,
Taskinen MR.
The increase of apolipoprotein A-V during postprandial lipemia parallels
the response of triglyceride-rich lipoproteins in type 2 diabetes: no
relationship between apo A-V and postheparin plasma lipolytic activity.
Diabetes Care, 2007, 30:2083-2085.
Carmona MC, Louche K, Lefebvre B, Pilon A, Hennuyer N, AudinotBouchez V, Fievet C, Torpier G, Formstecher P, Renard P, Lefebvre P,
Dacquet C, Staels B, Casteilla L, Penicaud L on behalf of the
Consortium of the French Ministry of Research and Technology.
S 26948 : a new specific PPARg modulator with potent antidiabetes and
antiatherogenic effects.
Diabetes, 2007, 56 : 2797-2808.
Kuipers F, Stroeve JHM, Caron S, Staels B.
Bile acids, farnesoid X receptor, atherosclerosis and metabolic control.
Curr Opin Lipidol, 2007, 18:289-297.
Chinetti-Gbaguidi G, Staels B.
Measuring biomarkers to assess the therapeutic effects of PPAR agonists ?
Pharmacogenomics, 2007, 8:1567-1580.
Murakami T, Walczak R, Caron S, Duhem C, Vidal V, Darteil R, Staels B.
The farnesoid X receptor induces fetuin-B gene expression in human
hepatocytes.
Biochem J, 2007, 407:461-469.
150
Contents
Nakamachi T, Nomiyama T, Gizard F, Heywood EB, Jones KI, Zhao Y,
Fuentes L, Takebayashi K, Aso Y, Staels B, Inukai T, Bruemmer D.
PPARa agonists suppress osteopontin expression in macrophages and
decrease plasma levels in patients with type 2 diabetes.
Diabetes, 2007, 56 :1662-1670.
Bultel S, Helin L, Clavey V, Chinetti-Gbaguidi G, Rigamonti E,
Colin M, Fruchart JC, Staels B, Lestavel S.
Liver X receptor activation induces the uptake of cholesteryl esters from
high density lipoproteins in primary human macrophages.
Niculescu LS, Fruchart-Najib J, Fruchart JC, Sima A.
Apolipoprotein A-V gene polymorphisms in subjects with metabolic
syndrome.
Clin Chem Lab Med, 2007, 45:1133-1139.
Calpe-Berdiel L, Rotllan N, Fievet C, Roig R, Blanco-Vaca F, EscolaGil JC.
Liver X Receptor-mediated activation of reverse cholesterol transport from
macrophages to feces in vivo requires ABCG5/G8.0
J Lipid Res, 2008, 49:1904-1911.
Paumelle R, Staels B.
PPAR mediate pleiotropic actions of statins.
Circ Res, 2007, 100:394-1395.
Caron S, Staels B.
Apolipoprotein C-III : A link between hypertriglyceridemia and vascular
dysfunction ? (Editorial).
Circ Res, 2008, 103:1348-1350.
Quemeneur T, Martin-Nizard F, Kandoussi A, Kyndt X, Vanhille P,
Haculla E, Hatron PY, Fruchart JC, Duriez P, Lambert M.
PON1, a new biomarker of cardiovascular disease, is low in patients with
systemic vasculitis.
Semin Arthritis Rheum, 2007, 37:149-155.
Carpentier R, Sacchetti P, Segard P, Staels B, Lefebvre P.
The glucocorticoid receptor is a co-regulator of the orphan nuclear
receptor Nurr1.
J Neurochem, 2008, 104:777-789.
Raichur S, Lau P, Staels B, Muscat GE.
Retinoid-related orphan receptor gamma regulates several genes that
control metabolism in skeletal muscle cells : links to modulation of
reactive oxygen species production.
J Mol Endocrinol, 2007, 39:29-44.
Clemenz M, Frost N, Schupp M, Caron S, Foryst-Ludwig A, Bohm C,
Hartge M, Gust R, Staels B, Unger T, Kintscher U.
Liver-specific PPARa-target gene regulation by the angiotensin type 1
receptor blocker telmisartan.
Diabetes, 2008, 57:1405-1413.
Rubenstrunk A, Hanf R, Hum DW, Fruchart JC, Staels B.
Safety issues and prospects for future generations of PPAR modulators.
Colin S, Bourguignon E, Boullay AB, Tousaint JJ, Huet S, Caira F,
Staels B, Lestavel S, Lobaccaro JM, Delerive P.
Intestine-specific regulation of PPARalpha gene transcription by Liver X
Receptors.
Endocrinology, 2008, 149:5128-5135.
Sharma AM, Staels B
PPARgamma and adipose tissue – Understanding obesity-related
changes in regulation of lipid and glucose metabolism.
Denechaud PD, Bossard P, Lobaccaro JMA, Millatt L, Staels B,
Girard J, Postic C.
ChREBP, but not LXRs, is required for the induction of glucose-regulated
genes in mouse liver.
J Clin Invest, 2008, 118:956-964.
Staels B.
PPAR agonists and the metabolic syndrome.
Thérapie, 2007, 62:319-326.
Duez H, Lamarche B, Uffelman KD, Valéro R, Szeto L, Lemieux S,
Cohn JS, Lewis GF.
Dissociation between the insulin sensitizing effect of rosiglitazone and
its effect on hepatic and intestinal lipoprotein production.
Staels B, Kuipers F.
Bile acid sequestrants and the treatment of type 2 diabetes mellitus.
Drugs, 2007, 67:1383-1392.
Telliez A, Desroses M, Pommery N, Briand O, Farce A, Laconde G,
Lemoine A, Depreux P, Henichart JP.
Derivatives of Iressa, a specific epidermal growth factor receptor inhibitor,
are powerful apoptosis inducers in PC3 prostatic cancer cells.
Chem Med Chem, 2007, 2/3:318-332.
Duez H, Lamarche B, Valéro R, Pavlic M, Proctor S, Xiao C, Szeto L,
Patterson BW, Lewis GF.
Both intestinal and hepatic lipoprotein production are stimulated by an
acute elevation of plasma free fatty acids in humans.
Circulation, 2008, 117 :2369-2376.
Yamazaki Y, Abe K, Toma T, Nishikawa M, Ozawa H, Okuda A,
Araki T, Oda S, Inoue K, Shibuya K., Staels B, Fruchart JC.
Design and synthesis of highly potent and selective human peroxisome
proliferator-activated receptor α agonists. Bioorg Med
Chem Lett, 2007, 17:4689-4693.
Duez H, Pavlic M, Lewis GF.
Mechanism of intestinal lipoprotein overproduction in insulin resistant humans.
Atherosclerosis Suppl, 2008, 9:33-38.
Duez H, Staels B.
The nuclear receptors Rev-erbs and RORs integrate circadian rhythms and
metabolism.
Diabetes Vasc Dis Res, 2008, 5 :82-88.
Zawadzki C, Susen S, Richard F, Haulon S, Corseaux D, Jeanpierre E,
Vincentelli A, Lucas C, Torpier G, Martin A, Van Belle E, Staels B, Jude B.
Dyslipidemia shifts the tissue factor/tissue factor pathway inhibitor
balance toward increased thrombogenicity in atherosclerotic plaques.
Evidence for a corrective effect of statins.
Duez H, Staels B.
Rev-erba gives a time cue to metabolism.
FEBS Lett, 2008, 582:19-25.
2008
Duez H, Van Der Veen J, Duhem C, Pourcet B, Touvier T, Fontaine C,
Derudas B, Bauge E, Havinga R, Bloks VW, Wolters H, Van Der
Sluijs FH, Vennstrom B, Kuipers F, Staels B.
Regulation of bile acid synthesis by the orphan nuclear receptor Reverbalpha.
Bouhlel A, Staels B, Chinetti-Gbagudi G.
Glitazones in the modulation of macrophage lipid homeostasis and the
inflammatory response.
Athero Org Commentaries, International Atherosclerosis Society
Website, July 2008.
151
Contents
Fan Y, Wang Y, Tang Z, Zhang H, Qin X, Zhu Y, Guan Y, Wang X,
Staels B, Chien S, Wang N.
Suppression of pro-inflammatory adhesion molecules by PPAR-β/δ in
human vascular endothelial cells.
Mansouri RM, Bauge E, Gervois P, Fruchart-Najib J, Fievet C,
Staels B, Fruchart JC.
Atheroprotective effect of human apolipoprotein A5 in a mouse model
of mixed dyslipidemia.
Circ Res, 2008, 103:450-453.
Finck BN, Chinetti G, Staels B.
Editorial : PPARs/RXRs in cardiovascular physiology and disease.
PPAR Res, 2008, 173780.
Mansouri R, Bauge E, Staels B, Gervois P.
Systemic and distal repercussions of liver-specific PPARa control of the
acute phase response.
Endocrinology, 2008, 149:3215-3223.
Fontaine C, Rigamonti E, Pourcet B, Duez H, Fruchart JC, ChinettiGbaguidi G, Staels B.
The orphan nuclear receptor Rev-erbalpha is a novel Liver X Receptor
(LXR) target gene driving a negative feedback loop on select LXR-induced
pathways in human macrophages.
Mol Endocrinology, 2008, 22:1797-1811.
Parmenon C, Guillard J, Caignard DH, Hennuyer N, Staels B,
Audinot-Bouchez V, Boutin JA, Dacquet C, Ktorza A, ViaudMassuard MC.
4,4-dimethyl-1,2,3,4-tetrahydroquinoline-based PPARalpha/gamma
agonists. Part I: synthesis and pharmacological evaluation.
Bioorg Med Chem Lett, 2008, 18:1617-1622.
Fruchart JC, Sacks FM, Hermans MP, Assmann G, Brown WV, Ceska R,
Chapman MJ, Dodson P., Fioretto P, Ginsberg HN, Kadowaki T,
Lablanche JM, Marx N, Plutzki J, Reiner Z, Rosenson RS, Staels B,
Stock JK, Sy R, Wanner C, Zambon A, Zimmet P for the Residual Risk
Reduction Initiative.
The residual risk reduction initiative : a call to action to reduce residual
vascular risk in patients with dyslipidemia.
Am J Cardiol, 2008, 102:Suppl. 1K-34K.
Cross-talk between statins and PPARalpha in cardiovascular diseases :
clinical evidence and basic mechanisms.
Trends Cardiovasc Med, 2008, 18:73-78.
Pavlic M, Valero R, Duez H, Xiao C, Szeto L, Patterson BW, Lewis GF.
Triglyceride-rich lipoprotein-associated apolipoprotein CIII production is
stimulated by plasma free fatty acids in humans.
Gineste R, Sirvent A, Paumelle R, Helleboid S, Aquilina A, Darteil R,
Hum DW, Fruchart JC, Staels B.
Phosphorylation of farnesoid X receptor by protein kinase C promotes its
transcriptional activity.
Mol Endocrinol, 2008, 22:2433-2447.
Rigamonti E, Chinetti-Gbaguidi G, Staels B.
Regulation of macrophage functions by PPARalpha, PPARgamma, and
LXRs in mice and men.
Gizard F, Nomiyama T, Zhao Y, Findeisen HM, Heywood EB,
Jones KL, Staels B, Bruemmer D.
The PPARα/p16INK4a pathway inhibits vascular smooth muscle cell
proliferation by repressing cell cycle-dependent telomerase activation.
Circ Res, 2008, 103:1155-1163.
Rigamonti E, Fontaine C, Lefebvre B, Duhem C, Lefebvre P, Marx N,
Staels B, Chinetti-Gbaguidi G.
Induction of CXCR2 receptor by PPARg in human macrophages.
Jelassi A, Najah M, Jguirim I, Maatouk F, Lestavel S, Laroussi OS,
Rouis M, Boileau C, Rabes JP, Varret M, Slimane MN.
A novel splice site mutation of the LDL receptor gene in a Tunisian
hypercholesterolemic family.
Clin Chim Acta, 2008, 392:25-29.
Simoncic M, Resen T, Juvan P, Fievet C, Staels B, Rozman D, Horvat S.
Transcriptome analysis revealed association of some P450 genes with
obesity in a polygenic obese mouse model.
Acta Chim Slov, 2008, 55:101-110.
Kaeding J, Bouchaert E, Belanger J, Caron P, Chouinard S, Verreault M,
Larouche O, Pelletier G, Staels B, Belanger A, Barbier O.
Activators of the farnesoid X receptor negatively regulate androgen
glucuronidation in human prostate cancer LNCAP cells.
Biochem J, 2008, 410:245-253.
Smeets PJH, Teunissen BEJ, Willemsen PHM, Van Nieuwenhoven FA,
Brouns AE, Janssen BJA, Cleutjens JPM, Staels B, Van Der Vusse GJ,
Van Bilsen M.
Cardiac hypertrophy is enhanced in PPARalpha-/- mice in response to
chronic pressure-overload.
Cardiovasc Res, 2008, 78:79-89.
Langhi C, Le May C, Kourimate S, Caron S, Staels B, Krempf M,
Costet P, Cariou B.
Activation of the farnesoid X receptor represses PCSK9 expression in
human hepatocytes.
FEBS Lett, 2008, 582:949-955.
Staels B, Maes M, Zambon A.
Fibrates and future PPARα agonists in the treatment of cardiovascular
disease.
Nat Clin Pract Cardiovasc Med, 2008, 5:542-553.
Legry V, Cottel D, Ferrieres J, Chinetti G, Deroide T, Staels B,
Amouyel P, Meirhaeghe A.
Association between liver X receptor alpha gene polymorphisms and risk
of metabolic syndrome in French populations.
Int J Obes, 2008, 32 :421-428.
Staumont-Salle D, Abboud G, Brenuchon C, Kanda A, Roumier T,
Lavogiez C, Fleury S, Remy P, Papin JP, Bertrand-Michel J, Terce F,
Staels B, Delaporte E, Capron M, Dombrowicz D.
PPARalpha regulates skin inflammation and humoral response in atopic
dermatitis.
MacDonald ML, Singaraja RR, Bissada N, Ruddle P, Watts R,
Karasinska JM, Gibson WT, Fievet C, Vance JE, Staels B, Hayden MR.
Absence of stearoyl-CoA desaturase-1 ameliorates features of the
metabolic syndrome in LDLR-deficient mice.
J Lipid Res, 2008, 49:217-229.
Tancevski I, Wehinger A, Demetz E, Eller P, Duwensee K, Huber J,
Hochegger K, Schgoer W, Fievet C, Stellaard F, Rudling M, Patsch JR,
Ritsch A.
Reduced plasma HDL cholesterol in hyperthyroid mice coincides with
decreased hepatic adenosine 5’-triphosphate-binding cassette
transporter 1 expression.
Endocrinology, 2008, 149:3708-3712.
152
Contents
Dol-Gleizes F, Paumelle R, Visentin V, Mares AM, Desitter P,
Hennuyer N, Gilde A, Staels B, Schaeffer P, Bono F.
Rimonabant, a selective cannabinoid CB1 receptor antagonist, inhibits
atherosclerosis in LDL receptor-deficient mice.
Wouters K, Van Gorp PJ, Bieghs V, Gijbels MJ, Duimel H, Lutjohann D,
Kerksiek A, Van Kruchten R, Maeda N, Staels B, Van Bilsen M,
Shiri-Sverdlov R, Hofker MH.
Dietary cholesterol, rather than liver steatosis, leads to hepatic inflammation
in hyperlipidemic mouse models of non-alcoholic steatohepatitis.
Hepatology, 2008, 48:474-486.
Duez H, Duhem C, Laitinen S, Patole PS, Abdelkarim M, BoisJoyeux B, Danan JL, Staels B.
Inhibition of adipocyte differentiation by RORα.
FEBS Letters, 2009, 583:2031-2036.
Zamble A, Martin-Nizard F, Sahpaz S, Hennebelle T, Staels B,
Bordet R, Duriez P, Brunet C, Bailleul F.
Vasoactivity, antioxidant and aphrodisiac properties of Caesalpinia
benthamiana roots.
J Ethnopharmacol, 2008, 116 :112-119.
Duez H, Smith A, Xiao, Giacca A, Szeto L, Drucker DJ, Lewis GF.
Acute DPP-4 Inhibition rapidly enhances insulin-mediated suppression of
endogenous glucose production in mice.
Endocrinol, 2009, 150:59-62.
2009
Bougarne N, Paumelle R, Caron S, Hennuyer N, Mansouri R, Gervois
P, Staels B, Haegeman G, De Bosscher K.
PPARα blocks glucocorticoid receptor α-mediated transactivation but
cooperates with the activated glucocorticoid receptor α for
transrepression on NF-κB.
Duez H, Staels B.
Rev-erbα: an integrator of circadian rhythms and metabolism.
J Appl Physiol, 2009, 107 :1972-1980.
Fievet C, Staels B.
Efficacy of PPAR agonists in diabetes and coronary artery disease.
Curr Atheroscler Rep, 2009, 11:281-288.
Bougarne N, Paumelle R, Haegeman G, Staels B, De Bosscher K.
Circumventing glucocorticoid-mediated hyperinsulinemia via the
activation of PPARα.
Cell Cycle, 2009, 8:2311-2312.
Fievet C, Staels B.
Liver X Receptor modulators : effects on lipid metabolism and potential
use in the treatment of atherosclerosis.
Biochem Pharmacol, 2009, 77:1316-1327.
Bouhlel MA, Brozek J, Derudas B, Zawadzki C, Jude B, Staels B,
Chinetti-Gbaguidi G.
Unlike PPARγ, PPARα or PPARβ/δ activation does not promote human
monocyte differentiation toward alternative macrophages.
Fievet C, Staels B.
Combination therapy of statins and fibrates in the management of
cardiovascular risk.
Curr Opinion Lipidol, 2009, 20:505-511.
Brinster S, Lamberet G, Staels B, Trieu-Cuot P, Gruss A, Poyart C.
Type II fatty acid synthesis is not a suitable antibiotic target for Grampositive pathogens.
Nature, 2009, 458:83-86.
Giugliano D, Standl E, Vilsboll T, Betteridge J, Bonadonna R,
Campbell IW, Schernthaner GH, Staels B, Trichopoulou A, Farinaro E.
Is the current therapeutic armamentarium in diabetes enough to control
the epidemic and its consequences ? What are the current shortcomings ?
Acta Diabetol, 2009, 46:173-181.
Brunham LR, Singaraja RR, Duong M, Timmins JM, Fievet C,
Bissada N, Kang MH, Samra A, Fruchart JC, Mcmanus B, Staels B,
Parks JS, Hayden MR.
Tissue-specific roles of ABCA1 influence susceptibility to atherosclerosis.
Goebel M, Clemenz M, Staels B, Unger T, Kintscher U, Gust R.
Characterization of new PPAR-gamma agonists : analysis of Telmisartan’s
structural components.
Chem Med Chem, 2009, 4:445-456.
Carreon-Torres E, Rendon-Sauer K, Monter-Garrido M, ToledoIbelles P, Gamboa R, Menjivar M, Lopez-Marure R, Luc G, Fievet C,
Cruz D, Vargas-Alarcon G, Perez-Mendez O.
Rosiglitazone modifies HDL structure and increases HDL-apo AI synthesis
and catabolic rate.
Clin Chim Acta, 2009, 401:37-41.
Goebel M, Staels B, Unger T, Kintscher U, Gust R.
Characterization of new PPARγ agonists : benzimidazole derivatives – the
importance of position 2.
Chem Med Chem, 2009, 4:1136-1142.
Charton J, Deprez-Poulain R, Hennuyer N, Tailleux A, Staels B, Deprez B.
Novel non-carboxylic acid retinoids : 1,2,4-oxadiazol-5-one derivatives.
Bioorg Med Chem Lett, 2009, 19/2:489-492.
Jeanpierre E, Le Tourneau T, Zawadzki C, Van Belle E, Mouquet F,
Susen S, Ezekowitz MD, Staels B, Jude B, Corseaux D.
Beneficial effects of fenofibrate on plaque thrombogenicity and plaque
stability in atherosclerotic rabbits.
Cardiovasc Pathol, 2009, 18:140-147.
Lipid ligand-activated transcription factors regulating lipid storage and
release in human macrophages.
Lalloyer F, Pedersen TA, Gross B, Lestavel S, Yous S, Vallez E,.
Gustafsson JA, Mandrup S, Fievet C, Staels B, Tailleux A.
Rexinoid bexarotene modulates triglyceride but not cholesterol metabolism
via gene-specific permissivity of the RXR/LXR heterodimer in the liver.
van Dijk TH, Grefhorst A, Oosterveer MH, Bloks VQ, Staels B,
Reijngoud DJ, Kuipers F.
An increased flux through the glucose 6-phosphate pool in enterocytes
delays glucose absorption in Fxr-/- mice.
J Biol Chem, 2009, 284/10315-10323.
Lefebvre P, Cariou B, Lien F, Kuipers F, Staels B.
Role of bile acids and bile acid receptors in metabolic regulation.
Physiol Rev, 2009, 89:147-191.
153
Contents
Macdonald ML, Van Eck M, Hildebrand RB, Wong BW, Bissada N,
Ruddle P, Kontush A, Hussein H, Pouladi MA, Chapman MJ, Fievet
C, Van Berkel TJ, Staels B, Mcmanus BM, Hayden MR.
Despite antiatherogenic metabolic characteristics, SCD1-deficient mice
have increased inflammation and atherosclerosis.
Bauge E, Rouille Y, Salles JP, Staels B, Bailleul B.
action in mice.
J Clin Invest, 2009, 119:3830-3838.
Zamble A, Martin-Nizard F, Sahpaz S, Reynaert ML, Staels B,
Bordet R, Duriez P, Gressier B, Bailleul F.
Effects of microdesmis keayana alkaloids on vascular parameters of
erectile dysfunction.
Phytother Res, 2009, 23:892-895.
Mysiorek C, Culot M, Dehouck L, Derudas B, Staels B, Bordet R,
Cecchelli R, Fenart L, Berezowski V.
Peroxisome-proliferator-activated receptorα activation protects brain
capillary endothelial cells from oxygen-glucose deprivation-induced
hyperpermeability in the blood-brain barrier.
Curr Neurovasc Res, 2009, 6:181-193.
Zawadzki C, Chatelain N, Delestre M, Susen S, Quesnel F, Juthier F,
Jeanpierre E, Azzaoui R, Corseaux D, Breyne J, Torpier G, Staels B,
Van Velle E, Jude B.
Tissue factor pathway inhibitor-2 gene methylation is associated with
low expression in carotid atherosclerotic plaques.
Ogata M, Tsujita M, Hossain MA, Akita N, Gonzalez FJ, Staels B,
Suzuki S, Fukutomi T, Kimura G, Yokoyama S.
On the mechanism for PPAR agonists to enhance ABCA1 gene expression.
Oosterveer MH, Grefhorst A, Van Dijk TH, Havinga R, Staels B,
Kuipers F, Groen AK, Reijngoud DJ.
Fenofibrate simultaneously induces hepatic fatty acid oxidation,
synthesis and elongation in mice.
J Biol Chem, 2009, 284 :34036-34044.
2010
Brinster S, Lamberet G, Staels B, Trieu-Cuot P, Gruss A, Poyart C.
Efficient FASH-inhibition does not block staphylococcal growth in fattyacid-rich medium.
Nature (Brief Communications Arising), 2010, 463:E3-E5.
Parmenon C, Guillard J, Caignard DH, Hennuyer N, Staels B, AudinotBouchez V, Boutin JA, Dacquet C, Ktorza A, Viaud-Massuard MC.
agonists. Part II : synthesis and pharmacological evaluation of oxime and
acidic head group structural variations.
Duez H, Staels B.
Nuclear receptors linking circadian rhythms and cardiometabolic control.
Arterioscler. Thromb. Vasc. Biol., 2010, 30, 1529-1534.
Fonseca VA, Handelsman Y, Staels.
Colesevelam lowers glucose and lipid levels in type 2 diabetes : the clinical
evidence.
Diabetes Obes Metab, 2010, 12:384-392.
Porta N, Vallee L, Lecointe C, Bouchaert E, Staels B, Bordet R, Auvin S.
Fenofibrate, a peroxisome proliferator-activated receptor-α agonist,
exerts anticonvulsive properties.
Epilepsia, 2009, 50:943-948.
Goebel M, Wolber G, Markt P, Staels B, Unger T, Kintscher U, Gust R.
Characterization of new PPARγ agonists : benzimidazole derivatesimportance of positions 5 and 6, and computational studies on the
binding mode.
Bioorg Med Chem, 2010, 18:5885-5895.
Provost AC, Vede L, Bigot K, Keller N, Tailleux A, Jais JP, Savoldelli M,
Ameqrane I, Lacassagne E, Legeais JM, Staels B, Menasche M,
Mallat Z, Behar-Cohen F, Abitbol M.
Morphologic and electroretinographic phenotype of SR-BI knockout mice
after a long-term atherogenic diet.
Guillaumond F, Grechez-Cassiau A, Subramaniam M, Brangolo S,
Peteri-Brunback B, Staels B, Fievet C, Spelsberg TC, Delaunay F,
Teboul M.
Krüppel-like factor KLF10 is a link between the circadian clock and
metabolism in liver.
Mol Cell Biol, 2010, 30:3059-3070.
Rambaud J, Triqueneaux G, Masse I, Staels B, Laudet V, Benoit G.
Rev-erbα2 mRNA encodes a stable protein with a potential role in
circadian clock regulation.
Mol Endocrinol, 2009, 23:630-639.
Lalloyer F, Staels B.
Fibrates, Glitazones, and Peroxisome Proliferator-Activated Receptors.
Rost TH, Haugan Moi LL, Berge K, Staels B, Mellgren G, Berge RK.
A pan-PPAR ligand induces hepatic fatty acid oxidation in PPARalpha-/mice possibly through PGC-1 mediated PPARdelta coactivation.
Lauressergues E, Staels B, Valeille K, Majd Z, Hum DH, Duriez P,
Cussac D.
Antipsychotic drug action on SREBPs-related lipogenesis and
cholesterogenesis in primary rat hepatocytes.
Naunyn-Schmied.
Arch Pharmacol, 2010, 381:427-439.
Staels B.
A review of bile acid sequestrants : potential mechanism(s) for glucoselowering effects in type 2 diabetes mellitus.
Postgrad Med, 2009, 121:Suppl 1, 25-30.
SUMOylation of the human peroxisome proliferator-activated receptorα
inhibits trans-activity through the recruitment of the nuclear corepressor
NCoR.
J Biol Chem, 2010, 285:5983-5992.
Staels B, Fonseca VA.
Bile acids and metabolic regulation. Mechanisms and clinical responses
to bile acid sequestration.
Diabetes Care, 2009, 32:S237-S245.
Tancevski I, Wehinger A, Demetz E, Hoefer J, Eller P, Huber E, Stanzl
U, Duwensee K, Auer K, Schgoer W, Kuhn V, Fievet C, Stellaard F,
Rudling M, Foeger B, Patsch JR, Ritsch A.
The thyromimetic T-0681 protects from atherosclerosis.
J Lipid Res, 2009, 50:938-944.
Prawitt J, Beil FT, Marshall RP, Bartelt A, Ruether W, Heeren J,
Amling M, Staels B, Niemeier A.
Short-term activation of liver X receptors inhibits osteoblasts but longterm activation does not have an impact on murine bone in vivo.
Bone, 2010, in press
154
Contents
Riahi Y, Sin-Malia Y, Cohen G, Alpert E, Gruzman A, Eckel J, Staels B,
Guichardant S. Sasson S.
The natural protective mechanism against hyperglycemia in vascular
endothelial cells. Roles of the lipid peroxidation product 4hydroxydodecadienal and PPARδ.
Diabetes, 2010, 59:808-818.
Thierry TOUVIER
Directeur de thèse : Bernard Bailleul
« Les Tétraspanes leprot et leprotL1 : 2 nouveaux régulateurs négatifs de
la sensibilité cellulaire à l'hormone de croissance »
02 février 2009
Ristea Popescu I, Helleboid-Chapman A, Lucas A, Vandewalle B,
Dumont J, Bouchaert E, Derudas B, Kerr-Conte J, Caron S, Pattou F,
Staels B.
The nuclear receptor FXR is expressed in pancreatic β-cells and protects
human islets from lipotoxicity.
FEBS Letters, 2010, 584:2845-2851.
Iuliana RISTEA
Directeur de thèse : Bart Staels
« Rôle du récepteur nucléaire FXR dans le métabolisme du glucose : étude
de son expression, régulation et fonction dans le pancréas endocrine »
14 décembre 2009
Staels B.
Fibrates in CVD : a step towards personalised medicine.
Lancet, 2010, 375:1847-1848.
Mouaadh ABDELKARIM
Directeur de thèse : Catherine Fiévet
Rôle du récepteur nucléaire Farnesoid X Receptor (FXR) dans la
différenciation et la fonction adipocytaire.
12 mai 2010
Staels B.
Introduction on the ATVB Review Series “Nuclear receptors in metabolism
and cardiovascular disease”.
HDR
Staels B, Handelsman Y, Fonseca V.
Bile acid sequestrants for lipid and glucose control.
Curr Diab Rep. 2010, 10:70-77.
Françoise MARTIN-NIZARD
8 décembre 2009
Stienstra R, Saudale F, Duval C, Keshtkar S, Groener JEM,
Van Rooijen N, Staels B, Kersten S, Muller M.
Kupffer cells promote hepatic steatosis via interleukin-1β-dependent
suppression of PPARα activity.
Hepatology, 2010, 51:511-522.
Stroeve JH, Brufau G, Stellaard F, Gonzalez FJ, Staels B, Kuipers F.
Intestinal FXR-mediated FGF15 production contributes to diurnal control
of hepatic bile acid synthesis in mice.
Lab Invest, 2010, in press.
Wouters K, Van Bilsen V, Van Gorp PJ, Bieghs V, Lutjohann D,
Kerksiek A, Staels B, Hofker MH, Shiri-Sverdlov R.
Intrahepatic cholesterol influences progression, inhibition and reversal
of non-alcoholic steatohepatitis in hyperlipidemic mice.
FEBS Lett, 2010, 584:1001-1005.
PhD
Loredane NICULESCU
Directeur de thèse : Patrick Duriez
« Les mécanismes de l’action de l’apolipoprotéine apo AV dans le
métabolisme des lipides »
Co-tutelle Université de Lille 2/Académie Roumaine de Bucarest
18 juin 2007
Lionel HELIN
Directeur de thèse : Véronique Clavey
« Régulation de l’homéostasie du cholestérol par les récepteurs nucléaires
PPARα et LXR dans les macrophages primaires humains »
14 décembre 2007
Romain GINESTE
Directeur de thèse : Jean-Charles Fruchart
« Régulation de l'activité transcriptionnelle des récepteurs nucléaires FXR,
RORalpha et PPARgamma par modification post-traductionnelle »
18 novembre 2008
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Contents
display anti-inflammatory effects involving PPARα in
macrophages. Finally, using the apolipoprotein E2-knock-in
mouse model (apoE2KI), we have evaluated the effects of
PPARα, PPARγ and RXR agonists on atherogenesis.
Molecular Control of
Monocyte / Macrophage
Functions in Cardiometabolic Syndrome
Giulia CHINETTI
DR2 Inserm
Group Members :
Hennuyer Nathalie, Research assistant, IPL
Lestrelin-Paumelle Réjane, PU Lille 2
Muhr-Tailleux Anne, PU Lille 2
Copin Corinne, ITA IPL
Derudas Bruno, ITA IPL
D’Huysser Aurore, ITA IPL
Dubanchet Barbara, ITA Lille 2
Duplan Isabelle, ITA Lille 2
Lepage Joëlle, ITA Inserm
Paquet Charlotte, ITA Lille 2
Vallez Emmanuelle, ITA IPL
Vanhoutte Jonathan, ITA IPL
Verney Aurore, ITA IPL
Blum Kadiombo, MCU Lille 2
Wouters Kristiaan, Postdoctoral fellow
Baron Morgane, PhD Student Lille 2
Bories Gaël, PhD Student Lille 2
Cudejko Céline, PhD Student Lille 2
Mayi Thérèse, PhD Student Lille 2
development 2010
Monocyte derived-macrophages are central cells in the
development of metabolic disorders and related diseases such
as obesity and atherosclerosis, since they can infiltrate adipose
tissue as well as the vascular wall, respectively, contributing
to the maintenance of an inflammatory status. However, two
different sub-populations of macrophages have been
identified in adipose tissue and atherosclerotic lesions: proinflammatory “classical” M1 and anti-inflammatory
“alternative” M2 macro-phages, and their ratio is suggested to
play a role in the metabolic complications of obesity. On the
other hand, it is thought that adipose tissue products may, in
turn, modulate peripheral blood mononuclear cells (PBMC)
and monocyte functions.
During the past few years we focused our investigations on
the effects of PPAR and LXR activation on circulating
monocyte-derived macrophages, with particular emphasis on
their role on modulation of lipid homeostasis and
inflammation. We have also explored the role of PPARα in
smooth muscle cell proliferation and identified
CDKN2A(p16INK4a) as a cognate target gene.
Key words :
Cellular biology. Molecular biology. Molecular and cellular
pharmacology. Cardiovascular disease. Obesity.
A - Research Report
Since the macrophage plays an important role in host defence
and immuno-inflammatory pathologies, our group has been
studying, the last four years, the role of PPARs and LXRs in the
the control of lipid homeostasis and inflammation in
macrophages as well as in atherosclerosis progression. In
particular, we have shown that activation of both PPARα and
LXRs regulate intracellular cholesterol trafficking to the plasma
membrane, hence reducing intracellular cholesteryl ester
accumulation. Moreover, we have shown that LXR ligands
increase the expression of the LPS receptor TLR (Toll like
receptor)-4 in human macrophages, as well as ROS generation
by enhancing the expression of NADPH oxidase subunits, thus
suggesting a role of LXRs in the modulation of the human
macrophage response against bacteria. In addition, we have
demonstrated that human atherosclerotic plaques contain
both classically activated M1 macrophages which produce
pro-inflammatory cytokines, and alternative M2 macrophages
which have anti-inflammatory properties, and that activation
of PPARγ primes primary human monocytes, both in vitro and
in vivo, into the M2 phenotype. During the last years we have
also been interested by the pharmacological modulation of
macrophage functions by statins, a class of hypocholesterolemiant molecules, and have shown that these drugs
Our general objective is to better define the molecular
functions by nuclear receptors (PPARs and LXRs) and
CDKN2A(p16INK4a) and to determine the consequences of
their cross-talk in the context of cardiometabolic diseases.
More precisely, we will study the impact of obesity and its
comorbidities on monocyte functions and their modulation
by PPARg. We will also analyze the molecular mechanisms
controlling the differentiation of monocytes into M1 and
M2 macrophages and the role of PPARs, LXRs and
CDKN2A(p16INK4a) in this process. Finally, we will study the
functions of these nuclear receptor and CDKN2A(p16INK4a)
in macrophage tissues involved in cardiometabolic diseases
such as adipose tissues and atherosclerotic plaques.
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Contents
To get further inside into the role of these factors in regulation
of monocyte/macrophage functions, human and/or mouse
materials (peripheral blood mononuclear cells, bone marrow
derived macrophages, macrophage in adipose tissue and
atherosclerotic plaques) will be analyzed. Animal models with
total deficiency of the factor of interest as well as bone marrow
transplantation technology will be adopted to reinforce our
project. Laser capture micro-dissection, histology, cellular
biology and biochemical approaches will be used.
Concomitantly, molecular biology methods (quantitive PCR,
western blot, DNA-microarray and ChIP technologies) will be
combined to delineate the molecular mechanisms at the basis
of the identified functions.
Fontaine C, Rigamonti E, Nohara A, Gervois P, Teissier E, Fruchart JC,
Liver X receptor activation potentiates the lipopolysaccharide response
in human macrophages.
Circ Res, 2007, 101:40-49.
Publications
PPAR mediate pleiotropic actions of statins.
Circ Res, 2007, 100:394-1395.
Larigauderie G, Jaye M, Rouis M.
Towards the elucidation of the role of perilipin in human macrophages.
Nicaud V, Francomme C, Ruidavets JB, Luc G, Arveiler D, Kee F, Evans
A, Morrison C, Blankenberg S, Cambien F, Tiret L.
Lack of association between complement factor H polymorphisms and
coronary artery disease or myocardial infarction.
J Mol Med, 2007, 85:771-775.
2007
Perez-Mendez O, Alvarez-Salcedo P, Carreon-Torres E, Luc G, Arce
Fonseca M, De la Pena A, Cruz Robles D, Garcia JJ, Vargas-Alarcon G.
Palmitic acid in HDL is associated to low apo AI fractional catabolic Rates
in vivo.
Clin Chim Acta, 2007, 378:53-58.
Barbaux S, Tregouet DA, Nicaud V, Poirier O, Perret C, Godefroy T,
Francomme C, Combadiere C, Arveiler D, Luc G, Ruidavets JB,
Evans AE, Kee F, Morrison C, Tiret L, Brand-Hermann SM, Cambien F.
Polymorphisms in 33 inflammatory genes and risk of myocardial
infarction – A system genetics approach.
J Mol Med 2007, 85:1271-1280.
Quemeneur T, Martin-Nizard F, Kandoussi A, Kyndt X, Vanhille P,
Haculla E, Hatron PY, Fruchart JC, Duriez P, Lambert M.
PON1, a new biomarker of cardiovascular disease, is low in patients with
systemic vasculitis.
Semin Arthritis Rheum, 2007, 37:149-155.
Bouhlel MA, Chinetti-Gbaguidi G, Staels B.
Glitazones in the treatment of cardiovascular risk factors.
Fundam Clin Pharmacol, 2007, 21:7-13.
Tailleux A, Staels B.
Atheroprotective effect of the rexinoid bexarotene in a murine model.
Website, june 2007.
Bouhlel MA, Derudas B, Rigamonti E, Dievart R, Brozek J, Haulon S,
Zawadzki C, Jude B, Torpier G, Marx N, Staels B, Chinetti-Gbaguidi G.
PPARgamma activation primes human monocytes into alternative M2
macrophages with anti-inflammatory properties.
Cell Metab, 2007, 6:137-143.
2008
Brigadeau F, Gele P, Wibaux M, Marquie C, Martin-Nizard F,
Torpier G, Fruchart JC, Staels B, Duriez P, Lacroix D.
The PPARa activator fenofibrate slows down the progression of the left
ventricular dysfunction in porcine tachycardia-induced cardiomyopathy.
J Cardiovasc Pharmacol, 2007, 49:408-415.
Billiet L, Furman C, Cuaz-Perolin C, Paumelle R, Raymondjean M,
Simmet T, Rouis M.
Thioredoxin-1 and its natural inhibitor, vitamin D3 up-regulated protein
1, are differentially regulated by PPARα in human macrophages.
J Mol Biol, 2008, 384 :564-576.
Diabetes, 2007, 56 : 2797-2808.
Billiet L, Furman, Larigauderie G, Copin C, Page S, Fruchart JC,
Brand K, Rouis M.
Enhanced VDUP-1 gene expression by PPARγ agonist induces apoptosis
in human macrophages.
J Cell Physiol, 2008, 214:183-191.
Bouhlel MA, Staels B, Chinetti-Gbaguidi G.
PPAR from active regulators of macrophage biology to pharmacological
targets in the treatment of cardiovascular disease.
J Intern Med, 2008, 263:28-42.
Measuring biomarkers to assess the therapeutic effects of PPAR agonists ?
Pharmacogenomics, 2007, 8:1567-1580.
Bouhlel A, Staels B, Chinetti-Gbagudi G.
Glitazones in the modulation of macrophage lipid homeostasis and the
inflammatory response.
Website, July 2008.
Evans A, Marques-Vidal P, Ducimetiere P, Montaye M, Arveiler D,
Bingham A, Ruidavetz JB, Amouyel P, Haas B, Yarnell J, Ferrieres J,
Fumeron F, Luc G, Kee F, Cambien F.
Patterns of alcohol consumption and cardiovascular risk in Northern
Ireland and France.
Ann Epidemiol, 2007, 17:S75-S80.
Bultel S, Helin L, Clavey V, Chinetti-Gbaguidi G, Rigamonti E,
Colin M, Fruchart JC, Staels B, Lestavel S.
Liver X receptor activation induces the uptake of cholesteryl esters from
high density lipoproteins in primary human macrophages.
Cuaz-Perolin C, Billiet L, Bauge E, Copin C, Scott-Algara D, Genze F,
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Contents
Buchele B, Syrovets T, Simmet T, Rouis M.
Antiinflammatory and antiatherogenic effects of the NF-κB inhibitor
acetyl-11-keto-β-boswellic acid in LPS-challenged apo E-/- mice.
2009
Bougarne N, Paumelle R, Caron S, Hennuyer N, Mansouri R,
Gervois P, Staels B, Haegeman G, De Bosscher K.
Bougarne N, Paumelle R, Haegeman G, Staels B, De Bosscher K.
Circumventing glucocorticoid-mediated hyperinsulinemia via the
activation of PPARα.
Cell Cycle, 2009, 8:2311-2312.
Finck BN, Chinetti G, Staels B.
Editorial : PPARs/RXRs in cardiovascular physiology and disease.
PPAR Res, 2008, 173780.
Bouhlel MA, Brozek J, Derudas B, Zawadzki C, Jude B, Staels B,
Chinetti-Gbaguidi G.
Unlike PPARγ, PPARα or PPARβ/δ activation does not promote human
monocyte differentiation toward alternative macrophages.
Carreon-Torres E, Rendon-Sauer K, Monter-Garrido M, ToledoIbelles P, Gamboa R, Menjivar M, Lopez-Marure R, Luc G, Fievet C,
Cruz D, Vargas-Alarcon G, Perez-Mendez O.
Rosiglitazone modifies HDL structure and increases HDL-apo AI synthesis
and catabolic rate.
Clin Chim Acta, 2009, 401:37-41.
Gineste R, Sirvent A, Paumelle R, Helleboid S, Aquilina A, Darteil R,
Hum DW, Fruchart JC, Staels B.
Phosphorylation of farnesoid X receptor by protein kinase C promotes its
transcriptional activity.
Mol Endocrinol, 2008, 22:2433-2447.
Charton J, Deprez-Poulain R, Hennuyer N, Tailleux A, Staels B,
Deprez B.
Novel non-carboxylic acid retinoids : 1,2,4-oxadiazol-5-one derivatives.
Bioorg Med Chem Lett, 2009, 19/2:489-492.
Legry V, Cottel D, Ferrieres J, Chinetti G, Deroide T, Staels B,
Amouyel P, Meirhaeghe A.
Association between liver X receptor alpha gene polymorphisms and risk
of metabolic syndrome in French populations.
Int J Obes, 2008, 32 :421-428.
Lipid ligand-activated transcription factors regulating lipid storage and
release in human macrophages.
agonists. Part I: synthesis and pharmacological evaluation.
Do HQ, Nazih H, Luc G, Arveiler D, Ferrieres J, Evans A, Amouyel P,
Cambien F, Ducimetiere P, Bard JM.
Influence of cholesteryl ester transfer protein, peroxisome proliferatoractivated receptorα, apolipoprotein E, and apolipoprotein A-I
polymorphisms on high-density lipoprotein cholesterol, apolipoprotein
A-I, lipoprotein A-I, and lipoprotein A-I:A-II concentrations : the
Prospective Epidemiological Study of Myocardial Infarction study.
Metabolism, 2009, 58:283-289.
Cross-talk between statins and PPARalpha in cardiovascular diseases :
clinical evidence and basic mechanisms.
Trends Cardiovasc Med, 2008, 18:73-78.
Dol-Gleizes F, Paumelle R, Visentin V, Mares AM, Desitter P,
Hennuyer N, Gilde A, Staels B, Schaeffer P, Bono F.
Rimonabant, a selective cannabinoid CB1 receptor antagonist, inhibits
atherosclerosis in LDL receptor-deficient mice.
Rigamonti E, Chinetti-Gbaguidi G, Staels B.
Regulation of macrophage functions by PPARalpha, PPARgamma, and
LXRs in mice and men.
Lalloyer F, Pedersen TA, Gross B, Lestavel S, Yous S, Vallez E,.
Rexinoid bexarotene modulates triglyceride but not cholesterol
metabolism via gene-specific permissivity of the RXR/LXR heterodimer in
the liver.
Troughton JA, Woodside JV, Yarnell JWG, Arveiler D, Amouyel P,
Ferrieres J, Ducimetiere P, Patterson CC, Luc G on behalf of the
PRIME Study Group.
Paraoxonase activity and coronary heart disease risk in healthy middleaged males : the PRIME study.
Mysiorek C, Culot M, Dehouck L, Derudas B, Staels B, Bordet R,
Cecchelli R, Fenart L, Berezowski V.
Peroxisome-proliferator-activated receptorα activation protects brain
capillary endothelial cells from oxygen-glucose deprivation-induced
hyperpermeability in the blood-brain barrier.
Curr Neurovasc Res, 2009, 6:181-193.
Zamble A, Martin-Nizard F, Sahpaz S, Hennebelle T, Staels B,
Bordet R, Duriez P, Brunet C, Bailleul F.
Vasoactivity, antioxidant and aphrodisiac properties of Caesalpinia
benthamiana roots.
J. Ethnopharmacol, 2008, 116 :112-119.
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Contents
agonists. Part II: synthesis and pharmacological evaluation of oxime and
acidic head group structural variations.
HDR
Réjane LESTRELIN-PAUMELLE
Rôle et mécanisme d'action moléculaire de PPARalpha dans la réponse
inflammatoire et l'athérosclérose. Rôle et mécanisme d'action
moléculaire de p16INK4a dans le contrôle de la fonction des
monocytes/macrophages dans le syndrome métabolique.
20 juin 2008
Provost AC, Vede L, Bigot K, Keller N, Tailleux A, Jais JP, Savoldelli M,
Ameqrane I, Lacassagne E, Legeais JM, Staels B, Menasche M,
Mallat Z, Behar-Cohen F, Abitbol M.
Morphologic and electroretinographic phenotype of SR-BI knockout mice
after a long-term atherogenic diet.
PhD
Bauge E, Rouille Y, Salles JP, Staels B, Bailleul B.
action in mice.
J Clin Invest, 2009, 119:3830-3838.
Ludivine BILLIET
Directeur de thèse : Mustapha Rouis
« Rôle potentiellement anti-inflammatoire de la thiorédoxine-1 dans
l’athérosclérose : régulation par les récepteurs PPARs au niveau des
macrophages humains »
06 décembre 2007
Tregouet DA, Konig IR, Erdmann J, Munteanu A, Braund PS, Hall AS,
Grosshennig A, Linsel-Nitschke P, Perret C, Desuremain M,
Meitinger T, Wright BJ, Preuss M, Balmforth AJ, Ball SG,
Meisinger C, Germain C, Evans A, Arveiler D, Luc G et al.
Genome-wide haplotype association study identifies the SLC22A3-LPAL2LPA gene cluster as a risk locus for coronary artery disease.
Nat Genet, 2009, 41:283-285.
Mohammed-Amine BOUHLEL
Directeur de thèse : Giulia Chinetti
« Rôle du récepteur nucléaire PPARg dans les macrophages alternatifs
humains : de l’inflammation à l’homéostasie du cholestérol »
10 décembre 2007
Zamble A, Martin-Nizard F, Sahpaz S, Reynaert ML, Staels B,
Bordet R, Duriez P, Gressier B, Bailleul F.
Effects of microdesmis keayana alkaloids on vascular parameters of
erectile dysfunction.
Phytother Res, 2009, 23:892-895.
2010
Lefebvre B, Benomar Y, Guedin A, Langlois A, Hennuyer N,
Dumont J, Bouchaert E, Dacquet C, Penicaud L, Casteilla L,
Pattou F, Ktorza A, Staels B, Lefebvre P.
Proteasomal degradation of Retinoid X Receptorα reprograms
transcriptional activity of PPARγ in obese mice and humans.
J Clin Invest, 2010, 120:1454-1468.
Mayi H, Duhem C, Copin C, Bouhlel MA, Rigamonti E, Pattou F,
Visfatin is induced by peroxisome proliferator-activated receptor gamma
in human macrophages.T.
FEBS J, 2010, 277:3308-3320.
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Contents
• Innate immunity receptors on eosinophils. Human
eosinophils express various receptors, which are able to
recognize molecular structures present on mycobacteria and
tumor cells, in particular TLR2 (Driss et al., 2008) and a
functional γδTCR-CD3 complex (Legrand et al., 2008).
• Hypereosinophilic syndromes (HES). Eosinophils from HES
patients display a decreased oxidative metabolism compared
to the corresponding cells from healthy volunteers.
Immuno-Inflammation
and Cardiovascular
Diseases
David DOMBROWICZ
DR2 Inserm
Immunomodulation
• Role of breast-feeding in prevention of allergic diseases.
Delivery of aerosolized antigen through milk protects
newborns from later developping allergy to the
corresponding antigen (Verhasselt et al., 2008).
• Regulation of allergic diseases by pathogens. Transfer of a B
lymphocyte population isolated from BCG-infected animals
protects recipient mice from the later onset of allergic disease.
• Fractalkine, CX3CL1, and its unique receptor CX3CR1 in Th2associated diseases. CX3CR1 expression by Th2 lymphocytes
controls development of asthma and atopic dermatitis
(Buatois et al., 2008).
• Regulation of asthma and atopic dermatitis by PGD2 and its
receptors. DP1 activation inhibits both asthma and atopic
dermatitis while DP2 activation exacerbates these pathologies
(Angeli et al., 2004; Spik et al., 2005).
• Regulation of asthma atopic dermatitis and anaphylaxis by
PPAR. Both PPARα and γ contribute to the regulation of
asthma (Honda et al., 2004; Woerly et al., 2003) and
anaphylaxis (in this later case, through completely different
mechanisms) (Honda et al., 2008). PPARα and β/δ, but not γ
regulates atopic dermatitis (Staumont-Salle et al., 2008a).
Group Members :
Bouchaert Emmanuel, ITA IPL
Fleury Sébastien, ITA Inserm
Kanda Akira, Postdoctoral fellow
Key words :
Immunology. Metabolism. Genetically modified organism.
Inflammation. Nuclear receptors.
A - Research Report
Physiopathology of allergic reactions :
role of Fc receptors, effector cells and
regulatory mechanisms
During the 2003-2008 period, our projects were focused along
two main lines. On one hand, we have further investigated the
contribution of Fc receptors (FcR), mast cells and eosinophils,
“effectors” in allergic diseases and in anti-tumor immunity. On
the other hand, we have examined various immunotherapeutic strategies in order to prevent allergy
development. Our studies led to the results summarized
below.
Fc receptors
• Role of FcεRI, high affinity IgE receptor, FcγRIII/CD16, low
affinity IgG receptor, and FcεRII/CD23, low affinity IgE receptor
in atopic dermatitis. FcεRI and FcγRIII differentially but partially
regulate pathology onset. FcεRI acts on both Th1 and Th2
responses while FcγRIII only regulates the Th2 response
(Abboud et al., 2008). Absence of CD23 leads to a complete
abrogation of skin inflammation (Staumont-Salle et al., 2008b).
• Role of IgE in anti-tumor immunity. IgE endows monocytes
and eosinophils with anti-tumor cytotoxic activity in vitro.
CD23 expression mediates phagocytosis while FcεRI
contributes to antibody-dependent cellular cytotoxicity
reactions against tumor cells (Karagiannis et al., 2008;
Karagiannis et al., 2007). Low affinity IgE leads to reduced
FcεRI-mediated reactions in vitro and in vivo (Hunt et al., 2008).
development 2010
Mast cells and eosinophils
• Role of the B isoform of inositol-triphosphate kinase (ITPKB)
in allergic reactions. Mast cells from ITPKB-deficient mice are
hyperactivated and barely granulated. This leads to high
histamine concentrations in serum in the absence of overt
inflammation and to a resistance to anaphylaxis.
• IgA receptors on eosinophils. Human, mouse and rat
eosinophils each express a different combination of the 5
molecular structures known to bind IgA (Decot et al., 2005).
Building on our experience in the field of immunoinflammation and allergic diseases and on our longstanding
and successful collaboration with Bart Staels about the
immuno-regulatory role of PPARs in asthma and atopic
dermatitis, we have decided to join this team to develop
research on the role of immuno-inflammation in cardiometabolic diseases, in particular atherosclerosis.
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Indeed, while macrophage contribution to the disease is
clearly established, the role of other immune cell types is not
widely documented. Likewise, studies on expression and
function of nuclear receptors within the immune system
remains only very fragmentary.
Our project will thus investigate the regulation by PPARγ,
expressed by most cells within the immune system, of mast
cells and B lymphocyte contribution to atherosclerosis
physiopathology.
To this aim, we will first study the effects of PPARγ agonists
and antagonists on mast cell and B cell function in vitro. PPARγ
effects on these two cell types will be next investigated in vivo
by specific inactivation of PPARγ in B cells and by adoptive
transfer of PPARγ-deficient mast cells to mast cell-deficient
animals. Consequences of a high-fat diet on the development
of cardio-vascular diseases in these two mouse lines will be
compared to the outcome respectively observed in wild-type
(WT) animals and in mast cell-deficient animals reconstituted
with PPARγ-proficient mast cells. Histological, biochemical and
molecular analyses will be performed.
In order to corroborate the results obtained on animal models,
we will phenotypically, and if possible functionally,
characterize mast cells and B lymphocytes in blood, aortic
lesions and adipose tissue from obese or diabetic patients
treated or not with PPARγ agonists.
In order to determine the potential contribution of nuclear
receptors, besides PPARγ, to the regulation of the immune
system in mice, we will then analyse the expression of
the various nuclear receptor family members in a wide array
of immune cells, in particular B and T lymphocytes, mast cells,
neutrophils and dendritic cells. Based on this transcriptomic
study, we will analyse the contribution of the two nuclear
receptors with the highest expression in B cells and mast cells
using the same experimental strategy as for PPARγ.
Finally, in order to analyse the impact of metabolic alterations,
diet-induced or spontaneously occurring in geneticallyengineered animals generated in the laboratory, on the
immune system, we will also setup an immunophenotyping
platform that will perform extensive histological and flow
cytometry analyses as well as functional tests.
This dual approach should not only allow us to better
delineate the previously underestimated contribution of the
immune system to cardiovascular diseases but also to
evidence metabolism-induced dysregulations of the immune
system.
2008
Hunt J, Bracher MG, Shi J, Fleury S, Dombrowicz D, Gould HJ, Sutton
BJ, Beavil AJ.
Attenuation of IgE affinity for FcepsilonRI radically reduces the allergic
response in vitro and in vivo.
J Biol Chem, 2008, 283 :29882-29887.
Karagiannis SN, Bracher MG, Beavil RL, Beavil AJ, Hunt J, McCloskey
N, Thompson RG, East N, Burke F, Sutton BJ, Dombrowicz D, Balkwill
FR, Gould HJ.
Role of IgE receptors in IgE antibody-dependent cytotoxicity and
phagocytosis of ovarian tumor cells by human monocytic cells.
Cancer Immunol Immunother, 2008, 57:247-263.
Mazuc E, Villoutreix BO, Malbec O, Roumier T, Fleury S, Leonetti JP,
Dombrowicz D, Daeron M, Martineau P, Dariavach P.
A novel drug-like Syk binder prevents anaphylactic shock when
administered orally.
J Allergy Clin Immunol, 2008, 122: 188-194.
Roumier T, Capron M, Dombrowicz D, Faveeuw C.
Pathogen induced regulatory cell populations preventing allergy through
the Th1/Th2 paradigm point of view.
Immunol Res, 2008, 40:1-17.
Staumont-Salle D, Abboud G, Brenuchon C, Kanda A, Roumier T,
Lavogiez C, Fleury S, Remy P, Papin JP, Bertrand-Michel J, Terce F,
Staels B, Delaporte E, Capron M, Dombrowicz D.
Peroxisome proliferator-activated receptor alpha regulates skin
inflammation and humoral response in atopic dermatitis.
Verhasselt V, Milcent V, Cazareth J, Kanda A, Fleury S, Dombrowicz D,
Glaichenhaus N, Julia V.
Breast milk-mediated transfer of an antigen induces tolerance and
protection from allergic asthma.
Nat Med, 2008, 14:170-175.
2009
Abboud G, Staumont-Sallé D, Kanda A, Roumier T, Deruytter N,
Lavogiez C, Fleury S, Rémy P, Papin JP, Capron M, Dombrowicz D.
FcεRI and FcγRIII/CD16 differentially regulate atopic dermatitis in mice.
J Immunol, 2009, 182:6517-6526.
Driss V, Legrand F, Hermann E, Loiseau S, Guerardel Y, Kremer L,
Adam E, Woerly G, Dombrowicz D, Capron M.
TLR2-dependent eosinophil interactions with mycobacteria : role of α-defensins.
Blood, 2009, 113:3235-3244.
Kanda A, Driss V, Hornez N, Abdallah M, Roumier T, Abboud G,
Legrand F, Staumont-Sallé D, Quéant S, Bertout J, Fleury S, Rémy P,
Papin JP, Julia V, Capron M, Dombrowicz D
Eosinophil-derived IFN-γ induces airway hyper responsiveness and lung
inflammation in the absence of lymphocytes.
Publications
2007
Bracher M, Gould HJ, Sutton BJ, Dombrowicz D, Karagiannis SN.
Three-colour flow cytometric method to measure antibody-dependent
tumour cell killing by cytotoxicity and phagocytosis.
J Immunol Methods, 2007, 323:160-171.
Héliot L, Matto V, Soncin F, Gougeon ML, Dombrowicz D, Capron M
A functional γδ TCR/CD3 complex distinct from γδ T cells is expressed by
human eosinophils.
PLoS One, 2009, 4:5926.
Karagiannis SN, Bracher MG, Hunt J, McCloskey N, Beavil RL,
Beavil AJ, Fear DJ, Thompson RG, East N, Burke F, Moore RJ,
Dombrowicz D, Balkwill FR, Gould HJ.
IgE-antibody-dependent immunotherapy of solid tumors : cytotoxic and
phagocytic mechanisms of eradication of ovarian cancer cells.
Nigro EA, Brini AT, Soprana E, Ambrosi A, Dombrowicz D, Siccardi
AG, Vangelista L
Antitumor IgE adjuvanticity : key role of Fc epsilon RI.
J Immunol, 2009, 183:4530-4536.
J Immunol, 2007, 179:2832-2843.
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2010
Dubois A, Deruytter N, Adams B, Kanda A, Delbauve S, Fleury S,
Torres D, François A, Pétein M, Goldman M, Dombrowicz D,
Flamand V.
Regulation of Th2 responses and allergic inflammation through
bystander activation of CD8+ T lymphocytes in early life.
J Immunol, 2010, 185:883-891.
Legrand F, Driss V, Delbeke M, Loiseau S, Hermann E, Dombrowicz D,
Capron M.
Human eosinophils exert TNF-α and granzyme A-mediated tumoricidal
activity towards colon carcinoma cells.
J Immunol, 2010, in press.
Mionnet C, Buatois V, Kanda A, Cossalter G, Milcent V, Fleury S,
Hessel E, Coffman RL, Dombrowicz D, Glaichenhaus N, Julia, V.
CX3CR1 is required for allergic airway inflammation by promoting Th2
cell survival and maintenance in inflamed lung.
Nat Med, 2010, in press.
Mosconi E, Rekima A, Seitz-Polski B, Kanda A, Fleury S, Tissandie E,
Monteiro R, Dombrowicz D, Julia V, Glaichenhaus N, Verhasselt V.
Breast milk immune complexes are potent inducers of oral tolerance in
neonates and prevent asthma development.
Mucosal Immunol, 2010, 3:461-474.
Navarro S, Cossalter G, Chivaroli C, Kanda A, Fleury S, Cazareth J,
Sparwasser T, Dombrowicz D, Glaichenhaus N, Julia V.
The oral administration of bacterial extracts prevents asthma via the
recruitment of regulatory T cells to the airways.
Mucosal Immunol, 2010, in press.
PhD
Georges ABBOUD
Directeur de thèse : David Dombrowicz
« Rôle de FcεRI, CD16 et PPAR-α dans la dermatite atopique »
15 mai 2008
Delphine STAUMONT-SALLE
Directeurs de thèse : Monique Capron and David Dombrowicz
« Mécanismes immunologiques de la dermatite atopique »
10 juin 2008
Fanny LEGRAND
Directeurs de thèse : Monique Capron et David Dombrowicz
« Eosinophiles et immunité innée : expression et rôle de TCR γδ dans la
cytotoxicité anti-tumorale »
21 novembre 2008
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NRs, the peroxisome proliferator-activated receptor alpha
(PPARα) and the retinoic acid receptor alpha to highlight
molecular mechanisms underlying these dual transcriptional
properties, as well as their relative importance in
pathophysiology. Through a mechanism resembling the
canonical model proposed for transrepression, we
demonstrated that both PPARα and the progesterone
receptor can exert anti-proliferative actions by upregulating
cyclin-dependent kinase inhibitors gene expression.
Conferring additional control by endocrine or dietary cues
upon tethering NRs to transcriptional regulatory complexes is
a concept that we extended to nuclear orphan receptors,
which are also important regulators of metabolism and
cellular proliferation. The role of several transcriptional
comodulators of NRs has been explored to unravel novel
potential roles of NRs, as well as to identify mechanisms
restricting the activity of NRs in pathological conditions.
Finally, considering that NRs are established or potentially
valuable pharmacological targets, respectively, in the
treatment of different factors involved in cardiometabolic
diseases, we have generated novel genetic models to
investigate thoroughly their biological properties and to
provide original and relevant tools for drug discovery. These
results are the substratum for our research project.
Molecular Analysis
of Gene Regulation
in Cardiometabolic
Diseases
Philippe Lefebvre
DR2 Inserm
Group Members :
Aumercier Pierrette, MCU Lille 2
Gross Barbara, MCU Lille 2
Helleboid-Chapman Audrey, MCU Lille 2
Dehondt Héléne, ITA Lille 2
Gheeraert Céline, ITA Lille 2
Ploton Maheul, ITA Inserm
Rommens Corinne, ITA IPL
Alexandre Jérémy, ITA Ass. Privée
Oger Frédérik, Postdoctoral fellow
Bensalem Wassym, PhD Student Lille 2
Berrarrah Wahiba, PhD Student Lille 2
Lien Fleur, PhD Student Lille 2
Pawlak Michal, PhD Student Lille 2
Porez Geoffrey, PhD Student Lille 2
Key words :
Molecular biology. Endocrinology. Complexity of the genome.
Molecular and cellular pharmacology. Nuclear receptors.
Transcriptional regulation.
A - Research Report
During the past 4 years, our laboratory has devoted significant
efforts in projects studying the role of, and mechanisms by
which nuclear receptors (NRs) control the transcriptional
activity of genes, and how these NR-mediated responses
impact on metabolic and proliferative cellular responses to
therapeutic and environmental cues. NRs are the core of
various multiproteic complexes bearing enzymatic activities
orchestrating the assembly of the transcriptional machinery
at regulated promoters. The molecular composition of these
complexes is altered in a tissue-specific manner and as a
function of the structure of NR ligands, which are small
lipophilic molecules triggering conformational changes into
NRs. Taking into account the fascinating ability of small ligands
to affect the transcriptional properties of macromolecular
complexes, and, in the long term, to promote adaptative
responses of an organism to dietary or environmental
challenges, we considered NRs as a paradigm to study
intimate mechanisms of transcriptional regulation, as well as
valuable therapeutic targets in the field of metabolic and
proliferative diseases. NRs are two-faceted proteins exhibiting
distinct types of activities, acting either through direct binding
to DNA or by interacting with DNA-bound transcription
factors. Although the transcriptional output can be in each
case either positive or negative, the first type of interaction is
usually thought of as an activating process, whereas the
second is the basis for transcriptional repression. This
dichotomic behaviour has been particularly studied for two
development 2010
We will study the biological and molecular mechanisms
controlling the development of atherosclerosis in order to
develop preventive and therapeutic strategies against its
cardiovascular complications, and study metabolic risk factors,
such as dyslipidemia and type 2 diabetes. We will focus on the
regulation of genes involved in these pathologies and on the
consequences of their dysregulation, studying the molecular
basis and the pathophysiological involvement of nuclear
receptors, transcription factors which our Unit has been
studying for several years. Four different teams will be
organized around a Unit project entitled “Nuclear receptors,
cardiovascular diseases and diabetes”. Team 1 (Nuclear
receptors in the metabolic syndrome) will study the metabolic
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functions of nuclear receptors (FXR, Rev-erbα, RORα) and their
interest as potential therapeutic targets in humans. Team 2
(Molecular control of monocyte/macrophage functions in the
cardiometabolic syndrome) will focus its activities on the role
of PPARg in peripheral blood mononuclear cells from obese
subjects, and will study the role of PPARs, LXRs and the cyclindependent kinase inhibitor CDKN2Ap16INK4a in the molecular
functions in the vascular wall and adipose tissues during
aobesity. The mechanisms by which nuclear receptors control
the contribution of other cells from the immune system to
atherosclerosis will be studied by team 3 (Immunoinflammation and cardiovascular diseases) with a particular
interest for the control by PPARγ of mast cell and B lymphocyte
functions. Team 4 (Molecular analysis of gene regulation in
cardiometabolic diseases) will investigate how nuclear
receptors control gene expression and affect metabolic
functions, allowing the unraveling of novel regulatory
pathways and the validation of nuclear receptors as
pharmacological targets. Our Unit possesses all required
expertise in cellular and molecular biological techniques, and
benefits from comprehensive technical platforms in
biochemistry and histology. The Unit has also a longstanding
experience in the development and study of new animals
models in particular genetically-modified mice.
2008
Carpentier R, Sacchetti P, Segard P, Staels B, Lefebvre P.
The glucocorticoid receptor is a co-regulator of the orphan nuclear
receptor Nurr1.
J Neurochem, 2008, 104:777-789.
Mansouri RM, Bauge E, Gervois P, Fruchart-Najib J, Fievet C,
Staels B, Fruchart JC.
Atheroprotective effect of human apolipoprotein A5 in a mouse model
of mixed dyslipidemia.
Circ Res, 2008, 103:450-453.
Mansouri R, Bauge E, Staels B, Gervois P.
Systemic and distal repercussions of liver-specific PPARa control of the
acute phase response.
Endocrinology, 2008, 149:3215-3223.
Publications
Nowak M, Helleboid-Chapman A, Jakel H, Moitrot E, Rommens C,
Pennachio LA, Fruchar-Najib J, Fruchart JC.
Glucose regulates the expression of the apolipoprotein A5 gene.
J Mol Biol, 2008, 380:789-798.
2007
Ahituv N, Akiyama J, Chapman-Helleboid A, Fruchart J, Pennacchio LA.
In vivo characterization of human apo A5 haplotypes.
Genomics, 2007, 90:674-679
Diabetes, 2007, 56 : 2797-2808.
2009
Bougarne N, Paumelle R, Caron S, Hennuyer N, Mansouri R,
Gervois P, Staels B, Haegeman G, De Bosscher K.
Fontaine C, Rigamonti E, Nohara A, Gervois P, Teissier E, Fruchart JC,
Liver X receptor activation potentiates the lipopolysaccharide
response in human macrophages.
Circ Res, 2007, 101:40-49.
Helleboid-Chapman A, Nowak M, Helleboid S, Moitrot E,
Rommens C, Dehondt H, Heliot L, Drobecq H, Fruchart-Najib J,
Fruchart JC.
Apolipoprotein AV modulates insulin secretion in pancreatic β-cells
through its interaction with Midkine.
Gervois P, Fruchart JC, Staels B.
Drug insight : mechanisms of action and therapeutic applications for
agonists of PPAR.
Nat Clin Pract Endocrinol Metab, 2007, 3:145-156.
Lalloyer F, Pedersen TA, Gross B, Lestavel S, Yous S, Vallez E,
Rexinoid bexarotene modulates triglyceride but not cholesterol
metabolism via gene-specific permissivity of the RXR/LXR heterodimer in
the liver.
Gross B, Staels B.
PPAR agonists : multimodal drugs for the treatment of type-2 diabetes.
Best Pract Res Clin Endoc Met, 2007, 21:687-710.
Lefebvre P, Cariou B, Lien F, Kuipers F, Staels B.
Role of bile acids and bile acid receptors in metabolic regulation. Physiol
Rev, 2009, 89:147-191.
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2010
Carmona MC, Lefebvre P, Lefebvre B, Galinier A, Benani A,
Jeanson Y, Louche K, Flajollet S, Ktorza A, Dacquet C, Pénicaud L,
Casteilla L From The Consortium Of The French Ministry Of Research
And Technology : Molecules And New Therapeutic Targets
Coadministration of coenzyme Q prevents rosiglitazone-induced
adipogenesis in ob/ob mice.
Int J Obes, 2009, 33:204-211.
Lefebvre B, Benomar Y, Guedin A, Langlois A, Hennuyer N,
Dumont J, Bouchaert E, Dacquet C, Penicaud L, Casteilla L,
Pattou F, Ktorza A, Staels B, Lefebvre P.
Proteasomal degradation of Retinoid X Receptorα reprograms
transcriptional activity of PPARγ in obese mice and humans.
J Clin Invest, 2010, 120:1454-1468.
Lefebvre P, Benomar Y, Staels B.
Retinoid X receptors : common heterodimerization partners with distinct
functions.
Trends Endocrinol Metab, 2010, in press.
SUMOylation of the human peroxisome proliferator-activated receptorα
inhibits trans-activity through the recruitment of the nuclear corepressor
NCoR.
J Biol Chem, 2010, 285:5983-5992.
Ristea Popescu I, Helleboid-Chapman A, Lucas A, Vandewalle B,
Dumont J, Bouchaert E, Derudas B, Kerr-Conte J, Caron S, Pattou F,
Staels B.
The nuclear receptor FXR is expressed in pancreatic β-cells and protects
human islets from lipotoxicity.
FEBS Letters, 2010, 584:2845-2851.
PhD
Rodolphe CARPENTIER
Directeur de thèse : Philippe Lefebvre
« Partenaires d’interactions, gènes cibles et rôles potentiels du récepteur
nucléaire orphelin Nurr1 »
26 novembre 2007
Roxane MANSOURI
Directeur de thèse : Philippe Gervois
« PPARα : contribution globale dans le contrôle de l’inflammation liée aux
risques cardiovasculaires »
17 décembre 2007
Benoît POURCET
Directeur de thèse : Corine Glineur
« Mécanismes moléculaires de la régulation de l’activité du récepteur
nucléaire humain PPARα : un rôle-clef des modifications posttraductionnelles »
30 mars 2009
165
Contents
Cross-Sectional
and Emerging
166
Contents
Biostructures
and Molecular
Drug Discovery
Inserm U 761
affiliated to IFR 114 and to IFR 142
Benoît DEPREZ
Group Members :
Déprez Benoît, PU Lille 2
Beghyn Terence, MCU Lille 2
Couturier Cyril, MCU Lille 2
Charton Julie, MCU Lille 2
Déprez-Poulain Rébecca, MCU Lille 2
Flipo Marion, MCU Lille 2
Gesquière Jean-Claude, PU Lille 2
Gras-Masse Hélène, PU Lille 2
Tartar André, PU Lille 2
Willand Nicolas, MCU Lille 2
Leroux Florence, IR Inserm
ElBakali Jamal, ATER Lille 2
Totobenazara Jane, ATER Lille 2
Aknin Karen, Postdoctoral fellow Lille 2
Piveteau Catherine, Postdoctoral fellow Lille 2
Maingot Lucie, Postdoctoral fellow Lille 2
Bourin Arnaud, Postdoctoral fellow Lille 2
Cousaert Nicolas, Postdoctoral fellow Lille 2
Dutot Laurence, Postdoctoral fellow Lille 2
Gluszok Sébastien, Postdoctoral fellow Lille 2
Host Hélène, Postdoctoral fellow Inserm
Jida Mouhamad, Postdoctoral fellow Lille 1
Laconde Guillaume, Postdoctoral fellow Inserm
Pusica Daniça, SAR Lille 2
Vandeputte Tatiana, SAR Lille 2
Dumont Julie, AI Lille 2
Campagne Gabriel, Technician Lille 2
Dassonneville Sandrine, Technician IPL
Deguine Noémie, Technician Lille 2
Landry Valérie, Technician IPL
Matusiak Jennifer, Technician IPL
Gauriot Marion, PhD Student Lille 1
Malaquin Sandra, PhD Student Lille 2
Villemagne Baptiste, PhD Student Lille 2
Desruelle Carole, Assistant IPL
Contact :
00 33 3 20 96 40 24
[email protected]
Key Words :
Chemical genetics. Target validation . Structure-based design.
Chemical library. High throughout screening. Drug discovery.
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Contents
our compound in conditions that mimic an in vivo
environment (plasma, liver microsomes) as well as in whole
animals (PK). ADME data generated all along the project
lifetime are used to optimise the structure of our compounds
and better interpret the results obtained in animal models.
Moreover, we benefit from a close proximity with a team of XRay crystallographers and molecular biologists (lead by
Vincent Villeret, see Figure 2) who help us in the TB project,
providing important structural information. As far as X-ray
crystallography is concerned we also collaborate actively with
the team of Prof. Tang form Chicago (see figure 1).
A - Research Report
U761 “Biostructures & Drug Discovery”
(www.u761.lille. inserm.fr ) was created by Inserm
on January 1st, 2006, with the goal of translating
fundamental biology into drug therapies, in
collaboration with biologists. Our labs (700m2) are
located at the Pasteur Institute and at the Faculty of Pharmacy.
Our researchers are medicinal chemists, analysts and biologists
specialized in screening and ADME. They role, in interdisciplinary
projects, is to design chemical tools to better understand the
function of proteins genetically or functionally related to human
diseases, and to turn, in fine, some of these compounds into drug
candidates that have a new mode of action.
The project portfolio. We manage a portfolio of projects
matching the size of our team and covering different risk
profiles. All our project aims at creating scientific value, but
also medical and business opportunities in well defined time
frames. This is why our projects rely on different sources of
information and techniques and target several disease areas.
In the past 3 years, we have obtained compounds at different
stages in the different projects, from hit to advanced lead. With
this well-balanced portfolio, a broad screening platform
including primary assays and eADME, the odds of delivering
data that will be translated to the clinic and business
environments are maximised.
A visual representation of the portfolio can be found
hereafter :
Figure 2 (structure of an
ethionamide booster in the
EthR receptor)
Relations with industry and valorisation. Pharmaceutical and
biotech companies are prominent actors of drug discovery and
development, and therefore privileged partners for our
activities and projects. Our scientists have been, and still are, in
close contact with industry to remain exposed to industrial
needs and propose innovative solutions to industries and at the
same time learn from them. As an illustration of this policy, from
2003 to 2007, we have collaborated with Fournier Pharma
(Paris) and Euroscreen (Brussels). We currently have two
discovery partnerships with industry (Cytomics System and
Targeon), supported by two ANR-biotech grants. Two of our
internal projects are at a mature state and been granted two
ANR-emergence sponsorships in 2006 and 2007 (“drug boosters
in TB”, “drug morphing in malaria”). The TB project is currently in
a start-up feasibility phase. The new project on non sense
mediated mRNA decay is also very promising: two families of
compounds have been successfully tested in cell and animal
models of Duchenne disease.
Management. Our laboratory has two internal committees.
1/ the Research Operation Committee, is composed of the
permanent researchers (project & platform leaders), the
operation manager and is lead by the director of the lab. The
ROC meets every Monday morning after the Science Meeting.
It examines all questions relative to daily operations and
strategic decisions. In this meeting, every 3 months, the
project and platform leaders are given an update in the
budget (overhead and direct costs).
Key achievements of these projects are summarized in the
following table :
Designing compounds for in vivo proof-of-concept.
Medicinal chemistry is at the heart of our activity. Medicinal
chemists design and synthesize compounds to restore or
modify specifically one or more functions in a whole organism
(animal or human). The chemists need, at the same time, data
on activity, and on physical properties, metabolic stability,
bioavailability (ADMEt).
Thanks to the financial support of Région Nord-Pas-de-Calais,
Institut Pasteur de Lille, U.Lille2 and Inserm as of January 2006,
I have been able to gather a multidisciplinary research team
that can endeavour to design and select bioactive compounds
for the validation both in vitro and in vivo of novel therapeutic
concepts. Our research benefit from a High Throughput
Screening lab, a large library of organic compounds (lead-like
or drug-like), and a significant physical and human resources
dedicated to medicinal chemistry. One of the key assets of our
lab is the ADME platform that allows us to measure the fate of
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2/ a “conseil de laboratoire”, composed of the ROC and elected
members represented all categories of personnel in the lab.
This committee is called at least every year.
Regional and national joint initiatives on drug discovery
and target validation.
PRIM (www.drugdiscoverylille.org) is a consortium dedicated
to the science of drug discovery and lead by Benoit Déprez. It
has been created for a period of 7 years by the Région Nord Pasde-Calais to promote the collaboration between more than 70
biologists, chemists and analysts, and enable target validation
and drug discovery projects in the RŽgion. As a founder
member of PRIM, our team submits new projects to a Scientific
Advisory Board composed of prominent personalities from
academia or recently retired from the pharmaceutical industry
(Sanofi-Aventis, Merck, Fournier, Pfizer).
Our lab is a also founder member of C-DITHEM (www.cdithem.fr)
a national consortium for drug discovery on protein-protein
interaction, together with the labs directed by Fabien Calvo
and Bruno Villoutreix, in Paris and the crystallography team
lead by Vincent Villeret at the Pasteur Institute (Lille).
Quality, Welfare and Safety. We follow working guidelines
that cover quality of data produced, welfare and safety
of scientific, technical and administrative staff. Assay
developments, library synthesis and screening campaigns are
performed according to good laboratory practices. A quality
system was implemented that ensures the quality of the
produced data as well as their traceability. In the end, results
are safely recorded in our password protected database and
a report is issued.
projects (metallohydrolases and protein-protein interactions)
should deliver compounds with a proven efficacy in in vivo
disease models. For Protein protein interaction, a grant
application to ARC has been filed. For IDE, a application has
been filed to ANR, with the teams of Bart Staels and Luc Buée.
Two advanced courses at the frontier of science, medicine
and pharma business. We are very much involved in the
development of new courses and teaching methods. In
particular, with André Tartar, we will continue to develop two
advanced courses in drug discovery and development, given
to Master students and to students in pharmacy (cursus
“industry”). New concepts are taught using drug discovery
and development case studies and personal work. The
objective of the first course is to give a comprehensive
knowledge of the drug discovery methods and techniques.
The second course means to provide a vision and
understanding of the specificity of the pharmaceutical
industry and business, and to give to the students the ability
to use their scientific and medical knowledge for project
evaluation and business development.
These courses are linked to practical work in private
companies in R&D departments, or in academic labs.
With this portfolio of projects, a broad screening platform
including primary assays and eADME, the odds of
delivering data that will be translated to the clinic and
business environments are maximised.
Publications
development 2010
2007
Beghyn T, Hounsou C, Deprez BP.
PDE5 inhibitors : An original access to novel potent arylated analogues
of tadalafil.
Bioorg Med Chem Letters, 2007, 17:789-792.
Research strategy Our research strategy, implemented since
Jan 2006, will be continued. The 2008 portfolio maintained,
hoping that the most advanced projects will be licensed or
spun out and replaced by projects issued from our recent work.
Strengthening our role in translational research. Our
strategy will remain to strengthen our team of expert
scientists in screening, medicinal chemistry and eADME, at the
interface between basic biology and drug discovery. We will
continuously foster collaborations with scientists who master
the biology of novel therapeutic targets to produce drug
candidates (drug discovery) or compounds that help
understand the role of the target (chemical biology). A
portfolio of projects at various stages and risk profiles will be
maintained throughout the 4 next years.
Member of two research federations (IFR). As a sign of the
relevance of medicinal chemistry to the biology community,
in the next period, our team will be a component of 2 research
federations (IFR) in Lille: IFR142 (Molecular and Cellular
Medicine), on the campus of the Pasteur Institute, and IFR 114
(Institute de Medicine Predictive et Therapeutique, on the site
of Regional University Hospital).
Advanced projects. The four projects lauched in the previous
period will be continued and financed. The most advanced
ones (TB, proteasome, NMD) should deliver development
candidates within the next period. For the TB project an grant
application has been filed at the EC (FP7). The more exploratory
Beghyn T, Deprez-Poulain RF, Willand N.
Drug Discovery and Selection - 43rd Rencontres Internationales de Chimie
Thérapeutique, Lille, France.
IDDB MEETING REPORT, 2007.
Charton J, Cousaert N, Bochu C, Willand N, Deprez B, DeprezPoulain R.
A versatile solid-phase synthesis of 3-aryl-1,2,4-oxadiazolones and
analogues.
Tetrahedron Letters, 2007, 48:1479-1483.
Deprez-Poulain RF, Charton J, Leroux V, Deprez BP.
Convenient synthesis of 4H-1,2,4-triazole-3-thiols using di-2pyridylthionocarbonate.
Dhulut S, Bourin A, Lannou MI, Fleury E, Lensen N, Chelain E,
Pancrazi A, Ardisson J, Fahy J.
Cyclic Allyl Carbamates in Stereoselective syn SE' prime Processes :
Synthetic Approach to Sarcodictyins and Eleutherobin.
Eur J Organ Chem, 2007, 2007:5235-5243.
Dubs P, Bourel-Bonnet L, Subra G, Blanpain A, Melnyk O, Pinel AM,
Gras-Masse H, Martinez J.
Parallel synthesis of a lipopeptide library by hydrazone-based chemical
ligation.
J Comb Chem, 2007, 9:973-981.
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Vaccher MP, Charton J, Guelzim A, Caignard DH, Bonte JP, Vaccher C.
Preparative Enantiomeric Separation of Potent AMP-Activated Protein
Kinase Activator by HPLC on Amylose-Based Chiral Stationary Phase.
Determination of Enantiomeric Purity and Assignment of Absolute
Configuration.
J Pharmaceut Biomed Analysis, 2008, 46:920-928.
Flipo M, Beghyn T, Leroux V, Florent I, Deprez BP, Deprez-Poulain RF.
Novel Selective Inhibitors of the Zinc Plasmodial Aminopeptidase PfA-M1
as Potential Antimalarial Agents.
J Med Chem, 2007, 50:1322-1334.
Flipo M, Beghyn T, Charton J, Leroux VA, Deprez BP, DeprezPoulain RF.
A library of novel hydroxamic acids targeting the metallo-protease family :
design, parallel synthesis and screening.
Bioorg Med Chem, 2007, 15:63-76.
Villoutreix BO, Bastard K, Sperandio O, Fahraeus R, Poyet JL,
Calvo F, Deprez B, Miteva MA.
In silico-in vitro screening of protein-protein interactions: towards the
next generation of therapeutics.
Curr Pharmaceut Biotechnol, 2008, 9:103-122.
Gras-Masse H, Willand N.
Neuraminidase inhibitors and risk of H5N1 influenza.
Annales Pharmaceutiques Françaises, 2007, 65:50-57.
2009
Gilleron P, Wlodarczyk N, Houssin R, Farce A, Laconde G,
Goossens JF, Lemoine A, Pommery N, Henichart JP, Millet R.
Design, synthesis and biological evaluation of substituted
dioxodibenzothiazepines and dibenzocycloheptanes as farnesyltransferase inhibitors.
Charton J, Leroux F, Delaroche S, et al.
Synthesis of a 200-member library of squaric acid N-hydroxylamide
amides (vol 18, pg 4968, 2008).
Charton J, Deprez-Poulain R, Hennuyer N, Tailleux A, Staels B,
Deprez B.
Novel non-carboxylic acid retinoids : 1,2,4-Oxadiazol-5-one derivatives.
Host H, Drobecq H, Locht C, Menozzi FD.
FEMS Microbiology Letters, 2007, 268:144-150.
Flipo M, Charton J, Hocine A, Dassonneville S, Deprez B, DeprezPoulain R.
Hydroxamates : Relationships between structure and plasma-stability.
J Med Chem, 2009, 52:6790-6802.
Telliez A, Desroses M, Pommery N, Briand O, Farce A, Laconde G,
Lemoine A, Depreux P, Henichart JP.
Derivatives of Iressa, a specific epidermal growth factor receptor inhibitor,
are powerful apoptosis inducers in PC3 prostatic cancer cells.
Chem Med Chem, 2007, 2:318-332.
Klose D, Laprais M, Leroux V, Siepmann F, Deprez B, Bordet R,
Siepmann J.
Fenofibrate-loaded PLGA microparticles : Effects on ischemic stroke.
Eur J Pharmaceut Sci, 2009, 1:43-52.
Willand N, Folléas B, Boutillon C, Verbraeken L, Gesquiere JC, Tartar
A, Deprez, B.
Efficient, Two-Step Synthesis Of N-Substituted Nortropinone Derivatives.
Leroux F, Déprez-Poulain R, Frénois F, Aumercier M, Locht C,
Villeret V, Déprez B, Baulard A.
Synthetic EthR inhibitors boost anti-tuberculous activity of ethionamide.
Nature Med, 2009, 15:537-544.
2008
Beghyn T, Deprez-Poulain R, Willand N, Foleas B, Deprez B.
Natural compounds : leads or ideas ? Bioinspired molecules for drug
discovery.
Chem Biol Drug Design, 2008, 72:3-15.
2010
Jida M, Deprez-Poulain R, Malaquin S, Roussel P, AgbossouNiedercorn F, Deprez B, Laconde G.
Solvent-free microwave-assisted Meyers’ lactamization.
Green Chem, 2010, 12:961-964.
Charton J, Charruault L, Deprez-Poulain R, Deprez B.
Alkylsquarates as key intermediates for the rapid preparation of original
drug-inspired compounds.
Combinat Chem High Throughput Screening, 2008, 11:294-303.
Jida M, Malaquin S, Deprez-Poulain R, Laconde G, Deprez B.
Synthesis of five- and six-membered lactams via solvent-free microwave
Ugi reaction.
Charton J, Leroux F, Delaroche S, Landry V, Deprez BP, DeprezPoulain RF.
Synthesis of a Library of 200-Member Squaric Acid N-Hydroxylamide Amides.
Lipka E, Folly-Klana M, Charton J, Vaccher MP, Bonte JP, Vaccher C.
Determination of pKa values of benzimidazole derivatives from mobility
obtained by capillary electrophoresis.
J Pharmaceut Biomed Analysis, 2010, 53:1267-1271.
Cousaert N, Willand N, Gesquiere JC, Tartar A, Deprez B, DeprezPoulain R.
Original loading and Suzuki conditions for the solid-phase synthesis of
biphenyltetrazoles. Application to the first solid-phase synthesis of
irbesartan.
Maingot L, Leroux F, Landry V, Dumont J, Nagase H, Villoutreix B,
Sperandio O, Deprez-Poulain R, Deprez B.
New non-hydroxamic ADAMTS-5 inhibitors based on the 1,2,4-triazole3-thiol scaffold.
Bioorg Med Chem Letters, 2010, In Press, Accepted Manuscript : In
Press.
Magnier-Bouvier C, Reboule I, Gil R, Collin J.
Samarium Diiodide as an Efficient Catalyst for the Conversion of NAcyloxazolidinones into Esters.
Synlett, 2008, 1211-1215.
Malaquin S, Jida M, Gesquiere JC, Deprez-Poulain R, Deprez B,
Laconde G.
Ugi reaction for the synthesis of 4-aminopiperidine-4-carboxylic acid
derivatives. Application to the synthesis of carfentanil and remifentanil.
Reboule I, Gil R, Collin J.
Enantioselective Conjugate Addition of Aromatic Amines to NAlkenoyloxazolidinones Catalyzed by Iodido-(binaphtholato)samarium.
Eur J Organ Chem, 2008, 532-539.
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Reynes C, Host H, Camproux AC, Laconde G, Leroux F, Mazars A,
Deprez B, Fahraeus R, Villoutreix BO, Sperandio O.
Designing Focused Chemical Libraries Enriched in Protein-Protein
Interaction Inhibitors using Machine-Learning Methods.
Plos Comput Biol, 2010, 6:e1000695.
Willand N, Desroses M, Toto P, Dirie B, Lens Z, Villeret V, Rucktooa P,
Locht C, Baulard A, Deprez B.
Exploring Drug Target Flexibility Using in Situ Click Chemistry : Application
to a Mycobacterial Transcriptional Regulator.
ACS Chem Biol, 2010, In press.
PhD
Nicolas COUSAERT
Directeur de thèse : Rebecca Deprez-Poulain
Conception et synthèse d'inhibiteurs de métalloprotéases et de cibles à
ligand acide
19 novembre 2008
Patents
Deprez B, Willand N, Dirié B, Toto P, Villeret V, Locht C, Baulard AR,
Compounds having a potentiating effet on the activity of ethionamide
and uses thereof,
WO2008003861, 10th Jan. 2008
Deprez B, Beghyn T, Laconde G, Charton J,
Chiral Tetra-hydro beta-carboline derivatives, applications thereof as
antiparasitic compounds,
WO2008044144, 23rd Apr. 2008
Carniato D, Jaillardo K, Busnel O, Gutmann M, Briand J, Deprez B,
Thomas D, Bougeret C,
Composés utiles pour le traitement des cancers,
FR 08 53944, 2008
Deprez-Poulain R, Charton J, Deprez B, Leroux F, Gauriot M, Tang W-J,
Totobenazara J.
Ligands of Insulin Degrading Enzyme and Their Uses,
EP_20100330093425, 4th mar 2010
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Inhibition
of Nonsense-Mediated
mRNA Decay
Groupe Avenir Inserm
Fabrice LEJEUNE
Contact :
00 33 3 20 87 71 21
[email protected]
Group Members :
Lejeune Fabrice, CR Inserm
Fic Weronika, Postdoctoral fellow Inserm
Gonzalez Sara, PhD Student Inserm
Key Words :
RNA. Nonsense-mediated mRNA decay (NMD).
Premature stop codon (PTC). Screening. mRNA stability.
Mammals. Duchenne muscular dystrophy.
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Contents
A - Research Report
P-bodies in HeLa cells
(Dcpla staining)
In order to get the right gene expressions, cells developed
several layers of quality controls check points. One of these
quality controls occurs after mRNA maturation and before the
bulk of translation, and is called nonsense-mediated mRNA
decay (NMD). The roles of NMD are to identify and degrade
mRNAs harboring a premature termination codon (PTC) in
order to protect the cells from deleterious functions from
truncated proteins or to prevent the synthesis of non functional
truncated proteins. Another role of NMD is also to regulate
some gene expression by recognizing a normal stop codon as
a PTC which leads to inhibit the expression of this specific gene.
A stop codon is recognized as a PTC if it localizes at more than
50-55 nucleotides upstream of a splicing event in mammals.
One third of inherited genetic diseases and many cancers
involve apparition of nonsense mutations. In most of cases, the
disease is due to the absence of expression of the mutated gene
because of the activation of NMD on the PTC-containing mRNA.
Sometimes the truncated protein could be as functional as the
full length protein depending on the position of the PTC. It is
for this reason that we are interested by blocking NMD in the
context of human genetic diseases and study the consequences
of the synthesis of truncated proteins. Blocking NMD is also a
very powerful approach to study the regulation of NMD and the
mechanism of NMD in general. Our team started on January
2008 as an INSERM AVENIR group.
Publications
Durand S, Cougot N, Mahuteau-Betzer F, Nguyens CH, Grierson D,
Bertrand E, Tazi J, Lejeune F.
Inhibition of nonsense-mediated mRNA decay (NMD) by a new chemical
molecule reveals the dynamic of NMD factors in P-bodies.
J Cell Biol, 2007, 178:1145-1160.
Dreumont N, Bourgeois CF, Lejeune F, Liu Y, Ehrmann IE, Elliott DJ,
Stévenin J.
Human RBMY regulates germline-specific splicing events by modulating
the function of the serine/arginine-rich proteins 9G8 and Tra2-{beta}.
J Cell Sci, 2010, 123:40-50.
HDR
Lejeune Fabrice
Tuteur : Lemoine Yves
Régulation de la stabilité des ARNm soumis au nonsense-mediated mRNA
decay (NMD).
16 mai 2008
B - Perpectives and
Development 2010
In order to inhibit NMD, we decided to identify small chemical
molecules capable to interfere with the UPF proteins functions
which are essential NMD factors. For that we built a cell
screening system that allows the test of thousand molecules
in a fast way. We also developed tools for the characterization
of NMD inhibitors and to understand the mode of action of
each selected molecules. Finally, we have several patient cell
lines and 2 mouse models to study the effects of NMD
inhibitors in vivo and in the context of genetic diseases. This
work involves collaborations with chemist groups, pharmacologist groups and other molecular and cellular biologist
teams. In 2010 we plan to screen the 30 000 compounds library
from Pr. Benoit Deprez lab and initiate the characterization of
the first hits.
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Microbiological
Safety Unit
Michèle VIALETTE
Contact :
00 33 3 20 87 78 53
[email protected]
Group Members :
Vialette Michèle, Manager IPL
Pinon Anthony, Assistant Manager IPL (Statistics)
Alexandre Virginie, Technician IPL
Caudrelier Yvette, CE IPL
Deboosère Nathalie, CE IPL (Virology)
Decherf Sandra, Technician IPL
Delobel Alexandre, Technician IPL
Gachet Jessica, CE IPL
Key Words :
Microbiology. Bacteriology. Virology. Biothreat agents.
Detection of pathogenic micro-organisms.
Behaviour of pathogenic micro-organisms in the environment.
Food. Water. Air. Mathematical modelling. Molecular biology.
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Contents
The Microbiological Safety Unit specializes in detection, study,
and management of microbial hazards in various
environments: food, water, air, cosmetics, etc., linked to either
accidental or intentional contamination. Work focuses on
Class 2 and 3 pathogenic bacteria and viruses, using
microbiological, molecular, and mathematical tools.
Applied projects include design, optimisation, or validation of
new methods for detection and quantification of pathogenic
micro-organisms in the environment. The unit also performs
studies aiming at evaluating and modelling the behaviour
(growth, persistence, death…) of these pathogens as
influenced by environmental or human factors.
an artificially-introduced bacterial population in the food
product). Such experiments require reliable methods to
inoculate and enumerate the micro-organisms. We
participated in a large study to optimise these methods, and
to obtain quantitative evaluations of the variability of the
microbial behaviour in foods.
Contamination of food products may occur through contact
with working surfaces in production facilities. Evaluation of
the efficiency of cleaning and decontamination procedures
against pathogenic enteric viruses would be facilitated
thanks to the use of surrogate, non-pathogenic viruses. We
have been involved in a study trying to identify the best
models for human pathogenic viruses with respect to the
adhesion on inert or organic surfaces, in order to assess
decontamination procedures of surfaces.
In addition, the Microbiological Safety Unit has been
requested to evaluate detection systems targeting biothreat
agents in the environment, in the framework of confidential
programs.
A - Research Report
Detection of pathogenic microorganisms in the environment
Development 2010
Avian Influenza Viruses, including the Highly Pathogenic
H5N1 subtype, are carried by wild birds and are then found
in their faeces. Hence, surface water or mud from lakes or
ponds are natural reservoirs for the viruses. In the framework
of the European Project RIVERS, with partnership of several
Pasteur Institutes around the world, we optimised methods
for detection of such viruses in large volumes of surface
water and in mud. This method was successfully applied on
samples linked to recent outbreaks in Europe and Asia.
Human infection by Legionella occurs by inhalation of an
aerosol produced from contaminated water and dissipated
in the air. European regulations or guidelines for installations
at risk concern maximal allowable concentrations in water.
However, it seems difficult to firmly evaluate Legionella
presence in aerosols based on water concentrations, in spite
of probable correlations between them. Air monitoring
would therefore appear as the most relevant surveillance
method to evaluate the risk of Legionella contamination. We
optimised a method to collect and analyse air samples. This
operational protocol could then be applied to obtain reliable
evaluations of survival of aerosolised Legionella from
contaminated water.
The Microbiological Safety Unit is a partner in a collaborative
project aiming at the design of an innovative method for the
detection of Legionella pneumophila in water. This method
will be based on a specific enzymatic reaction. It should
provide rapid, on-site detection of Legionella pneumophila in
water networks.
In the field of food safety, we will conduct a study to apply
molecular tools to simultaneously detect and assess the
pathogenicity of Listeria monocytogenes. This foodborne
pathogen represents a serious threat in ready-to-eat food
products, and it is therefore important to evaluate the risk
linked to its presence.
New perspectives include the evaluation of the efficiency of
air filtration systems for removal of airborne microbial
contamination. Clean air represents a crucial issue for public
health, especially in healthcare facilities where patients are
most susceptible to microbial infections.
Regarding bioterrorism agents, European-level projects are
foreseen for the upcoming months.
Influence of environmental factors
on the behaviour of pathogenic
micro-organisms
Publications
An increasing number of toxi-infections are linked to the
consumption of food contaminated by enteric viruses.
Imported frozen berries may be contaminated by Hepatitis
A virus, and thermal treatment is thus used to eliminate this
hazard. To optimise these treatments, influence of pH on the
survival of Hepatitis A virus in ground berries was evaluated.
A predictive model was built to predict inactivation kinetics
of the virus as influenced by temperature and pH.
Assessment of the microbiological stability of food products
throughout their shelf-life uses several tools, including
predictive microbiology and challenge tests (monitoring of
2007
Certad G, Ngouanesavanh T, Guyot K, Gantois N, Chassat T,
Mouray A, Fleurisse L, Pinon A, Cailliez JC, Dei-Cas E, Creusy C.
Cryptosporidium parvum, a potential cause of colic adenocarcinoma.
Infect Agents Cancer, 2007, (22):1-11.
Pinon A, Alexandre V, Cupferman S, Crozier A, Vialette M.
Growth, survival and inactivation of Pseudomonas aeruginosa and
Staphylococcus aureus strains of various origin in presence of Ethanol.
Int J Cosmetic Sci, 2007, 29:111-119.
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Contents
Tresse O, Shannon K, Pinon A, Malle P, Vialette M, MideletBourdin G.
Variable Adhesion of Listeria monocytogenes Isolates from FoodProcessing Facilities and Clinical Cases to Inert Surfaces.
J Food Protection, 2007, 70:1569-1578.
2009
Jiménez JC, Pinon A, Dive D, Capron M, Dei-Cas E, Convit J.
Antibody response in children infected with Giardia intestinalis before
and after treatment with Secnidazole.
Am J Tropical Med Hygiene, 2009, 80:11-15.
2010
Augustin JC, Bergis H, Bourdin G, Cornu M, Couvert O, Denis C,
Huchet V, Lemonnier S, Pinon A, Vialette M, Zuliani V, Stahl V.
Design of challenge testing experiments to assess the variability of Listeria
monocytogenes growth in foods.
Food Microbiol, 2010, doi: 10.1016/j.fm.2010.05.028.
Certad G, Ngouanesavanh T, Guyot K, Gantois N, Chassat T , Mouray
A, Flament N, Fleurisse L, Pinon A, Delhaes L, Dei-Cas E, Creusy C.
Development of Cryptosporidium parvum induced gastro-intestinal
neoplasia in SCID mice: severity of lesions is correlated with infection
intensity.
Am J Tropical Med Hyg, 2010, 82:257-265.
Deboosère N, Pinon A, Delobel A, Temmam S, Morin T, Merle G,
Blaise-Boisseau S, Perelle S, Vialette M.
A predictive microbiology approach for thermal inactivation of Hepatitis
A virus in acidified berries.
Food Microbiol, 2010, 27:962-967.
Lénès D, Deboosère N, Ménard-Szczebara F, Jossent J, Alexandre V,
Machinal C, Vialette M.
Assessment of the removal and inactivation of influenza viruses H5N1
and H1N1 by drinking water treatment.
Water Res, 2010, 44:2473-2486.
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Biology and Diversity
of Emerging Eukaryotic
Pathogens (BDEEP)
EA 4547
Eric VISCOGLIOSI
Contact :
00 33 3 20 87 79 61
Fax : 00 33 3 20 87 78 88
[email protected]
Group Members :
Viscogliosi Eric, CR CNRS
Aliouat El Mouktar, PR Univ Lille 2
Aliouat-Denis Cécile-Marie, MCU Univ Lille 2
Certad Gabriela, MCU Univ Lille 2
Chabé Magali, MCU Univ Lille 2
Dei-Cas Eduardo, MCU-PH Univ Lille 2/CHRU
Delhaes Laurence, MCU-PH Univ Lille 2/CHRU
Demanche Christine, MCU Univ Lille 2
Standaert-Vitse A, MCU Univ Lille 2
Fréalle Emilie, PH Univ Lille 2/CHRU
Guyot Karine, IE IPL
Gantois Nausicaa, Technician IPL
Pottier Muriel, Technician Univ Lille 2
Sanciu Giovanna, Postdoctoral fellow ANR
Benamrouz Sadia, PhD student ICL
Mantini Cléa, PhD student MENRT
Martinez Anna, PhD student MENRT
Méloni Dionigia, PhD student Grant Region of Sardinia
Key Words :
Antioxidant enzymes. Aspergillus. Life cycle, Blastocystis.
Cryptosporidium. Environmental Parasitology and Mycology.
Evolution. Genomic and transcriptomic. Host-parasite
relationships. Molecular epidemiology. Molecular phylogeny
and taxonomy. Nosocomial infection. Parasites and cancer.
Parasitic or fungal environmental risk. Pathogenesis.
Pathophysiology. Pneumocystis. Scedosporium.
Superoxyde dismutases. Taxonomy. Trichomonas.
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Contents
Phylogenetic approaches allowed also to explore the
emergence of pathogenic power in the evolution of the
targeted pathogens, and to assess zoonotic potential, hostpathogen co-evolution and pathogenic power of microbial
genotypes (Pneumocystis, Aspergillus, Scedosporium,
Blastocystis, Cryptosporidium).
A - Research Report
1) Scientific background, collaborations, and research supports
BDEEP research activities are focused on opportunistic
eukaryotic pathogens responsible for emergent or reemergent diseases with growing impact for the last years,
constituting today major public health issues. These agents
are responsible for severe nosocomial diseases, like fungi
Pneumocystis and Aspergillus, or frequent community
infections, like protists Trichomonas, Cryptosporidium or
Blastocystis. Although these pathogens are among the most
frequently encountered by humans worldwide, the diseases
they cause are usually neglected or considered as "orphan" or
"rare" by health authorities, research organizations and the
pharmaceutical industry. Basic knowledge of these
microorganisms is required to understand their circulation in
ecosystems, mechanisms of proliferation and infection,
pathophysiology, to clarify their real biodiversity, and to define
cost-effective, field-adapted rational prevention and control
strategies. Especially, fine analyses of genetic divergence are
needed to track these pathogens in host populations in order
to clarify their transmission patterns. For this reason, our
research project is focused on the basic biology, pathogenic
power and genetic diversity of these parasites. Moreover, few
or no efficient drugs are available against these pathogens.
Therefore, the BDEEP laboratory contributes also to the
development of antimicrobial drugs mainly through the
identification of new molecular targets as few or no efficient
medicaments, which often cause unwished effects, are
available against these pathogens, and resistances are on the
rise. This project strongly associates basic with clinical
research.
Collaborations are developed with numerous French
laboratories and hospitals and groups established in Europe,
Asia, North and South America, and Australia. Research
projects are supported by University of Lille 2, Institute Pasteur
of Lille, French Ministry of Health (three National Clinical
Research Hospital Programs funded), National Research
Agencies (four programs funded by ANR), Programs ECOSNord and ECOS-Sud, Genopoles Ile-de-France and Lille,
Research Grants from Pfizer, “Vaincre la mucoviscidose” and
Japan Society for the Promotion of Science (JSPS), and by
direct support of Society of Medical Parasitology Professors
(ANOFEL network).
Pneumocystis : genetic diversity and life cycle – The genus
Pneumocystis comprises numerous fungal pathogens
dwelling in the lungs of a wide spectrum of mammals. The
airborne transmitted Pneumocystis species are highly hostspecies specific, and a striking matching of Pneumocystis and
host phylogenetic trees has been reported. Therefore, genetic
diversity of Pneumocystis can be used as a taxonomic marker
in order to clarify the as yet uncertain phylogenetic
relationships in some mammal groups such as bats or rodents.
We aim at identifying Pneumocystis organisms in wild rodents
and bats from different areas worldwide especially from South
East Asia and New World Areas. In parallel, we try to determine
Pneumocystis jirovecii reservoir, infection sources and
mechanisms involved in the human infection. Firstly, efficient
PCR assays allowing Pneumocystis detection in invasive or
noninvasive clinical samples were developed and applied
successfully to both Pneumocystis pneumonia (PcP) diagnosis
and Pneumocystis detection in carriers. Secondly, basic
Pneumocystis biology approaches associated with
experimental and clinical research have increased our
understanding of host-to-host Pneumocystis airborne
transmission. Thirdly, though the Pneumocystis pathogenic
power appears clearly linked to the proliferation capacity of
these fungi, data about Pneumocystis multiplication
mechanisms are scarce or controversial. They are being
explored in a transcriptomic/proteomic perspective to clarify
both Pneumocystis life cycle and cell cycle. Finally, a thematic
program around the emergence of P. jirovecii – Trichomonas
co-infection human cases was developed. The report of high
incidence of this pulmonary co-infection is of major interest
in public health because it could be correlated with the global
burden of lung diseases.
2) Principal results
BDPEE research activity covered two main domains: (A): Basic
biology and pathogenic power, specifically, the team has tried
to clarify the Pneumocystis life cycle, focused on the major role
of superoxide dismutases (SODs) enzymes (potential drug
targets) in parasitic protists, and explored host–parasite
relationships in several pathogens; (B) Genetic diversity, as
molecular phylogeny or taxonomy developments led us to
both identify new species and define new taxonomic patterns.
Cryptosporidiosis molecular epidemiology - Important
cryptosporidiosis outbreaks occurred in numerous countries.
Molecular strategies warranted by our group allow to detect
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Contents
low parasite rates and to identify species or genotypes in
animal, human or environmental samples. Two main species
parasitize humans: C. hominis closely adapted to these hosts,
and C. parvum, well adapted to livestock but able to infect
humans or other mammals. Other genotypes in C. parvum,
and other species were revealed to be able to infect humans,
as illustrated by our researches in developed and developing
regions using multilocus approaches. Our team is involved in
the Cryptosporidium ANOFEL network, and has taken in
charge the organization of the Cryptosporidium National
Collection.
Trypanosoma brucei, Plasmodium falciparum, and
Trichomonas vaginalis. Strikingly, parasitic protists colonizing
humans only possess dimeric FeSODs that differ from the
tetrameric MnSOD and Cu/ZnSOD of mammals. A fine analysis
of SOD structures shows differences in residues and nature of
the interactions between residues involved in the monomermonomer interface of these two types of enzymes. Therefore,
it seems possible to disturb the monomer-monomer interface
of dimeric FeSODs of parasitic protists and to prevent active
dimer formation without changing that of human tetrameric
MnSOD.
Cryptosporidium parvum induces digestive neoplasia in
SCID mice – In order to set in our lab a routine passage of
Cryptosporidium species we used dexamethasone-treated
SCID mice orally infected with C. parvum oocysts. The
infection was markedly enhanced by dexamethasone and
mice did shed oocysts until euthanasia. The most impressive
result was the developing of C. parvum induced low or high
grade neoplasia in the digestive tract of either
dexamethasone-treated or untreated SCID mice. All
dexamethasone-treated SCID mice euthanatized after 46 days
post-infection developed caecal polyps with areas of intraepithelial neoplasia. Moreover, some untreated SCID mice
developed also gastric or duodenal low or high grade
neoplastic changes. The severity of the neoplastic process was
correlated with C. parvum growth. Is C. parvum able to induce
neoplastic lesions in humans or other mammals? We have no
data, but a recent paper reported unexpected high
Cryptosporidium prevalence in European patients with colorectal cancer.
Genetic diversity of Blastocystis isolates - We have analysed
the genetic diversity of Blastocystis isolates from different
hosts. Our phylogenetic analyses revealed a considerable
genetic diversity that could be correlated with the existence
of potentially 12 different species within the genus. We also
confirmed the strong zoonotic potential of this parasite and
suggested the existence of a large potential reservoir in
animals for infections in humans. More recently, we have
proposed a standardization of Blastocystis taxonomy and
suggested that all mammalian and avian isolates should be
designated Blastocystis sp. and assigned to one of nine
subtypes. In parallel, we have demonstrated the pathogenicity
of numerous Blastocystis strains and performed studies of the
genetic diversity of French and Egyptian isolates.
Molecular epidemiology of aspergillosis and scedosporiosis - Filamentous fungi, especially Aspergillus
fumigatus and Scedosporium spp, are able to induce lung
colonization, respiratory allergy and/or severe life-threaten
infections. Development of a given clinical form depends on
both host defences and intrinsic fungal properties. The
present project explored this aspect in a double perspective:
a basic biological research approach and a more clinical
analysis. On the basic biological research side, we examine
fungal molecular epidemiology and phylogeny, in order to
analyze the emergence of pathogenic properties throughout
the evolution. These properties are explored by targeting
fungal SODs, which might play a crucial role in the infectious
lung process. In parallel, we develop a transcriptomic
approach to analyze the fungal anti-oxidative stress response
in the framework of the European Aspergillus Transcriptome
Consortium. On the clinical research side, our group is
exploring the role of fungal colonization in cystic fibrosis (CF)
patient evolution based on the first national prospective study
on this topic that we currently conduce. We also are analyzing
the Aspergillus and Scedosporium species diversity according
to clinical and therapeutic data (“Scedosporium ISHAM
international Network), and “fungal risk in cystic fibrosis
(ISHAM international Network).
development 2010
BDEEP is now developing its main research activities on the
parasites Pneumocystis-micromycetes and Blastocystis.
Although taxonomically unrelated (like fungi and protists),
they share common features, especially wide genetic diversity
with significant impact on speciation, pathogenic power or
transmission patterns. Therefore, in a perspective of
integrative biology, the specificity of our project is the
Superoxide dismutases in parasitic protists and
therapeutic targeting – SODs are usually the sole enzymes
able to scavenge toxic superoxide anions resulting from either
cellular metabolism or immune response cell effectors. We
have characterized these enzymes at the molecular and
structural levels in different pathogenic protists such as
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Contents
Fréalle E, Rodrigue M, Gantois N, Aliouat CM, Delaporte E,
Camus D, Dei-Cas E, Kauffmann-Lacroix C, Guillot J, Delhaes L.
Phylogenetic analysis of Trichophyton mentagrophytes human and
animal isolatesbased on MnSOD and ITS sequence comparison.
Microbiology, 2007, 153:3466-3477.
constant aiming at associating to the understanding of the
mechanisms of infection and pathophysiology of diseases
these agents cause, a detailed exploration of the biology and
genetic diversity of these microorganisms. Several biological
questions common to these two groups are raised regarding
i) the prevalence, molecular diversity, and circulation of these
parasites in human and animal populations (reservoirs,
infection sources, zoonotic potential), ii) the characterization
of this genetic diversity (comparative genomic) and its impact
on the pathogenic power and transmission of the identified
species and variants, iii) the molecules and mechanisms
involved in pathogenesis and proliferation of these pathogens
and iv) their interactions with host cells.
Hadrich I, Vandewalle P, Cheickrouhou F, Makni F, Krichen MS,
Vitse A, Ayadi A, Poulain D.
Ethnic and socio-cultural specificities in Tunisia have no impact on the
prevalence of anti-Saccharomyces cerevisiae antibodies in Crohn's
disease patients, their relatives or associated clinical factors.
Scand J Gastroenterol, 2007; 42:717-725.
Harry M, Roy V, Mercier A, Livet A, Garnier E, Bousserrhine N,
Demanche C.
Isolation and characterization of microsatellite markers in Labiotermes
labralis (Isoptera, Termitidae, Nasutitermitinae).
Mol Ecol Notes, 2007, 7:121-123.
After evaluation by the scientific adivisory board of the Center
for Infection and Immunity of Lille (CIIL), the team has
obtained the label «emerging team» of the CIIL since the
second half of 2010.
Jimenez JC, Morelle W, Michalsky JC, Dei-Cas E.
Excreted/secreted glycoproteins of G. intestinalis play an essential role in
the antibody response.
Parasitol Res, 2007, 100:715-720.
Publications
Khayath N, Vicogne J, Ahier A, Ben Younes A, Konrad C, Trolet J,
Viscogliosi E, Brehm K, Dissous C.
FEBS J, 2007, 274:59-676.
2007
Certad G, Ngouanesavanh TM, Guyot K, Gantois N, Chassat T,
Mouray A, Fleurisse L, Pinon A, Cailliez JC, Dei-Cas E, Creusy C.
Cryptosporidium parvum, a potential cause of colic adenocarcinoma.
Infect Agent Cancer, 2007, 2:22.
Lefèvre E, Bardot C, Noël C, Carrias JF, Viscogliosi E, Amblard C,
Sime-Ngando T.
Unveiling fungal zooflagellates as members of freshwater picoeukaryotes :
evidence from a molecular diversity study in a deep meromictic lake.
Environ Microbiol, 2007, 9:61-71.
Chabé M, Sanchez CA, Aliouat EM, Durand-Joly I, López I,
Dei-Cas E, Vargas SL.
Exploring transplacental transmission of Pneumocystis oryctolagi in firsttime pregnant and multiparous rabbit does.
Med Mycol, 2007, 45:701-707.
Meamar AR, Guyot K, Certad G, Dei-Cas E, Mohraz M, Mohebali M,
Mohammad K, Mehbod AA, Rezaie S, Rezaian M.
Molecular characterization of Cryptosporidium isolates from humans
and animals in Iran.
Appl Environ Microbiol, 2007, 73:1033-1035.
Duboucher C, Caby S, Chabé M, Gantois N, Delgado-Viscogliosi P,
Pierce R, Capron M, Dei-Cas E, Viscogliosi E.
Trichomonoses pulmonaires humaines.
Presse Med, 2007, 36:835-839.
Noël C, Noda S, Mantini C, Dolan MF, Moriya S, DelgadoViscogliosi P, Kudo T, Capron M, Pierce RJ, Ohkuma M, Viscogliosi E.
Molecular Phylogenetic position of the genera Stephanonympha and
Caduceia (Parabasalia) inferred from nuclear small subunit rRNA gene
sequences.
Duboucher C, Barbier C, Beltramini A, Rona M, Ricome JL, Morel G,
Capron M, Pierce RJ, Dei-Cas E, Viscogliosi E.
Pulmonary superinfection by trichomonads in the course of acute
respiratory distress syndrom.
Lung, 2007, 185:295-301.
Peyrin-Biroulet E, Standaert-Vitse A, Branche J, Chamaillard M.
IBD serological panels: facts and perspectives.
Inflamm Bowel Dis, 2007, 12:1561-1566.
Duboucher C, Boggia R, Morel G, Capron M, Pierce RJ, Dei-Cas E,
Viscogliosi E.
Pneumocystis pneumonia : immunodepression, Pneumocystis jirovecii …
and the third man.
Nat Rev Microbiol, 2007, 5:12.
Soltane R, Guyot K, Dei-Cas E, Ayadi A.
Cryptosporidium in calves : results of a longitudinal study in a dairy farm
in Sfax, Tunisia.
Parasite, 2007, 14:309-312.
Dufernez F, Walker RL, Noël C, Caby S, Mantini C, DelgadoViscogliosi P, Ohkuma M, Kudo T, Capron M, Pierce RJ, Villanueva
MR, Viscogliosi E.
Morphological and molecular identification of non-Tritrichomonas fœtus
trichomonad protozoa from the bovine preputial cavity.
Soltane R, Guyot K, Dei-Cas E, Ayadi A.
Prevalence of Cryptosporidium spp. in seven species of farm animals in
Tunisia.
Parasite, 2007, 14:335-338.
Soula F, Fréalle E, Durand-Joly I, Dutoit E, Rouland V, Renard E,
Houfflin-Debarge V, Subtil D, Camus D, Dei-Cas E, Delhaes L.
Intérêt de l’indice d’avidité des IgG pour le diagnostic d’exclusion d’une
toxoplasmose récente: comparaison de la trousse TOXO IgG Avidité SFRI
à la trousse TOXO IgG Avidity Vidas.
Ann Biol Clin (Paris), 2007, 65:257-264.
François N, Fievez A, Standaert-Vitse A, Poulain D, Camus D, Sendid, B.
Prospective evaluation of the new chromogenic medium Candiselect 4
for differentiation and presumptive identification of the mos pathogenic
Candida species.
J Med Microbiol, 2007, 56:495-499.
182
Contents
Kauffmann-Lacroix C, Bousseau A, Dalle F, Brenier-Pinchart MP,
Delhaes L, Machouart M, Gari-Toussaint M, Datry A, Lacroix C,
Hennequin C, Toubas D, Morin O.
Prevention of fungal infections related to the water supply in French hospitals.
Presse Med, 2008, 37:751-759.
Stensvold CR, Suresh KG, Tan KSW, Thompson RCA, Traub RJ,
Viscogliosi E, Yoshikawa H, Clark CG.
Terminology for Blastocystis subtypes – a consensus.
2008
Nevez G, Chabé M, Rabodonirina M, Virmaux M, Dei-Cas E, Hauser
P, Totet A.
Nosocomial Pneumocystis jirovecii infections.
Parasite, 2008, 15:359-365.
Aliouat-Denis CM, Demanche C, Courpon A, Chabé M, Guillot J, DeiCas E, Aliouat EM.
Microchampignons et Chauves-souris.
Symbioses, 2008, 21:74.
Sendid B, Dotan N, Nseir S, Savaux C, Vandewalle P, StandaertVitse A, Zerimech F, Guery BP, Dukler A, Colombel JF, Poulain D.
Antibodies against glucan, chitin, and Saccharomyces cerevisiae mannan
as new biomarkers of Candida albicans infection that complement tests
based on C. albicans mannan.
Aliouat-Denis CM, Chabé M, Demanche C, Aliouat EM, Viscogliosi E,
Guillot J, Delhaes L, Dei-Cas E.
Pneumocystis species, coevolution and pathogenic power.
Boorom K, Smith H, Nimri L, Viscogliosi E, Spanakos G, Parkar U,
Li LH, Zhou XN, Ok U, Jones M, Leelayoova S.
Oh my aching gut : irritable bowel syndrome, Blastocystis, and
asymptomatic infection.
BMC Parasit Vectors, 2008, 1: 40.
Vandewalle P, Colombel JF, Joossens S, Standaert-Vitse A, Collot M,
Halfvarson J, Hadrich I, Landers C, Vermeire S, Rutgeerts P, Targan
S, Mallet JM, Chamaillard M, Sendid B, Poulain D.
Detection of antisynthetic mannoside antibodies (AMA) reveals
heterogeneity in the ASCA response of Crohn's Disease patients and
contributes to differential diagnosis, stratification, and prediction.
Boury D, Dei-Cas E.
Current bioethical issues in parasitology.
Parasite, 2008, 15:489-494.
Vives MF, Caspar-Bauguil S, Aliouat EM, Escamilla R, Perret B,
Dei-Cas E, Prévost MC.
Hypobaric hypoxia-related impairment of pulmonary surfactant proteins
A and D did not favour Pneumocystis carinii Frenkel 1999 growth in nonimmunocompromised rats.
Parasite, 2008, 15:53-64.
Collot M, Sendid B, Fievez A, Savaux C, Standaert-Vitse A,
Tabouret M, Drucbert AS, Danzé PM, Poulain D, Mallet JM.
Biotin sulfone as a new tool for synthetic oligosaccharide immobilization :
application to multiple analysis profiling and surface plasmonic analysis
of anti-Candida albicans antibody reactivity against alpha and beta
(1-->2) oligomannosides. J
Med Chem, 2008, 51:6201-6210.
Yéra H, Zamfir O, Bourcier T, Viscogliosi E, Noël C, Dupouy-Camet J,
Chaumeil C.
Characterization of human Acanthamoeba isolates from ocular samples.
Br J Ophthalmol, 2008, 92:1139-1141.
Dei-Cas E, Dupouy-Camet J.
Foreword.
Parasite, 2008, 15:185-186.
2009
Delhaes L, Harun A, Chen S, Nguyen Q, Maszewska K, Halliday C,
Robert V, Sorrell T, Slavin M, Health C.
Auscedo Study Group TA, Meyer W. Molecular typing of Australian
Scedosporium isolates showing genetic variability and numerous S.
aurantiacum.
Emerg Infect Dis, 2008, 14 :282-290.
Ajzenberg D, Yera H, Marty P, Paris L, Dalle F, Menotti J, Aubert D,
Franck J, Bessières M-H, Quinio D, Pelloux H, Delhaes L, Desbois N,
Thulliez P, Robert-Gangneux F, Kauffman-Lacroix C, Pujol S,
Rabodonirina M, Bougnoux M-E, Cuisenier B, Duhamel C, Duong T-H,
Filisetti D, Flori P, Gay-Andrieux F, Pratlong F, Nevez G, Totet A,
Carme B, Dardé M-L, Villena I.
Genotype of 88 Toxoplasma gondii isolates associated with
toxoplasmosis in immunocompromised patients, and correlation with
clinical findings.
J Infect Dis, 2009, 199:1155-1167.
Duboucher C, Pierce RJ, Capron M, Dei-Cas E, Viscogliosi E.
Recent advances in pulmonary trichomonosis.
Dufernez F, Derelle E, Noël C, Sanciu G, Mantini C, Dive D, SoyerGobillard MO, Capron M, Pierce RJ, Wintjens R, Guillebault D,
Viscogliosi E.
Molecular characterization of iron-containing superoxide dismutases in
the heterotrophic dinoflagellate Crypthecodinium cohnii.
Protist, 2008, 159:223-238.
Aliouat-Denis CM, Martinez A, Aliouat EM, Pottier M, Gantois N,
Dei-Cas E.
The Pneumocystis life cycle.
Mem Inst Oswaldo Cruz, 2009, 104:419-426.
Bachega JFR, Nava MVAS, Bleicher L, Bortoleto-Bugs RK, Dive D,
Hoffmann P, Viscogliosi E, Garratt RC.
Systematic structural studies of superoxide dismutases from human
parasites and a statistical coupling analysis of metal binding specificity.
Proteins, 2009, 77:26-37.
Harry M, Dupont L, Romana C, Demanche C, Mercier A, Livet A,
Diotaiuti L, Noireau F, Emperaire L.
Microsatellite markers in Triatoma pseudomaculata (Hemiptera,
Reduviidae, Triatominae), Chagas' disease vector in Brazil.
Chabe M, Nevez G, Totet A, Frealle E, Delhaes L, Aliouat EM, Dei-Cas E.
Pneumocystis transmission.
J Mycol Med, 2009, 19:276-284.
Jawhara S, Thuru X, Standaert-Vitse A, Jouault T, Sendid B,
Mordon S, Desreumaux P, Poulain D.
Candida albicans colonisation and colonic inflammation induced by
dextran sulphate sodium in wild type and galectin 3-deficient mice.
J Infect Dis, 2008, 197:972-980.
Derouiche S, Deville M, Taylor ML, Akbar H, Guillot J, Pottier M,
Aliouat EM, Aliouat-Denis CM, Dei-Cas E, Demanche C.
Pneumocystis diversity as a phylogeographic tool.
Mem Inst Oswaldo Cruz, 2009, 104 :112-117.
183
Contents
Duarte-Escalante E, Zúñiga G, Ramírez ON, Córdoba S, Refojo N,
Arenas R, Delhaes L, Reyes-Montes Mdel R.
Population structure and diversity of the pathogenic fungus Aspergillus
fumigatus isolated from different sources and geographic origins.
Mem Inst Oswaldo Cruz, 2009, 104 :427-433.
Montes-Cano MA, Chabé M, Fontillon-Alberdi M, de la Horra C,
Respaldiza N, Medrano FJ, Varela JM, Dei-Cas E, Calderon EJ.
First molecular evidence of Pneumocystis jirovecii vertical transmission
in Human.
Emerg Infect Dis, 2009, 15:125-127.
Dupouy-Camet J, Olesen OF, Dei-Cas E, Loiseau PM, Mas-Coma S.
Perspectives for parasitology and parasitology networks in Europe.
Noda S, Mantini C, Bordereau C, Kitade O, Dolan MF, Viscogliosi E.
Ohkuma Molecular phylogeny of parabasalids with emphasis on the
order Cristamonadida and its complex morphological evolution.
Mol Phylogenet Evol, 2009, 52:217-224.
Fréalle E, Decrucq K, Botterel F, Bouchindhomme B, Camus D,
Dei-Cas E, Costa JM, Yakoub-Agha I, Bretagne S, Delhaes L.
Diagnosis of invasive aspergillosis using bronchoalveolar lavage in
haematology patients: Influence of bronchoalveolar lavage human DNA
content on real-time PCR performance.
Eur J Clin Microbiol Infect Dis, 2009, 28:223-232.
Pihet M, Carrere J, Cimon B, Chabasse D, Delhaes L, Symoens F,
Bouchara JP.
Occurrence and relevance of filamentous fungi in respiratory secretions
of patients with cystic fibrosis - a review.
Med Mycol, 2009, 47:387-397.
Grenouillet F, Botterel F, Crouzet J, Larosa F, Hicheri Y, Ranque S,
Delhaes L.
Scedosporium prolificans : an emerging pathogen in France ?
Med Mycol, 2009, 47:343-350.
Poulain D, Sendid B, Standaert-Vitse A, Fradin C, Jouault T,
Jawhara S, Colombel JF.
Yeasts : Neglected Pathogens.
Dig Dis, 2009, 27:104-110.
Heath CH, Slavin MA, Sorrell TC, Handke R, Harun A, Phillips M,
Nguyen Q, Delhaes L, Ellis D, Meyer W, Chen SC; Australian
Scedosporium Study Group.
Population-based surveillance for scedosporiosis in Australia :
epidemiology,disease manifestations and emergence of Scedosporium
aurantiacum infection.
Clin Microbiol Infect, 2009, 15:689-693.
Sendid B, Jouault T, Standaert-Vitse A, Fradin C, Colombel JF,
Poulain D.
Anti-glycan antibodies establish an unexpected link between C. albicans
and Crohn disease.
Med Sci, 2009, 25 :473-481.
Souppart L, Sanci G, Cian A, Wawrzyniak I, Delbac F, Capron M,
Dei-Cas E, Boorom K, Delhaes L, Viscogliosi E.
Molecular epidemiology of human Blastocystis isolates in France.
Parasitol Res, 2009, 105:413-421.
Jiménez JC, Pinon A, Dive D, Grzych JM, Capron M, Di Prisco MC,
Dei-Cas E.
Specific IgA antibody response in children infected with Giardia
intestinalis after treatment with Secnidazol.
Am J Trop Med Hyg, 2009, 80:11-15.
Standaert-Vitse A, Sendid B, Joossens M, François N, VandewalleEl Khoury P, Branche J, Van Kruiningen H, Jouault T, Rutgeerts P,
Gower-Rousseau C, Libersa C, Neut C, Broly F, Chamaillard M,
Vermeire S, Poulain D, Colombel JF.
Candida albicans colonization and ASCA in familial Crohn’s disease.
Jimenez JC, Fontaine J, Grzych JM, Capron M, Dei-Cas E.
Antibody and cytokine responses in BALB/c mice immunized with the
excreted/secreted proteins of Giardia intestinalis : the role of cysteine
proteases.
Ann Trop Med Parasitol, 2009, 103:1-11.
Yera H, Filisetti D, Bastien P, Ancelle T, Thulliez P, Delhaes L.
Multicenter comparative evaluation of five commercial methods for
toxoplasmic DNA extraction in amniotic fluid.
J Clin Microbiol, 2009, 47:3881-3886.
Kaneshiro E, Dei-Cas E.
The 10th International Workshops on Opportunistic Protists (IWOP-10).
Euk Cell, 2009, 8:426-428.
Magnino S, Colin P, Dei-Cas E, Madsen M, Prieto-Maradona M,
McLauchlin J, Nöckler K, Tsigarida K, Vanopdenbosch E,
Van Peteghem C.
Biological risks associated with consumption of reptile products.
2010
ANOFEL
Cryptosporidium National Network. Laboratory-based surveillance for
Cryptosporidium in France, 2006-2009.
Euro Surveill, 2010, 15:19642.
Mantini C, Dalia-Cornette J, Noda S, Van der Heijden HMJF,
Dei-Cas E, Landman WJM, Ohkuma M, Viscogliosi E.
Molecular identification and phylogenetic relationships of trichomonad
isolates of galliform birds inferred from nuclear small subunit rRNA gene
sequences.
Parasitol Res, 2009, 106:163-170.
Borman A, Palmer M, Delhaes L, Horré R, Bouchara JP.
Lack of standardization in the procedures for mycological examination
of sputum samples from CF patients : a possible cause for variations in
the prevalence of filamentous fungi.
Med Mycol, 2010, in press
Calderón E, Dei-Cas E.
Pneumocystis infection : unraveling the colonization-to-disease shift.
Expert Rev Anti Infect Ther, 2010, 8:683-701.
Mantini C, Souppart L, Noël C, Duong TH, Mornet M, Carroger G,
Dupont P, Masseret E, Goustille J, Capron M, Duboucher C,
Dei-Cas E, Viscogliosi E.
Molecular characterization of a new Tetratrichomonas species in empyema.
J Clin Microbiol, 2009, 47:2336-2339.
Calderón E, Gutiérrez-Rivero S, Durand-Joly I, Dei-Cas E.
Pneumocystis infection in humans : diagnosis and treatment.
Expert Rev Anti Infect Ther, 2010, in press.
Martinez A, Aliouat EM, Pottier M, Gantois N, Pinçon C, StandaertVitse A, Dei-Cas E, Aliouat-Denis CM.
High-speed cell sorting of infectious trophic and cystic forms of
Pneumocystis carinii.
184
Contents
Certad G, Creusy C, Ngouanesavanh TM, Guyot K, Gantois N,
Mouray A, Chassat T, Flament N, Fleurisse L, Pinon A, Delhaes L,
Dei-Cas E.
Development of Cryptosporidium parvum induced gastro-intestinal
neoplasia in SCID mice : severity of lesions is correlated with infection
intensity.
Am J Trop Med Hyg, 2010, 82:257-265
Niquil N., Kagami M, Urabe J, Christaki U, Viscogliosi E,
Sime-Ngando T.
Potential role of fungi in plankton food-web functioning and stability : a
simulation analysis based on Lake Biwa inverse model.
Hydrobiologia, 2010, in press.
Roumy V, Hennebelle T, Aliouat EM, Pottier M, Biabiany M, Joha S,
Quesnel B, Khatib R, Sahpaz S, Bailleul F.
Antifungal and cytotoxic activity of withanolides from Acnistus arborescens.
J Nat Prod, 2010, in press.
Certad C, Creusy C, Guyot K, Mouray A, Chassat T, Delaire B,
Pinon A, Sitja-Bobadilla A, Alvarez-Pellitero P, Praet M, Cuvelier C,
Dei-Cas E.
Fulminant Cryptosporidiosis Associated with Digestive Adenocarcinoma
in SCID Mice Infected with /C. parvum/ TUM1.
Int J Parasitol, 2010, in press.
Sarr D, Aldebert D, Marrama L, Frealle E, Gaye A, Brahim HO, Niang M,
Dangou JM, Mercereau-Puijalon O, Lehesran JY, Jambou R.
Chronic infection during placental malaria is associated with up-regulation
of cycloxygenase-2.
Malaria J, 2010, 9:45.
Chabé M., Aliouat-Denis CM, Delhaes L, Aliouat EM, Viscogliosi E,
Dei-Cas E.
Pneumocystis : from an uncertain unique entity to a group of highly
diversified fungal species.
FEMS Yeast Res, 2010, in press.
Mekinian A, Queyrel V, Durand-Joly I, Moranne O, Denis G, Delhaes L,
Morell-Dubois S, Lambert M, Launay D, Hachulla E, Hatron PY.
Positive
Pneumocystis jirovecii PCR in immunocompromised patients with a
systemic disease : Infection or colonisation ?
Rev Med Interne, 2010, in press.
Chabé M, Herbreteau V, Hugot JP, Bouzard N, Deruyter L,
Morand S, Dei-Cas E..
Pneumocystis carinii and Pneumocystis wakefieldiae in Wild Rattus
norvegicus Trapped in Thailand.
Souppart L, Moussa H, Cian A, Sanciu G, Poirier P, El Alaoui H,
Delbac F, Boorom K, Delhaes L, Dei-Cas E, Viscogliosi E.
Subtype analysis of Blastocystis isolates from symptomatic patients in
Egypt.
Parasitol Res, 2010, 106:505-511.
Chabé M, Herbreteau V, Hugot JP, Chaval Y, Bouzard N, Deruyter L,
Morand S, Dei-Cas E.
Pneumocystis spp in wild South East murine rodents.
J Vet Kasetsart University, 2010, in press.
Sterkers Y, Varlet-Marie E, Cassaing S, Brenier-Pinchart MP, Brun S,
Dalle F, Delhaes L, Filisetti D, Pelloux H, Yera H, Bastien P.
Multicentric comparative analytical performance study for molecular
detection of low amounts of Toxoplasma gondii from simulated
specimens.
J Clin Microbiol, 2010, 48:3216-3222.
Dei-Cas E.
Ethique du vivant et épistémologie de la biologie.
Ethique et Santé, 2010, in press.
Delhaes L, Fréalle E, Pinel C.
Serodiagnosis of Aspergillus infections in patients with cystic fibrosis.
Med Mycol, 2010, in press.
Villena I, Ancelle T, Delmas C, Garcia P, Brezin AP, Thulliez P,
Wallon M, King L, Goulet V.
Toxosurv network and National Reference Centre for Toxoplasmosis.
Congenital toxoplasmosis in France in 2007: first results from a national
surveillance system.
Euro Surveill, 2010, 15:pii:19600.
Delhaes L, Ajzenberg D, Sicot B, Bourgeot P, Dardé ML, Dei-Cas E,
Houfflin-Debarge V.
Severe congenital toxoplasmosis due to a Toxoplasma gondii strain with
an atypical genotype: case report and review.
Prenatal Diag, 2010, 30:902-905.
Book Chapter
Delhaes L, Mraz JC, Fréalle E, Durand-Joly I, Magro L, Ajzenberg D,
Dardé ML, Dei-Cas E, Yakoub-Agha I.
Severe pulmonary toxoplasmosis after allogenic stem cell transplantation
in two patients : from Toxoplasma genotyping to clinical management.
Bone Marrow Transplant, 2010, in press.
2008
Dei-Cas E, Chabé M, Durand-Joly I, Aliouat-Denis CM, Aliouat EM.
Pneumocystis y Pneumocystosis.
In : « Actualidades en Micologia Médica », 4th edition, Méndez-Tovar
LJ, Lopez Martinez R, Hernandez Hernandez F (eds.), Faculté de
Médecine, UNAM, Mexico DF, 2008, pp 287-295.
Houssin T, Follet J, Follet A, Dei-Cas E, Senez V.
Label-Free Analysis of Water-Polluting Parasite by Electrochemical
Impedance Spectroscopy.
Biosens Bioelectron, 2010, 25:1122-1129.
2009
Laurent J, Stanicki D, Huang TL, Dei-Cas E, Pottier M, Aliouat EM,
Vanden Eynde JJ.
Bisbenzamidines as antifungal agents. Are both amidines functions required
to observe an anti-Pneumocystis carinii activity ?
Molecules, 2010, 15:4283-4293.
Duboucher C, Dei-Cas E, Viscogliosi E.
On the finding of trichomonads as co-infecting agents in Pneumocystis
pneumonia.
In : « AIDS-Related Opportunistic Infections», Galanda CD (ed.), Nova
Biomedical, Nova Science Publishers, New York, 2009, pp. 81-92.
Mekinian A, Durand-Joly I, Hatron PY, Moranne O, Denis G, Dei-Cas E,
Morell-Dubois S, Lambert M, Launay D, Delhaes L, Hachulla E,
Queyrel V.
Pneumocystis jirovecii colonization in patients with systemic autoimmune
diseases : Prevalence, Risk Factors of Colonization and Outcome.
Rheumatology, 2010, in press.
185
Contents
2010
HDR
Chabé M, Hugot JP, Dei-Cas E.
Pneumocystis molecular phylogeny : a way to understand both
pneumocystosis natural history and host taxonomy.
In : «New frontiers of Molecular Epidemiology of Infectious Diseases»,
Morand S, Beaudeau F, Cabaret J, de Rycke J (eds), 2010, in press.
Meja RABODONIRINA
Tuteur : Eduardo Dei-Cas
« Mycoses and parasitoses associated with HIV/AIDS infection: descriptive
and molecular epidemiology”
Université de Lyon 1
18 Décembre 2007
Creusy C, Certad G, Guyot K, Dei-Cas E.
Parasites and oncogenesis with a special reference to gastro-intestinal
neoplasia induced by Cryptosporidium parvum.
In : «Detection of Bacteria, Viruses, Parasites and Fungi», M. ViolaMagni (ed.), NATO Series Science for Peace, Springer, Berlin, 2010,
in press.
Laurence DELHAES
Tuteur : Eduardo Dei-Cas
20 Décembre 2007
Dei-Cas E, Verrez-Bagnis V.
Les parasites des poissons.
In : «Les produits aquatiques : caractéristiques, transformation,
valorisation». Éditions Lavoisier, Paris, 2010, in press.
Dei-Cas E, Aliouat CM, Certad G, Creusy C, Guyot K.
Infectious forms of parasites in food : man embedded in ecosystems.
In : «Detection of Bacteria, Viruses, Parasites and Fungi», M. ViolaMagni (ed.), NATO Series Science for Peace, Springer, Berlin, 2010, in
press.
Singahl A, Aliouat EM, Creusy C, Kaplan G, Bifani P.
Histopathological manisfestation in pulmonary tuberculosis in rats. A Color
Atlas of Comparative Pathology of Pulmonary Tuberculosis.
Ed. Leong J, Dartois V and Dick T. Taylor and Francis. 2010, 128-157.
PhD
Christophe DUBOUCHER
Directeur de thèse : Eric Viscogliosi
“ Contribution to the study of pulmonary trichomonosis”
Doctorat avec option éventuelle : Biologie-Parasitologie
14 Décembre 2007
Tra My NGOUANESAVANH
Directeur de thèse : Jean-Charles Cailliez
« Molecular epidemiology of cryptosporidiosis »
18 Décembre 2007
Gabriela CERTAD
Directeur de thèse : Eduardo Dei-Cas
« Genetic characterization of Cryptosporidium an dits implication in
digestive neoplasia”
30 Septembre 2008
Emilie FREALLE
Directeur de thèse : Laurence Delhaes
“ Molecular characterization of mould antioxidative system an dits
physiopathogenic relevance during Aspergillus fumigatus diseases”
21 Octobre 2008
186
Contents
Contents
Technological
Facilities
188
Contents
Plethysmography/
Functional Investigation
of Airway Diseases
Inserm U 1011
David Dombrowicz
Contact :
00 33 3 20 87 79 67
[email protected]
Group members :
Dombrowicz David, DR Inserm (team 3, U 1011)
Duez Catherine, CR1 Inserm (team 11, U 1019)
Fleury Sébastien, AI Inserm (team 3, U 1011)
Marquillies Philippe, Technician IPL (team 11, U 1019)
189
Contents
1) Mission of the platform
Publications
To provide scientists from the Pasteur Institute, as well as
researchers from academia and industry with up-to date tools
for the investigation of airway function.
2007
IL-18 Does not Increase Allergic Airway Disease in Mice When Produced
by BCG.
2) Description of the platform
J Biomed Biotechnol, 2007, 67276.
Plethysmography plateform is equipped for non-invasive and
invasive measurements of lung function.
2008
Non invasive plethysmography.
Plateform is equipped for whole-body plethsymography on
conscious unrestrained animals with a Buxco-type system (EMKA,
France) (left) allowing for the simultaneous analysis of 8 mice.
Longitudinal studies of airway hyperreactivity are thus possible.
System is suitable for mice on a Balb/c background and is particularly appropriate for pre-clinical evaluation of a large number
of compounds. Analysis of up to 40 mice per day is possible.
Verhasselt V, Milcent V, Cazareth J, Kanda A, Fleury S,
Dombrowicz D, Glaichenhaus N and Julia V.
Breast milk-mediated transfer of an antigen induces tolerance and
protection from allergic asthma.
Nat Med, 2008, 14:170-175.
2009
Legrand F, Staumont-Salle D, Queant S, Bertout J, Fleury S, Remy P,
Papin JP, Julia V, Capron M and Dombrowicz D.
Eosinophil-derived IFN-gamma induces airway hyperresponsiveness and
lung inflammation in the absence of lymphocytes.
J Allergy Clin Immunol, 2009, 124:573-582, 582 e571-579.
Invasive plethysmography.
Plateform is equipped for invasive plethysmography
on anesthetized animals with a Flexivent System (Scireq,
Canada) (right). Using various mathematical models (single
compartment, forced oscillations, constant phase, PV-loops),
this device allows for detailled investigation in mice of all the
parameters of respiratory mechanics used in humans such as
resistance, compliance, elastance, impedance... System is
suitable for detailled studies regardless of the mouse genetic
background. Analysis of up to 12 mice per day is possible.
Plethysmography plateform is run by D. Dombrowicz (Team 3,
Inserm U1011) in association with C. Duez (Team 11, Inserm
U1019) with the technical help form S. Fleury (U1011) and
P. Marquillies (U1019).
2010
Dubois A, Deruytter N, Adams B, Kanda A, Delbauve S, Fleury S,
Torres D, François A, Pétein M, Goldman M, Dombrowicz D, Flamand V.
Regulation of Th2 responses and allergic inflammation through
bystander activation of CD8+ T lymphocytes in early life.
J Immunol, 2010, 185:883-891.
Mionnet C, Buatois V, Kanda A, Cossalter G, Milcent V, Fleury S,
Hessel E, Coffman RL, Dombrowicz D, Glaichenhaus N, and Julia V.
CX3CR1 is required for allergic airway inflammation by promoting Th2
cell survival and maintenance in inflamed lung.
Nat Med, 2010, in press.
3) Projects
Most investigations sofar were dedicated to allergic diseases
(allergic asthma or atopic dermatitis). Typical studies involve
either the comparison of airway function in geneticallymodified (transgenic or "knock-out") and wild type mice or
study of the efficiency of immunomodulatory compounds
(cytokines, chemo-kines, nuclear or prostaglandin receptor
agonists...).
Both non-invasive and invasive systems can also be used to
monitor airway function in other physiopatholgical conditions
(bacterial or viral lung infections, stress, obesity....).
Current projects using the plateform aim to investigate the
role of NK cells and eosinophils in allergic asthma and the
regulation of the pathology through nuclear receptors.
Mosconi E, Rekima A, Seitz-Polski B, Kanda A, Fleury S, Tissandie E,
Monteiro R, Dombrowicz D, Julia V, Glaichenhaus N, Verhasselt V.
Breast milk immune complexes are potent inducers of oral tolerance in
neonates and prevent asthma development.
Mucosal Immunol, 2010, 3:461-474.
Navarro S, Cossalter G, Chivaroli C, Kanda A, Fleury S, Cazareth J,
Sparwasser T, Dombrowicz D, Glaichenhaus N and Julia V.
The oral administration of bacterial extracts prevents asthma via the
recruitment of regulatory T cells to the airways.
Mucosal Immunol, 2010, in press.
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Microscopy - Imaging Cytometry of
the Pasteur Lille Campus
(MICPaL) facility
Inserm U 1019 - CNRS UMR 8204
Frank LAFONT
Contact :
00 33 3 20 87 11 36
[email protected]
Group members :
Lafont Frank, DR2 CNRS
Barois Nicolas, IR Inserm
Bertout Julie, CE IPL
Bongiovanni Antonino, Technician IPL
Faveeuw Christelle, CRI Inserm
Janel Sébastien, IE CNRS
Werkmeister Elisabeth, IR CNRS
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1) Mission of the Facility
Publications
The Microscopy-Imaging-Cytometry facility core unit of the
Pasteur Lille campus (MICPaL Facility) was created in 2006 by
the joined effort of the 4 preexisting technical facilities for flux
cytometry electron microscopy, biophotonic microscopy and
atomic force microscopy. The MICPaL facility is engaged
towards ISO 9001 quality assessment certification. The Facility
is involved in local, national and international trainings
(lectures and workshops,…). The MICPaL Facility is member of
the Molecular and Cellular Medicine Federative Research
Institute (IFR142).
2007
Foligne B, Zoumpopoulou G, Dewulf J, Ben Younes A, Chareyre F,
Sirard JC, Pot B, Grangette C.
A key role of dendritic cells in probiotic functionality.
PLoS One, 2007, 2:e313.
Khayath N, Vicogne J, Ahier A, BenYounes A, Konrad C, Trolet J,
Viscogliosi E, Brehm K, Dissous
C FEBS J, 2007, 274:659-676.
The Facility is :
- open to the Research Groups hosted on the campus,
- open locally to the scientific community,
- open to the national and international scientific
communities,
- member of national technology networks.
Paget C, Mallevaey T, Speak AO, Torres D, Fontaine J, Sheehan KC,
Capron M, Ryffel B, Faveeuw C, Leite de Moraes M, Platt F,
Trottein F.
Immunity, 2007, 27:597-609.
Mallevaey T, Fontaine J, Breuilh L, Paget C, Castro-Keller A,
Vendeville C, Capron M, Leite-de-Moraes M, Trottein F, Faveeuw C.
Invariant and noninvariant natural killer T cells exert opposite regulatory
functions on the immune response during murine schistosomiasis.
Infect Immunity, 2007, 75:2171-2180
The MICPaL facility is engaged in partnerships with industries
leader in Microscopy techniques.
2) Description of the Facility
Spriet C, Trinel D, Waharte F, Deslee D, Vandenbunder B, Barbillat J,
Héliot L.
Correlated fluorescence lifetime and spectral measurements in living cells.
Microsc Res Tech, 2007, 70:85-94.
cf. www.micpal.fr
3) Implicated projects
The scientific projects in which the MICPaL facility in involved
cover the main themes developed on the campus : infectionimmunity, cancer and metabolic diseases and the personal
from the facility collaborates with scientists located outside
the campus. It also provides services for academics and
industries.
Trolet J, Fafeur V, Ben Younes A, Dissous C
Int J Parasitol, 2007, 37:1539-1550.
Yersin A, Hirling H, Kasas S, Roduit C, Kulangara K, Dietler G,
Lafont F, Catsicas S, Steiner P.
Elastic properties of the cell surface and trafficking of single AMPA
receptors in living hippocampal neurons.
Biophys J, 2007, 92:4482-4489.
2008
Richard A, Barras A, Younes AB, Monfilliette-Dupont N, Melnyk P.
Minimal chemical modification of reductive end of dextran to produce
an amphiphilic polysaccharide able to incorporate onto lipid
nanocapsules.
Bioconjug Chem, 2008, 19:1491-1495.
Roduit C, van der Goot FG, De Los Rios P, Yersin A, Steiner P,
Dietler G, Catsicas S, Lafont F, Kasas S.
Elastic membrane heterogeneity of living cells revealed by stiff nanoscale
membrane domains.
Biophys J, 2008, 94:1521-1532.
Torres D, Paget C, Fontaine J, Mallevaey T, Matsuoka T, Maruyama T,
Narumiya S, Capron M, Gosset P, Faveeuw C, Trottein F.
Prostaglandin D2 inhibits de production of IFN-gamma by invariant NK
T cells : consequences in the control of B16 melanoma.
J Immunol, 2008, 180:738-792.
2009
Bialecki E, Paget C, Fontaine J, Capron M, Trottein F, Faveeuw, C.
Role of marginale zone B lymphocytes in invariant NKT cell activation.
J Immunol, 2009, 182:6105-6113.
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Dupont N, Lacas-Gervais S, Bertout J, Paz I, Freche B, Van Nhieu GT,
van der Goot FG, Sansonetti PJ, Lafont F.
Shigella phagocytic vacuolar membrane remnants participate in the
cellular response to pathogen invasion and are regulated by autophagy.
Cell Host Microbe, 2009, 6:137-149.
Dupres V, Verbelen C, Raze D, Lafont F, Dufrêne YF.
Force spectroscopy of the interaction between mycobacterial adhesins
and heparan sulphate proteoglycan receptors.
Chemphyschem, 2009, 10:1672-1675.
Am J Pathol, 2009, 174:423-435.
Legrand F, Staumont-Sallé D, Quéant S, Bertout J, Fleury S, Rémy P,
Papin JP, Julia V, Capron M, Dombrowicz D.
Eosinophil-derived IFN-gamma induces airway hyperresponsiveness and
lung inflammation in the absence of lymphocytes.
J Allergy Clin Immunol, 2009, Jun 17.
Paget C, Bialecki E, Fontaine J, Vendeville C, Mallevaey T, Faveeuw
C, Trottein F.
Role of invariant NK T lymphocytes in immune responses to CpG oligo
deoxynucleotides
J Immunol, 2009, 18:1846-1853.
Roduit C, Sekatski S, Dietler G, Catsicas S, Lafont F, Kasas S.
Stiffness tomography by atomic force microscopy.
Biophys J, 2009, 97:674-677.
2010
Feunou PF, Bertout J, Locht C.
T- and B-cell-mediated protection induced by novel, live attenuated
pertussis vaccine in mice. Cross protection against parapertussis.
PLoS One, 2010, 5:e10178.
Ple C, Barrier M, Amniai L, Marquillies P, Bertout J, Tsicopoulos A, Walzer
T, Lassalle P, Duez C.
Natural killer cells accumulate in lung-draining lymph nodes and regulate
airway eosinophilia in a murine model of asthma.
Scand J Immunol, 2010, 72:118-27.
Janot L, Erard F, Bertout J, Leger H, Sebbane F, Benecke A, Renauld JC,
Hardt WD, Ryffel B, Sirard JC.
TLR5 signaling stimulates the innate production of IL-17 and IL-22 by
CD3(neg)CD127+ immune cells in spleen and mucosa.
J Immunol, 2010, 185:1177-85.
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Surface Plasmon
Resonance Platform
CNRS UMR 8161
Marc AUMERCIER
Contact :
00 33 3 20 87 11 34
[email protected]
Group members :
Aumercier Marc, CRI CNRS
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- Mons-Hainaut University, Belgium :
Pr Robert Muller
1) Mission :
Study of contrast agents used in NMR.
The Biacore® 2000 apparatus using Surface Plasmon
Resonance (SPR) is dedicated to the study of the biomolecular
interactions. It enables to characterize the different kinetic
parameters or the different parameters at the equilibrium of
the interaction (kon, koff, KD and KA). It makes possible the
detection and quantification of molecules interacting with
each other. Analysis of numerous interaction types can be
performed: protein-protein, DNA-protein, DNA-RNA, sugarprotein, lipid-protein or every type of molecules used in
biomedical research. The study of particle-molecule or even
cell-molecule interactions can also be done with this
apparatus. Characterization of interactions can be done from
either purified molecules or cellular lysates and growth
mediums.
This instrument is accessible for research projects coming
from the Pasteur Institute of Lille as well from regional and
national laboratories. It has also a training vocation. Indeed,
the Pasteur Institute of Lille School of Training organizes a
training session called “Introduction to the Biacore®
Technology” including a sizeable experimental part which is
done using the Biacore® 2000.
- CEA Grenoble :
Dr Victor Duarte
Study of the bacterial transcription factor PerR.
- Lens University :
Pr Didier Betbeder
Characterization of nanoparticles for drug vectorization.
Publications
2008
Laitem C, Choul-Li S, Baillat D, A. Bègue A, Aumercier M.
Efficient system for biotinylated recombinant Ets-1 production in
Escherichia coli : a useful tool for studying interactions between Ets-1 and
its partners.
Protein Expression and Purification, 2008, 62:53-63.
2009
Baillat D, Laitem C, Leprivier G, C. Margerin C, Aumercier M.
Ets-1 binds cooperatively to the palindromic Ets-binding sites in the p53
promoter.
Bioch Biophys Res Commun, 2009, 378:213-217.
2) Description :
The Surface Plasmon Resonance platform is constituted of a
Biacore® 2000 apparatus based on the biosensor principle.
This machine is completely thermostated and monitored by
computer. It is essentially composed of 1)- an automated
system of fluid transfer, 2)- a gold chip surface (plasmon)
where the ligand is bound, 3)- a microfluidic system within
which the analyte of which we want to study the interaction
with the ligand is injected and 4)- a Surface Plasmon
Resonance detection system. This apparatus has a flow cell
divided into four channels, permitting the study of four
simultaneous interactions. The Biacore® software is in charge
of controlling the machine (BIAcore control), doing kinetic
analysis of the studied interaction (BIAevaluation), and
allowing simulation of various binding situations
(BIAsimulation). In addition, operation of the apparatus can
be controlled by computer through programmable methods
which can be written by users. In this way, the Biacore® 2000
apparatus is able to work continuously and therefore to be
used as a screening tool.
Laitem C, Leprivier G, Choul-Li S, Bègue A, Monte D, Dumont P,
Larsimont D, Duterque-Coquillaud M, And Aumercier M.
Ets-1 p27 : a novel Ets-1 isoform with dominant negative effects on the
transcriptional properties and the subcellular localization of Ets-1 p51.
Oncogene, 2009, 28:2087-2099.
Leroux F, Willery E, Mathys V, Déprez-Poulain R, Delcroix G,
Frenois F, Aumercier M, Locht C,Villeret V, Déprez B, Baulard AR.
Nature Med, 2009,15:537-544.
3) Collaborative work :
- INSERM U629, Pasteur Institute of Lille :
Dr Alain Baulard, Dr Camille Locht
Study of EthR binding to its operator.
- INSERM U761, Pasteur Institute of Lille :
Pr Benoît Déprez
Screening of compounds which are able to potentiate ethionamide
activity.
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Transcriptomics
and Applied Genomics
Group (TAG)
(Inserm U 1019, CNRS UMR 8204
IFR 142
Yves LEMOINE
Contact :
00 33 3 20 87 79 12
[email protected]
Group members :
Lemoine Yves, PU1 Univ Lille 1
Hot David, CR IPL
Blervaque Renaud, IE IPL
Huot Ludovic, CE1 IPL
Slupek-Verpoort Stéphanie, Technician IPL
Lozano Luce, PhD student, Lille 2
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The laboratory of Transcriptomics and Applied Genomics
includes the microarray facilities of the Institut Pasteur de Lille.
This platform is devoted to the realization of genomic projects
as provider or in collaboration (14 publications). The TAG team
was initially funded by the Institut Pasteur de Lille and the
European InterReg II program. The team belongs to the Centre
for Infection and Immunity of Lille. The principal aims of the
laboratory were first to establish a team able to master the
microarray main applications in order to collaborate with
laboratories interested in applying transcriptomic or genomic
studies to their research field, and secondly to train students
and professionals in microarray protocols and applications.
More recently, the team developed its own research interest
in bacterial genomics and transcriptomics.
Laboratory description and skills
Today, the TAG laboratory is able to offer transcriptomic and
comparative genomic solutions for high throughput
approaches, using either slides developed in situ or
commercially purchased. This means that we have developed
(i) bioinformatic solutions to be able to work with and analyze
whole genome sequences and associated data banks for the
optimal design of oligonucleotide reporters, (ii) technical
skills to perform high quality spotting (using Genetix QArray2
spotting robot) with relevant negative and positive
controls, (iii) protocols to extract and purify high quality total
RNAs and subsequent evaluation of their integrity number, as
well as (iv) protocols to synthesize fluorescent probes and to
hybridize them onto our home-made or commercially
purchased slides (Agilent Technologies, Illumina, …).
Alongside these technical aspects a range of scanning (using
InnoScan700 from Innopsys), treatment and analysis
protocols, using mainly Bioconductor packages in the
statistical language R, were established to deal with the
generated data. Finally, for an effective management of the
information generated during all the different steps and to
make sure that our data are MIAME compliant, a local
database (BASE ver2.0) was installed and adapted to our
needs.
Activities and developed skills of our microarray facility are
currently evolving toward the use of deep sequencing
allowing new genomic/transcriptomic applications, among
which transcriptome analysis, transcription factor binding
analysis (ChIP-seq), methylation analysis, SNPs & InDel analysis,
genotyping/GWAS. Some of our projects take advantage of
these new approaches. This technical evolution is happening
together with a re-organization of the different actors in
genomics and transcriptomics in Lille leading to the creation
of a network. In addition to the clustering of equipments, this
network should allow a sharing of experiments and skills in
the field. This network (LIGAN acronym for Lille Genomic
Advanced Network) has obtained the “IBiSA” certification in
September 2009.
Examples of projects :
We are offering complete solutions of genomics going from
the design of molecular tools till the analysis and data mining
in the fields of Gene Expression, Gene Discovery, Genotyping,
DNA Binding Protein Analysis, Epigenetics.
Our expertise was concretized by several local, national or
international collaborations on various themes. For instance :
- we developed microarrays representing the whole genome
of the causative agent of whooping cough, Bordetella
pertussis. In collaboration with Camille Locht’s team (Inserm
U1019), this tool was engaged in transcriptomic studies to
investigate the regulon of the two-component system
BvgAS. This same array was also used, in collaboration with
Nicole Guiso (Institut Pasteur, Paris) to identify genomic
variations of circulating and vaccine strains. More recently,
the expertise developed on B. pertussis was exploited further
to develop, in collaboration with Prof. Roy Gross (University
of Würzburg, Germany) a microarray representing the
genome of Bordetella petrii, an environmental strain of this
bacterial family,
- an oligonucleotide microarray representing genes located
within human chromosomal regions defined by genome
scan studies in Alzheimer’s disease was designed in
collaboration with Jean-Charles Lambert and Philippe
Amouyel (Inserm U744, Institut Pasteur de Lille). This tool
allowed us to compare the transcriptomic profiles in brain
tissues of 12 Alzheimer’s disease patients versus control
brains and to identify genes potentially implicated in the
disease. One of these, a transcription factor, is currently
further studied to determine its binding targets on the
genome using a ChIP-seq approach,
- a transcriptomic analysis on rat, done in collaboration with
Jamal Khalife (Inserm U1019), helped to understand
mechanisms involved in age-dependent protection against
malaria,
- target genes of the Erg protein, belonging to the Ets family
transcription factors and involved in skeleton aging, were
identified in collaboration with M. Duterque (UMR8161),
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- the genomic methylated patterns in drug-treated mouse
were detected using a ChIP-chip approach (academic
collaboration),
- genome wide association studies (GWAS) were conducted
on cattle breeders (industrial partnership),
- our own research project deals with the genomics of
Bordetella pertussis virulence. A first study allowed
identifying 14 new small RNAs in the bacterial genome
using bioinformatic predictions. Some of these small RNAs
are further characterized in order to investigate their
potential implication in the bacterial virulence regulation.
A RNA-seq approach using high through-put sequencing
technologies is in progress to enlarge the catalog of
identified small RNAs.
Publications
2008
Feunou PF, Ismaili J, Debrie AS, Huot L, Hot D, Raze D, Lemoine Y,
Locht C.
Genetic stability of the live attenuated Bordetella pertussis vaccine
candidate BPZE1.
Vaccine, 26(45): 5722-5727.
Bouchez V, Caro V, Levillain E, Guigon G, Guiso N.
Genomic content of Bordetella pertussis clinical isolates circulating in
areas of intensive children vaccination.
PLoS One, Jun 18;3:e2437.
2009
Lechner M, Schmitt K, Bauer S, Hot D, Hubans C, Levillain E,
Locht C, Lemoine Y, Gross R.
Genomic island excisions in Bordetella petrii.
BMC Microbiology, 9:141.
Hauw JJ, Dekosky ST, Lemoine Y, Iwatsubo T, Wavrant-Devrièze F,
Dartigues JF, Tzourio C, Buée L, Pasquier F, Berr C, Mann D,
Lendon C, Alpérovitch A., Kamboh MI, Amouyel P, Lambert JC.
Transcriptomic and genetic studies identify IL-33 as a candidate gene for
Mol Psychiatry, 14:1004-1016.
Bensemain F, Hot D, Ferreira S, Dumont J, Bombois S, Maurage CA,
Hauw JJ, Schraen S, Lemoine Y, Buée L, Berr C, Mann D, Pasquier F,
Mol Psychiatry, 14(1):106-16
2010
Drobecq H, Lemoine Y, Koussa M, Amouyel P, Pinet F.
transcriptomic, proteomic, and antibody protein array study.
J Proteome Res, 2010, 9:3720-3729.
Lucau-Danila A, Laborde L, Legrand S, Huot L, Hot D, Lemoine Y,
Hilbert JL, Hawkins S, Quillet MC, Hendriks T, Blervacq AS.
Identification of novel genes potentially involved in somatic
embryogenesis in chicory (Cichorium intybus L.).
BMC Plant Biol, 2010, 10:122
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Peptide Chemistry,
Systems, Biology
CNRS UMR 8161
Oleg MELNYK
Contact :
00 33 3 20 87 12 14
[email protected]
Group Members :
Melnyk Oleg, DR2 CNRS
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The Peptide CSB platform “Peptide Chemistry, Systems,
Biology” is located in the Institute of Biology, which is a CNRS
institute hosted by the Pasteur Institute of Lille (France). This
platform is headed by Dr Oleg Melnyk (UMR 8161). Most of the
equipment were financed by la Région Nord Pas de Calais, the
CNRS, l’Institut Pasteur de Lille and the Ministry of Research.
The originality of this platform is to combine three
complementary areas of expertise.
The first expertise is peptide and ligation chemistry (Hervé
Drobecq, Nathalie Ollivier, Annick Blanpain, Emmanuelle Boll,
Oleg Melnyk)
Our research activity in this field is focused on the discovery
of novel chemical methods for the convergent synthesis of
complex molecular architectures or proteins. These methods,
which are called ligations in the field of biomolecules and in
particular peptide chemistry, allow the site-specific
chemoselective ligation of fully deprotected fragments. In the
past, we have developed mainly non-native ligation methods,
i.e. methods that produce non-native bonds such as
hydrazone or semicarbazone bonds between the ligated
fragments.
Our main goal today is to find chemical methods permitting
the formation of native peptide bonds between two fully
deprotected peptides. These methods are used for the total
synthesis of proteins of moderate size (< 200 amino acids) that
cannot be produced by conventional chemical or
recombinant techniques, or that must be produced without
potential biological contaminants.
Fig. 1. Polystyrene 96-well plates were chemically modified to allow the
site-specific immoibilization of peptides. In this figure, model HA peptide
was printing in triplicate at different concentrations in the bottom of a
polystyrene 96-well plate using a pin contact microarrayer. The
microarrays were incubated with a murine anti-HA antibody and then
with a goat anti-murine antibody labelled with tetramethylrhodamine.
Detection was performed at 532 nm using a Tecan microarray scanner.
Fig. 2 shows peptide microarrays prepared on polycarbonate. The
widespread use of microarrays for diagnostic applications requires the
development of safe systems. Because glass can break and cut, the
analysis of potentially contaminating samples with glass-based devices
must be avoided. Preparation of microarrays in the bottom of polystyrene
96-well plates is a solution to this problem. Bisphenol A polycarbonate is
also emerging as an interesting material in this field: it is shock-resistant,
it can be easily molded and eliminated by incineration. This polymer is
also used for compact disc manufacturing and the use of this substrate
as a platform for high throughput binding assays has attracted much
attention recently. Unfortunately, only few chemical methods are
available for linking molecules to PC. In this work, we have prepared
peptide microarrays on PC substrates by printing 30 nm silica
nanoparticles derivatized by peptide molecules. The nanoparticles
adhere to the PC substrate and are not removed during the assay. In Fig.
2, we have used again model HA and FLAG peptides to validate the
technology. The methodology was used also for characterizing the
binding properties of antibodies directed against Ab1-42 peptide (see our
paper Souplet, V. et al. Bioconjugate Chem. 2009, 20(3), 550-557).
The design and use of peptide-based systems such as
microarrays constitute a second expertise (Rémi Desmet, JeanPhilippe Ebran, Oleg Melnyk)
The lab is deeply involved for several years in the development
of novel surface chemistries, novel biochemical assays and
novel detection techniques that are implemented in micronano systems. Frequently, peptide chemistry is involved in the
preparation of these systems, but since two years we are
involved in the design of oligo or polysaccharide-based
systems too. Depending on their final use, the microarrays or
micro devices require appropriate substrates, probes and/or
detection techniques (fluorescence, colorimetric, electric…).
Today, microarrays are mainly used in the lab for the screening
of chemical libraries and the study of biomolecular
interactions.
Fig. 1 and 2 illustrate some of our contributions in the field of
surface functionalization.
Fig. 1 shows a peptide microarray prepared in the bottom of
polystyrene 96-well plates. Polystyrene by itself does not allow
the deposition of aqueous solutions using standard
microarrayers and the efficient immobilization of peptides. We
have designed a novel polystyrene functionalization method
that is cheap , easy to implement and which allows the
preparation of high quality peptide microarrays. All the
incubations and washing steps can be performed using
standard equipments for 96-well plates.
Fig. 2. Peptides were printed at 10 µM
in DMF/pH 5.5 5 mM sodium acetate
buffer : 1/1 v/v mixture (final nanoparticle concentration 1% w/v, 3
replicates) on PC slides (75 × 25 × 1
mm). The microarrays were incubated
with Ab-HA, Ab-FLAG or murine IgG (10
µg/mL) or the buffer during 1h at 37°C
and then with tetramethylrhodaminelabelled anti-murine antibodies.
Incubations and washings were
performed using standard 96-well
plates incubation and washing
stations. Detection was performed at
532 nm.
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The third field of expertise is the design of cellular/
biochemical assays related to signal transduction by RTKs
(Jérome Vicogne, Gautier Goormachtigh, Myriam Delatre,
Caroline de Witte, Roland Bourette, Véronique Fafeur).
Publications
We are deciphering signal transduction by tyrosine kinase
receptors (RTKs), and in particular by the MET tyrosine kinase
receptor and its ligand HGF/SF. We have strong expertise in
biochemical and biological assays aimed to examine signal
transduction by RTKs. These assays include enzymatic assays,
including cell based-RTK assays, and biological assays
(apoptosis, proliferation, motility and morphogenesis as
illustrated in Fig. 3). All these assays are described in our
publications since 1995.
Biet F, Marques MAD, Grayon M et al.
Mycobacterium smegmatis produces an HBHA homologue which is not
involved in epithelial adherence.
Microb Infect, 2007, 9:175-182.
2007
Carion O, Souplet V, Olivier C et al.
Chemical micropatterning of polycarbonate for site-specific peptide
immobilization and biomolecular interactions.
Chembiochem, 2007, 8:315-322.
Coffinier Y, Szunerits S, Jama C et al.
Peptide immobilization on amine-terminated boron-doped diamond
surfaces.
Langmuir, 2007a, 23:4494-4497.
Coffinier Y, Szunerits S, Marcus B et al.
Covalent linking of peptides onto oxygen-terminated boron-doped
diamond surfaces.
Diamond Related Materials, 2007b, 16:892-898.
Dezitter X, Hammoudi F, Belverge N et al.
Proteomics unveil corticoid-induced S100A11 shuttling in keratinocyte
differentiation.
Dubs P, Bourel-Bonnet L, Subra G et al.
Parallel synthesis of a lipopeptide library by hydrazone-based chemical
Ligation.
J Combinat Chem, 2007, 9:973-981.
Branching morphogenesis by HGF/SF
We are developing also novel methodologies to quantify
cellular enzymatic activities. In particular, we are developing a
cell-based RTK assays using the AlphaScreen® technology and
an EnSpire Multilabel Plate Reader plate reader. The main
advantage of the AlphaScreen® technology is the possibility to
perform a quantitative signal transduction analysis in a
homogeneous assay.
Other enzymatic assays can be developed as necessary, and
they do not necessarily involve specific equipments. For
example, heparanase is an endo-β-D-glucuronidase that
catalyzes the hydrolytic cleavage of the β-1,4-glycosidic bond
between a D-glucuronate and a D-glucosamine in heparan
sulphate. This enzyme plays a key role in cancer progression
and metastasis. Fig. 4 shows the typical degradation of
heparan sulphate by HT1080 cells stably expressing
heparanase. We recently developed an homogeneous assay
for measuring heparanase activity, which do not require any
complex equipment. Rather, it required strong human
expertises both in chemistry and biology, which is a specificity
of our platform. Due to the use of only costless equipments, it
can be easily transferred to other labs interest in the
identification of heparanase inhibitors.
Foveau B, Leroy C, Ancot F et al.
Cell Death Differentiation, 2007, 14:752-764.
Host H, Drobecq H, Locht C et al.
FEMS Microbiol Letters, 2007, 268:144-150.
Ji Z, Degerny C, Vintonenko N et al.
ubiquitinylation.
Oncogene, 2007, 26:395-406.
Lenglet G, Depauw S, Mendy D et al.
Direct recognition of the S23906-1/DNA adduct by nuclear proteins :
Implication of the glyceraldehyde-3-phosphate-dehydrogenase (GAPDH)
and HMG-B1 proteins.
Mol Cancer Therapeut, 2007, 6:3422S-S.
Marcon L, Stievenard D, Melnyk O.
Electrical detection of human immunoglobulins G from human serum
using a microbiosensor.
Biosensors & Bioelectronics, 2007, 23:81-87.
Fig. 4. Degradation of
fluorescently-labelled heparan
sulfate (HS) by HT1080 cells
stably expressing heparanase
(clones C2, C3) versus HT1080
cells wild-type (WT) or stably
expressing empty vector (-) in a
conventional assay. After
degradation, HS molecules were
separated by gel electrophoresis
and detected using a Tecan
confocal fluorescence scanner.
Medina-Palazon C, Gruffat H, Mure F et al.
Protein kinase CK2 phosphorylation of EB2 regulates its function in the
production of Epstein-Barr virus infectious viral particles.
J Virol, 2007, 81:11850-11860.
Roux M, Chai F, Ollivier N et al.
Ti-Cp functionalization by deposition of organic/inorganic silica
nanoparticles.
Biomolecular Engineering 2007; 24:549-54.
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Marcon L, Melnyk O, Stievenard D.
Current based antibodies detection from human serum enhanced by
secondary antibodies labelled with gold nanoparticles immobilized in a
nanogap.
Biosensors & Bioelectronics, 2008a, 23:1185-1188.
Imaging of protein layers with an optical microscope for the
characterization of peptide microarrays.
J Peptide Sci, 2007, 13:451-457.
Yan Y, Tulasne D, Browaeys E et al.
Int J Parasitol, 2007, 37:1539-1550.
Marcon L, Stievenard D, Melnyk O.
Characterization of nanogap chemical reactivity using peptide-capped
gold nanoparticles and electrical detection.
Bioconjug Chem, 2008b, 19:802-805.
2008
Ollivier N, Blanpain A, Besret S et al.
Synthesis of azapeptides by chemical ligation.
J Peptide Sci, 2008, 14:76.
Allan G, Barbet S, Coffinier Y et al.
Fundamental studies in nanosciences at the Institute of Electronics,
Microelectronics, and Nanotechnology (IEMN).
Int J Nanotechnol, 2008, 5:631-648.
Pinet F, Beseme O, Cieniewski-Bernard C et al.
Proteomics, 2008, 8:1798-1808.
Besret S, Ollivier N, Blanpain A et al.
Thiocarbamate-linked peptides by chemoselective peptide ligation.
J Peptide Sci, 2008a, 14:1244-1250.
Piret G, Coffinier Y, Roux C et al.
Biomolecule and nanoparticle transfer on patterned and heterogeneously wetted superhydrophobic silicon nanowire surfaces.
Langmuir, 2008, 24:1670-1672.
Besret S, Ollivier N, Blanpain A et al.
Thiocarbamate-linked peptides by chemoselective peptide ligation using
phenylthiocarbarnate chemistry.
J Peptide Sci, 2008b, 14:57.
Rocha-Perugini V, Montpellier C, Delgrange D et al.
The CD81 Partner EWI-2wint Inhibits Hepatitis C Virus Entry.
PLOS One, 2008, 3.
Cieniewski-Bernard C, Mulder P, Henry JP et al.
Proteomic Analysis of Left Ventricular Remodeling in an Experimental
Model of Heart Failure.
J Proteome Res, 2008, 7:5004-5016.
2009
Deheuninck J, Foveau B, Goormachtigh G et al.
Caspase cleavage of the MET receptor generates an HGF interfering
fragment.
Acosta-Martin AE, Chwastyniak M, Beseme O et al.
Impact of incomplete DNase I treatment on human macrophage
proteome analysis.
Proteomics Clin Applications, 2009, 3:1236-4126.
Dubois E, Cieniewski-Bernard C, Mulder P et al.
Phosphoproteome analysis of left ventricular remodeling in an
experimental model of heart failure.
Hypertension, 2008, 52:766.
Baud C, Hodak H, Willery E et al.
Mol Microbiol, 2009, 74:315-329.
Choul-li S, Drobecq H, Aumercier M.
DNA-dependent protein kinase is a novel interaction partner for Ets-1
isoforms.
Biochem Biophys Res Comm, 2009a, 390:839-844.
Dupont A, Chwastyniak M, Beseme O et al.
human macrophages.
J Proteome Res, 2008, 7:3572-3582.
Choul-li S, Laitem C, Roland I et al.
DNA-PK and PARP-1, novel interaction partners of Ets-1 transcription
factor.
FEBS J, 2009b, 276:370.
Gouyer V, Fontaine D, Dumont P et al.
Autocrine induction of invasion and metastasis by tumor-associated
trypsin inhibitor in human colon cancer cells.
Oncogene, 2008, 27:4024-4033.
Deheuninck J, Goormachtigh G, Foveau B et al.
Phosphorylation of the MET receptor on juxtamembrane tyrosine
residue 1001 inhibits its caspase-dependent cleavage.
Cell Signalling, 2009, 21:1455-1463.
Hahne JC, Okuducu AF, Sahin A et al.
The Transcription Factor ETS-1: Its Role in Tumour Development and
Strategies for its Inhibition.
Mini-Reviews Med Chem, 2008, 8:1095-1105.
Dubois E, Cieniewski-Bernard C, Mulder P et al.
Study of Post-Translational Modifications of Contractile Proteins in Left
Ventricular Remodeling.
Hypertension, 2009, 54:1185.
Hochart-Behra AC, Behra-Miellet J, Sam J et al.
Antioxidative effect of Bacteroides thetaiotaomicron extracts: superoxide
dismutase identification.
Analyt Bioanalyt Chem, 008,391:415-43.
Fertier L, Cretin M, Rolland M et al.
Love wave immunosensor for antibody recognition using an innovative
semicarbazide surface functionalization.
Sensors Actuators B-Chem, 2009a, 140:616-622.
Hodak H, Wohlkonig A, Smet-Nocca C et al.
J Mol Biol, 2008,376:414-46.
Fertier L, Rolland M, Persin M et al.
Surface Modifications of Love Acoustic Waves Sensors for Chemical and
Biological Detection.
Sensor Letters, 2009b, 7:750-756.
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Foveau B, Ancot F, Leroy C et al.
Down-Regulation of the Met Receptor Tyrosine Kinase by Presenilindependent Regulated Intramembrane Proteolysis.
Mol Biol Cell, 2009, 20:2495-2507.
Mhidia R, Melnyk O.
Selective cleavage of an azaGly peptide bond by copper(II). Long-range
effect of histidine residue.
J Pept Sci, 2010a, 16:141-147.
Helleboid-Chapman A, Nowak M, Helleboid S et al.
Apolipoprotein A-V Modulates Insulin Secretion in Pancreatic betacells Through its Interaction with Midkine.
Mhidia R, Vallin A, Ollivier N et al.
Synthesis of Peptide-protein conjugates using N-succinimidyl carbamate
chemistry.
Bioconjug Chem, 2010b, 21:219-228.
Juillard F, Hiriart E, Sergeant N et al.
Epstein-Barr Virus Protein EB2 Contains an N-Terminal Transferable
Nuclear Export Signal That Promotes Nucleocytoplasmic Export by
Directly Binding TAP/NXF1.
Virology 2009; 83:12759-68.
Piret G, Drobecq H, Coffinier Y et al.
Matrix-Free Laser Desorption/Ionization Mass Spectrometry on Silicon
Nanowire Arrays Prepared by Chemical Etching of Crystalline Silicon.
Langmuir, 2010, 26:1354-1361.
Lenglet G, Depauw S, Drobecq H et al.
Involvement of GAPDH in DNA adduct recognition : implication for a DNA
destabilizing compound.
Ejc Supplements, 2009, 7:125.
Lerouge F, Melnyk O, Durand JO et al.
Towards thrombosis-targeted zeolite nanoparticles for laser-polarized
Xe-129 MRI.
J Materials Chem, 2009, 19:379-386.
Ollivier N, Besret S, Blanpain A et al.
Silver-Catalyzed azaGly Ligation. Application to the Synthesis of
Azapeptides and of Lipid-Peptide Conjugates.
Bioconjug Chem, 2009, 20:1397-1403.
Pamelard F, Even G, Apostol C et al.
PASE: A Web-Based Platform for Peptide/Protein Microarray Experiments.
Methods Mol Biol, 2009, 570:413-430.
Sahin A, Vercamer C, Kaminski A et al.
Ets 1 target genes.
Int J Oncol, 2009, 34:377-389.
Salhi B, Vaurette F, Grandidier B et al.
The collagen assisted self-assembly of silicon nanowires.
Nanotechnology, 2009, 20.
In Situ Ligation between Peptides and Silica Nanoparticles for Making
Peptide Microarrays on Polycarbonate.
Bioconjug Chem, 2009a, 20:550-557.
Souplet V, Roux C, Melnyk O.
Peptide microarrays on bisphenol a polycarbonate.
Methods Mol Biol, 2009b, 570:287-297.
Stechly L, Morelle W, Dessein AF et al.
Galectin-4-Regulated Delivery of Glycoproteins to the Brush Border
Membrane of Enterocyte-Like Cells.
Traffic, 2009, 10:438-450.
Verreman K, Baert JL, Drobecq H et al.
CoAA (CoActivator Activator) increases the activity of the ETS-related
transcription factor ERM by modulating its sumoylation.
FEBS J, 2009, 276:394.
2010
Marcon L, Wang M, Coffinier Y et al.
Photochemical Immobilization of Proteins and Peptides on
Benzophenone-Terminated Boron-Doped Diamond Surfaces.
Langmuir, 2010, 26:1075-80.
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Macromolecular
crystallography
CNRS UMR 8161
Vincent VILLERET
Contact :
INRI Lille 1
00 33 3 62 53 17 42
Group Members :
Villeret Vincent, DR2 CNRS
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Structural Biology - platform of
Macromolecular crystallography
J Mol Biol, 2007, 370:93-106.
The mission of the macromolecular crystallography platform
is to provide to biologists of the campus and at the regional
level an access to techniques of structural biology, in particular
to X-ray diffraction. The platform is fully equipped to develop
studies from cloning of proteins of interest to their final
structural analysis. The platform comprises a molecular
biology lab, with all the necessary equipments for cloning,
expression and purification of macromolecules, crystallization
screening at various temperatures, either manually or with a
crystallization robot, mounting of crystals in capillaries or in
cryo loops. The laboratory is also fully operational for X-ray
data collection, with a rotating anode X-ray generator
equipped with confocal mirrors, a Mar345 Imaging Plate
detector, and also a cryogenic Oxford system. Storage of
frozen crystals is also available in the lab, and platform users
have a frequent access to synchrotron beamlines, in particular
to the ESRF synchrotron in Grenoble.
Verbelen C, raze D, Dewitte F, Locht C, Dufrêne YF.
interactions.
J Bacteriol, 2007, 24:8801-8806.
Wohlkönig A, Sénéchal M, Dewitte F, Backers K, Emeux C, Villeret V.
Expression and purification in high yield of a functionally active
recombinant human Type I inositol(1,4,5)P3 5-phosphatase.
Protein Expr Purif, 2007, 55:69-74.
2008
Huet J, Azarkan M, Looze Y, VilleretV, Wintjens R.
Crystallization and preliminary X-ray analysis of a family 19 glycosyl
hydrolase from Carica papaya latex.
Huet J, Rucktooa P, Clantin B, Azarkan M, Looze Y, Villeret V, Wintjens R.
X-ray Structure of Papaya Chitinase Reveals the Substrate Binding Mode
of Glycosyl Hydrolase Family 19 Chitinases.
Biochemistry, 2008, in press.
- The first research axis developed on the platform is the
structural study of mecanisms underlying bacterial
pathogenesis. In collaboration with microbiologists of the
campus, we are pursuing the study of surface structures of
pathogenic bacteria, such as adhesins, protein export
mecanisms, or mecanisms of bacterial cell wall synthesis. More
specifically, we have studied type III and type V secretion
systems. We have also characterized mechanisms of
periplasmic transport and of modulation of virulence. Finally,
the platform is involved in structural drug design programs of
new drugs directed against the cell wall synthesis in
Mycobacterium tuberculosis.
Deletion of flagellin's hypervariable region abrogates antibody-mediated
neutralization and systemic activation of TLR5-dependent immunity.
J Immunol, 2008, 181:2036-2043.
Wohlkönig A, Hodak H, Clantin B, Sénéchal M, Bompard C,
Jacob-Dubuisson F, Villeret V.
Crystallization and preliminary X-ray diffraction analysis of the PeptidylProline Isomerase Par27 of Bordetella pertussis.
- The second research axis is the study of important proteins
in humain biology. We study the SUMO modification of
humain proteins involved in various cancers. So far, limited
information is available regarding the way this important
modification acts on the protein conformations and functions.
We use the platform resources to develop the structural
approach of this post-transcriptional modification. We are also
studying the family of human inositol phosphatases, which
are crucial enzymes for signal transmission in eukaryotic cells.
2009
Clantin B, Leyrat C, Wohlkönig A, Hodak H, Ribeiro ED Jr, Martinez N,
Baud C, Smet-Nocca C, Villeret V, Jacob-Dubuisson F, Jamin M.
Structure and plasticity of the peptidyl-prolyl isomerase Par27 of Bordetella
pertussis revealed by X-ray diffraction and small-angle X-ray scattering.
J Struct Biol, 2009, Nov 20.
Villeret V, Tulasne D, Fafeur V. 2009
1001 inhibit its caspase-dependent cleavage.
Cell Signal, 2009, 21:1455-1463.
Publications
First structural insignts into the TpsB/Omp85 superfamily.
Biol Chem, 2009, 390:675-684. Review.
2007
Clantin B, Delattre AS, Rucktooa P, Saint N, Albano C, Locht C,
Jacob-Dubuisson F, Villeret V.
Structure of the membrane protein FhaC : a member of the Omp85/TpsB
Science, 2007, 317:957-961.
Leroux F, Willery E, Mathys V, Déprez-Poulain R, Delcroix G, Frénois F,
Aumercier M, Locht C, Villeret V, Déprez B, Baulard AR. 2009
Nat Med, 2009,15:537-544.
Herrou J, Bompard C, Antoine R, Leroy A, Rucktooa P, Hot D,
Huvent I, Locht C, Villeret V, Jacob-Dubuisson F.
Structure-based mechanism of ligand binding for periplasmic solutebinding protein of the Bug family.
J Mol Biol, 2007, 373:954-64.
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HTS and ADME-PK
screening lab
Inserm U761
Benoit DéPREZ
Contact :
00 33 3 20 96 40 24
[email protected]
Group members :
Déprez Benoît, PU Lille 2
Leroux Florence, IR Inserm
Host Hélène, Postodoctoral fellow Inserm
Piveteau Catherine, Postodoctoral fellow Lille 2
Dumont Julie, AI Lille 2
Dassonneville Sandrine, Technician IPL
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used to study enzymatic reactions, protein binding, proteinprotein interactions, DNA-protein interactions. Except a few
immuno-assays, most of the tests are homogeneous
screening assays, highly convenient to perform HTS. Several
cell-based assays using reporter genes were also developed
(TGR5 agonist assay, EthR inhibition assay). In total more than
20 assays were developed and screened in the past 3 years.
The main equipments are : (1) two multilabel multimodefluorescence / luminescence reader,(2) a lightcycler 480
real-time detection system (Roche), used to evaluate protein
ligand binding in thermal shift assays, (3) a pipettor device and
(3) a Genesis RSP instrument (Tecan) for serial dilutions. A cell
culture unit is also available to implement cell-based assays.
Mission of the platform
Deprez’ team (INSERM U761, “Biostructures and Drug
Discovery”) is specialized in the identification and
optimization of small organic molecules that modulate
biological processes. It is composed of experts is in the fields
of medicinal chemistry (from chemistry to pharmacokinetics),
high throughput screening and chemoinformatics who
collaborate with biologists working on novel target discovery
and validation. Their mission is to design and prepare ligands
modulating the effects of these proteins targets in vitro first
and in animal model ultimately. These compounds serve first
as pharmacological probe to validate therapeutic approaches
and then as drug prototypes. Their know-how is a
combination of academic and industrial experience. They
collaborate with academic labs (Pasteur Lille, Paris Descartes ;
U.Gent, Belgium; U.Chicago, USA; Imperial College, UK,
Pasteur Institute, Korea) and industrial researchers (Galderma,
Cytomics, Diverchim, Targeon). Deprez et al are in charge
of the regional screening platform and all the required
databases and interfaces for its use (this platform is accredited
by GIS IBISA). The laboratory is also a founder member of
PRIM, the regional consortium for drug discovery
(www.drugdiscoverylille.fr) Pr. Benoit Deprez is a recognized
expert in medicinal chemistry who has been previously
in charge of the development of screening and lead
optimization programs in two world-known biotech
companies (Cerep SA and Devgen NV). He is also expert for
several SME involved in drug discovery and Venture Capital
investors. Through active collaborations, the team is
constantly exposed to academic and industrial drug discovery
projects and their needs.
Compound profiling, beyond «on-target» activity
(Florence Leroux)
To enable the generation of relevant in vivo data, we have
developed a procedure to predict and eventually measure the
exposure of an animal model to a given compound. This
profile comprises data on physical chemical properties known
to govern bioavailability (logD7.4, aqueous solubility,
microsomal stability…). Only compounds displaying the
desired ADME properties, beyond their activity on target are
administered to animal. This saves animal lives and
researcher’s time. Importantly the knowledge generated by
these studies is applicable to virtually any chemical series,
independently of the mode of action. Thus all the data are
stored safely in a database that can be interrogated by any
researcher in the lab and serve as learning sets for molecular
modelling. In collaboration with Juergen Siepmann, professor
of pharmaceutical sciences in Lille, we have obtained a grant
from Région Nord-Pas-de-Calais, to set up a small platform for
early formulation & early pharmacokinetics. By e-formulation
we mean rapid and miniaturized formulation, adapted to
molecules not yet fully optimized and to an administration
of minute quantities to small animals (mice in particular).
Sample preparation and bioanalysis are performed using our
LC-MSMS platform.
In September this year, a consortium of 5 labs lead by Benoit
Déprez has applied to the Equipex call for project, to set up a
world class formulation ADME platform dedicated to early
preclinical phases of drug discovery. The aim of the project,
called PharmaR3 is to combine advanced mass spectrometry,
mass spectrometry imaging and miniaturized formulation
tools to accelerate target validation in vivo, and refinereduce-replace animal testing.
Description of the platform
Chemical library and screening (Florence Leroux)
During the past 4 years, our lab has assembled a library of
65.000 compounds (“no string attached”) formatted for
screening in 96-well plates. Compounds have been selected in
collaboration with the team of Bruno Villoutreix (Inserm MTi,
Paris) from commercial vendors or prepared by our chemists
using state-of-the-art selection and design criteria, in terms of
diversity and “drug/lead-likeness”. Our sample management
system avoids repeated freeze thaw cycles and ensures the
longest possible lifetime for all the samples. Sample tracking is
seamless throughout the sample’s life (from ordering to testing).
To manage compounds and associated results, a highthroughput data analysis and mining system has been
implemented, using the Pipeline ¨Pilot™ software.
Compound screening, hit-to-lead and lead optimisation
The laboratory has a recognized expertise in the development
of miniaturized, fast and robust assays for medium to high
throughput screening. All critical screening parameters are
optimized in terms of reagent cost, required manpower and
time, as well as its discriminating power, as measured by the
Z’ and Z factors. Fluorescence data (FRET, HTRF, fluorescence
anisotropy, fluorescence intensity) or absorbance data are
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Nuclear Magnetic
Resonance
CNRS UMR 8576
Guy LIPPENS
Contact :
00 33 3 20 33 72 41
00 33 3 20 87 73 07
mail : [email protected]
Group members :
Lippens Guy, DR1 CNRS
Fritzinger Bernd, IR Région
Landrieu Isabelle, CR1 CNRS
Hanoulle Xavier, CR2 CNRS
Sibille Nathalie, CR2 CNRS
Leroy Arnaud, MCU Univ Paris XI
Smet Caroline, MCU Lille 1
Trivelli Xavier, IR2 Lille 1
Wieruszeski Jean-Michel, IR1 CNRS
Bonachera Fanny, IE CNRS
Amniai Laziza, PhD Student Lille 1
Daccache Anthony, PhD Student Lille 1
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interpretation of the biomolecular spectra is itself quite
specialized. The majority of the projects hence involve a
collaboration with the interested biochemists. After defining
the problem in terms of spectroscopy and an agreement on
sample preparation (that often involves the incorporation of
stable isotopes in the biomolecule), the facility will record the
NMR spectra and interpret them in order to obtain the
tridimensional structure of the biomolécule.
The NMR facility of the Campus equally has access to the
800MHz spectrometer at the University of Lille I, and also to
the recently installed 900MHz installed in November 2009 at
the Haute Borne Campus in Villeneuve d’Ascq. The former
spectrometer is equipped for a dual use in liquid and solid
state NMR, and has two different High Resolution Magic Angle
Spinning probe heads. It is therefore particularly well suited
for the study of heterogeneous samples such as intact
bacteria. The latter has a cryogenic probe head installed, and
thus gives the supreme resolution and sensitivity.
1) Mission of the facility
Give acces to Nuclear Magnetic Resonance for the analysis of
molecules coming from organic synthesis, and for
biomolecules.
2) Description of the facility
The facility is composed of two spectrometers, one at 300MHz
and one at 600MHz. Small organic molecules coming from the
organic chemistry groups from the campus are analysed with
the 300MHz. The operator, Mme E. Boll (Technicienne CNRS) will
acquire the spectra, process them and give a paper trace from
the resulting spectra The 600MHz spectrometer is mainly used
for applications in the field of structural and analytical
biochemistry and biology. Equiped with a cryogenic probe
head, whereby noise in the probe head is limited to obtain a
superior sensitivity, the spectrometer can analyse biomolecules
at concentrations down to the low micromolar range. For the
600MHz, with Dr. J.-M. Wieruszeski (IR1 CNRS) as person in
charge, not only the data acquisition but equally the
3) Projects
The 300MHz spectrometer performs mainly the analysis of
small organic molecules coming from the chemistry groups
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on campus. The field of study of the 600MHz spectrometer
ranges from the structural analysis of proteins in solution, the
identification of complex sugar motifs, the elucidation of posttranscriptional modifications such as phosphorylation, … The
detection and quantification of biomolecular interactions is a
well established and important application of the NMR facility.
Projets in collaboration with other groups of the campus
Calmette involve the study of the natively unfolded tail of the
Mycobacterial heparin binding hemaglutinin, with its possible
post-translational modifications (PTMs) (collaboration with Dr
D Raze and Dr C Locht), the study of the inhibitor of the PP1
phosphatase of Plasmodium falciparum (collaboration with
Dr J Khalife) and a novel prolyl cis/trans isomerase from B
pertussis (collaboration with Dr F Jacob-Dubuisson). He group
itself is centered on the neuronal protein Tau and its PTMs, and
equally on the study of the natively unfolded NS5A protein of
Hepatitis C.
2010
B, Wieruszeski JM, Lippens G.
Spectroscopic studies of GSK3{beta} phosphorylation of the neuronal tau
protein and its interaction with the N-terminal domain of apolipoprotein
E. Leroy A, Landrieu I, Huvent I, Legrand D, Codeville
J Biol Chem. 2010 Aug 2. [Epub ahead of print]
Landrieu I, Leroy A, Smet-Nocca C, Huvent I, Amniai L, Hamdane M,
Sibille N, Buée L, Wieruszeski JM, Lippens G.
2.NMR spectroscopy of the neuronal tau protein : normal function and
implication in Alzheimer's disease.
Biochem Soc Trans. 2010 Aug;38(4):1006-11.
Landrieu I, Hanoulle X, Bonachera F, Hamel A, Sibille N, Yin Y,
Wieruszeski JM, Horvath D, Wei Q, Vuagniaux G, Lippens G.
Structural basis for the non-immunosuppressive character of the
cyclosporin A analogue Debio 025.
Biochemistry. 2010 Jun 8;49(22):4679-86.
Publications
2007
Sillen A, Barbier P, Landrieu I, Lefebvre S, Wieruszeski JM, Leroy A,
Peyrot V, Lippens G.
NMR investigation of the interaction between the neuronal protein tau
and the microtubules.
Biochemistry, 2007, 46:3055-64.
2008
Bodart JF, Wieruszeski JM, Amniai L, Leroy A, Landrieu I, RousseauLescuyer A, Vilain JP, Lippens G.
NMR observation of Tau in Xenopus oocytes.
J Magn Reson, 2008, 192:252-257.
Hodak H, Wohlkönig A, Smet-Nocca C, Drobecq H, Wieruszeski JM,
Sénéchal M, Landrieu I, Locht C, Jamin M, Jacob-Dubuisson F.
J Mol Biol, 2008, 376:414-426.
Rodius S, Chaloin O, Moes M, Schaffner-Reckinger E, Landrieu I,
Lippens G, Lin M, Zhang J, Kieffer N.
The Talin Rod IBS2 {alpha}-Helix Interacts with the {beta}3 Integrin
Cytoplasmic Tail Membrane-proximal Helix by Establishing Charge
Complementary Salt Bridges.
J Biol Chem, 2008, 283:24212-24223.
Wieruszeski JM, Fritzinger B, Hanoulle X, Martins JC, Lippens G.
Sandwich-ELISE NMR : Reducing the sample volume of NMR samples.
J Magn Reson, 2008, 193:37-40.
2009
Amniai L, Barbier P, Sillen A, Wieruszeski JM, Peyrot V, Lippens G,
Landrieu I.
Alzheimer disease specific phosphoepitopes of Tau interfere with
assembly of tubulin but not binding to microtubules. FASEB J, 2009,
23:1146-1152.
Hanoulle X, Badillo A, Wieruszeski JM, Verdegem D, Landrieu I,
Bartenschlager R, Penin F, Lippens G.
Hepatitis C virus NS5A protein is a substrate for the peptidyl-prolyl
cis/trans isomerase activity of cyclophilins A and B.
J Biol Chem. 2009 May 15;284(20):13589-13601.
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Animal Unit
Jean-Pierre DECAVEL
Contact :
00 33 3 20 87 79 50
[email protected]
Group members :
Decavel Jean-Pierre, Manager DR2 IPL
Hannebique David, AI TCN CNRS
Chassat Thierry, ITA Assistant IPL
Couvreur Bruno, ITA TR2 IPL
Deudon Jean-Claude, ITA AG3 IPL
Fleurbaix Emile, ITA TCS Inserm
Lepage Francis, ITA AJT-PR Inserm
Messiaen Nathalie, ITA TCN Inserm
Mouray Anthony, ITA TR3 IPL
Persoons Philippe, ITA TR4 IPL
Riva Philippe, ITA AG3 IPL
Rogeaux Marie-Christine, ITA AGT3 IPL
Vangasbecq Marc, ITA TR1 IPL
Waroquier Gilles, ITA TR4 IPL
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The Animal unit of the Institut Pasteur de lille is a facility
meeting the needs for animal experimentation for all research
groups and for expertises and their associated organizations,
placing at their disposal :
e/ Insulators quarantine zone equipped with 8 insulators (30
m2) allowing to isolate animals of unknown or doubtful
medical status.
f/ Embryo-transfer technology zone 45 m2 surfaces equipped
with 4 insulators type of barrier : S.O.P.F.
• Buildings, equipment and facilities in conformity and
approved by the official standards and regulations (French,
European, international), fitting the activities and the
scientific requirements.
g/ Health Monitoring Laboratory 12 m2 surfaces (rat - mouse).
h/ Cryo-embryo and sperm preservation Laboratory
22 m2 surface.
Materials : 1 drying oven CO2, 1 minicool 40pc, 1MVE xlc
500 for storage in nitrogen.
• Qualified worker are involved in the care and the
monitoring of the animals in experimentation and their
environment, but also in the development of high animal
technology such as the cryo-embryo preservation or cryosperm technology, the embryos transfer decontamination
technology or aseptic hysterectomy, the exchange and the
reactivation of cryo-embryos, medical controls and
trainings within the field of animal experimentation
i/ Conventional Zone 490 m2 Surface for the housing of
conventional animals and of different species (mouse, rat,
hamster, guinea-pig,…).
j/ A2 containment, equipped with IVC in depression
(22 m2) allowing to isolate the animals in containment A2.
k/ A3 containment in the hight security laboratory NSB 3-4,
equipped with 6 insulators allowing to isolate from the
animals in containment A3.
B - Average data processing
• Registering of the animals on computer “4D”.
• Centralized technical management.
• Access control zones on computerized power station.
A - Building, facilities
and equipment m2 acres
These facilites encompass a total surface of 1.601 m2, and are
distributed on two sites and 11 zones inside the campus from
the Institut Pasteur of Lille.
a/ S.P.F. zone (Specific Pathogen Free)
420 m2 of surface for housing SPF animals.
b/ Isolators zone “Interreg” (Specific Pathogen and
Opportunist Free : S.P.O.F.).
100 m2 of surface for S.O.P.F. animals of the transgenic mice
breeding equipped with 11 isolators
c/ Breeding insulators zone (Specific Pathogen Opportunist
Free : S.P.O.F.).
78 m2 of surface equipped with 5 hemi-diving-suit macroinsulators.
d/ Breeding and experiment insulators zone (Specific
Pathogen Opportunist Free : S.P.O.F.) equipped with 23
insulators 95 m2 surfaces.
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High Security Laboratory
Jean-Pierre DECAVEL
Contact :
00 33 3 20 87 79 50
[email protected]
Group members :
Decavel Jean-Pierre, Manager DR2 IPL
Legrand Damien, ITA TR4 IPL
Vandenabeele Nicolas, ITA TR2 IPL
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production of parasites, including the emergent disease such
as SARS or H5N1 strains.
Safety and security, versatility and evolutionarity thus guided
the design of this laboratory for which equipment was or is still
specifically designed.
The laboratory, installed on three floors :
- A facility which supports the ert treatment. This one is
sterilized by filtration, inlet and outlet, on a high efficiency
filter ULPA.
- 9 working areas or independent laboratories located in the
ground floor. These cells (laboratories) can host 25
researchers and technicians simultaneously, equipped with
14 biological safety cabinet BSC II and 6 insulators for
animals housing .
- 1 Hight Security Zone (level 4) equipped with 1 Biologcal
safety cabinet BSC III and 3 High containment insulators
(Gloves boxes),
- 1 zone equipped with three -80° freezers,
The LHS waste are decontaminated by one effluent
decontamination system and one autoclave.
The study of the emergent diseases such as SARS, bird flu, or of
diseases in outbreak like tuberculosis, require conditions of
safety reinforced both for the worker and the environment.
The High Security Laboratory L.H.S (NSB 3-4) of the Institut
Pasteur de Lille has the role to fulfill the legal requirements and
techniques of handling the GMO’s or pathogenic dangerous for
the environment (up to level 4 in BCS 3 and Gloves box) or the
public health (except the hemoragie fevers (level 4)).
The handling of the man or animal diseases always required
specific working conditions.
The concept of containment physical of the laboratories for the
handling of the micro-organisms of levels 1, 2, 3 and 4 appeared
within the U.S.A in the Seventies. This concept was created to
protect the environment during handling and the study from
the GMO's research activities or analyses in microbiology. The
study of the emergent diseases such as SARS, bird flu (H5N1),
highly pathogenic, or of the diseases such as tuberculosis, but
also the handling of GMO's request an optimal safety and
security level both for the protection of the worker and of the
environment.
Work on biological agent in the LHS is carried out with respect
to the French and European legislations , but also of the
reference on the GMO' S, or those of dangerous pathogenic
agents for the environment and the public health, it should be
noted that the French regulation is rather recent (first texts were
adopted in the years 1990) and rises primarily from the
transposition of European directives.
The decree of July 18th 1994 modified in 1997 and 1998 fixes
the list of the pathogenic biological agents. The classification
mentioned in these lists is delicate exploitation because the
associated biological risk is very lovoad and is evolving
progressively with new knowledge. It is then necessary to
consult lists coming from foreign countries, and thus with the
assistance of professional people to establish risk assessment
based on the various criteria of classifications. : severity of the
disease, dissemination in the environment; existence of
preventive or curative treatments.
The decree of July 16th, 2007 prevention, in particular of
containment, where the workers are likely to be exposed to
pathogenic biological agents.
Lastly, the law 92-654 of July 13th 1992 and the texts of its
application refer to the control of the use and the dissemination
of the GMO’S.
A good training, completed by a high level of knowledge are
requested from the personnel working the LHS.
Team
The laboratory of high safety is placed under the responsibility
of Jean-Pierre De CAVEL assisted by several technicians.
JP DE CAVEL (Other Field )
(a) French Expert in 6 th Framework Programme on Research,
Technological Development and Demostration Coodination Action:
BIOSAFETY- EUROPE.
(b) Expert for The Bureau d’Expérimentation Animale de l’INSERM
(c ) Biosasety and Biosecurity Expert for AFSSAPS
Buildings,facilities and equipment of the laboratory of high
safety are devoted to the teams of the Calmette Site, (Institut
Pasteur de lille and Institut de Biologie de Lille) including the
university or the CHRU.
The units carried out in cast solid polyester panels guarantee
an absolute level of sealing. The insulation of the cells
(laboratories) allows the reception of at least eight to ten
different research themes, getting for this structure an original
character compared to other laboratories of this type designed
to fulfill the requirements for only one pathogen.
The laboratory allows the implementation of all the techniques,
energy of the cellular culture to animal handling, the viral
Publications
Guide de l’ultra-propreté 2008 ( Technical File )
1 - Laboratoires et animaleries de sécurité biologique.
De Cavel JP.
2 - Les laboratoires de sécurité.
De Cavel JP, Morand T.
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Genomic Analysis
Laboratory
Inserm U 744
Philippe AMOUYEL
and Nathalie FIEVET-VERRECAS
Contact :
Tél. 00 33 3 20 87 71 15
Fax 00 33 3 20 87 78 94
[email protected]
Group members :
Amouyel Philippe, PU PH Lille 2
Fiévet-Verrecas Nathalie, IE IPL
Ledoux Patricia, Technician IPL
Mallet Kevin, Vacation Inserm (CDD)
Camerlynck Alisson, IPL (CDD)
Auvray Cécile, IPL (CDD)
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Nathalie FIEVET-VERRECAS, an IPL engineer, is the technical
director and is assisted by a small permanent staff of
technicians.
1) Mission
The rapid development of studies and research in the life
sciences as well as the explosion of tools for genomics and
post-genomics, beyond work in in vitro or animal models,
requires access to biological material of human origin.
For reasons essential to the protection of humans, bioethics
laws very rigorously oversee the conditions of collection,
storage and use of this human biological material. Thus any
clinical, epidemiologic or experimental study in this domain
must comply with a number of rules aimed at ensuring these
conditions are met.
3. CRB operating procedures
• Electronic control of personnel access to the site.
• Establishment of standardized procedures for quality
assurance, safety and sample traceability.
• Computerized sample management: traceability of
subjects, samples and their derivatives, biological
annotations, stock management, entries and exits, transfers
and cloning, audit of connections and data, multicriteria
research, printing barcode labels, data exportation
• Real-time control of temperatures in the various freezers : 80°C, -30°C, +4°C
• Confidentiality of information.
• Access to samples reserved exclusively to the department
that collected it.
The Genomic Analysis Laboratory, recognized as a biological
resource center (CRB) in the framework of INSERM-FRT
requests for proposals, provides teams of clinicians and
researchers with the analysis and logistic support necessary
to comply with these rules:
• homogeneity of sampling methods
• homogeneity of methods for the transformation (DNA
extraction, etc.) and processing of biological materials
• storage and preservation of samples (plasma, serum,
nucleic acids, etc.) in their initial condition, for a set or
unlimited duration, depending on the specific scientific
objectives
• information management making it possible to know at any
moment the sampling method, sample source, date, place
of storage, and all activities related to it, to ensure complete
traceability
• specific and reliable system for sample identification.
Standard procedure for management of a blood sample
The person responsible for taking the sample completes a
sample tracking document.
This anonymous form contains all information relative to the
blood sample, the treatment of the blood tubes
(centrifugation, washing…) and data related to the derived
samples obtained after treatment.
Some samples are then transferred to a laboratory where the
biochemical assays are performed; other derivatives are stored
at -80°C and constitute the biobank, while still others undergo
DNA extraction.
To enable the exchange of biological resources and the data
related to them between different centers, INSERM and the
Ministry of Research established quality assurance procedures
for CRBs in 2006. National quality assurance standards for CRBs
are now being drafted under the aegis of AFNOR.
This step is an essential prerequisite for the emergence of a
national network of biobanks.
All data from the sample tracking document are then fully
entered into the database with Databiotec sample
management software (entry of sample source, derived
products, characterization, number…)
2. Resources
4. Projects underway
2.1. Premises and equipment
The Genomic Analysis Laboratory occupies an area of 135 m2
and has a 37m2 air-conditioned room for –80°C freezers.
Its equipment includes a Microbiological Safety Cabinet, the
material and supplies necessary for work in molecular biology,
and the infrastructure necessary for sample storage, that is, 13
-80°C freezers and a system of computerized sample
management.
2.2. Personnel
The Genomic Analysis Laboratory is supervised by Philippe
AMOUYEL, University professor and hospital physician at the
University of Lille 2, Director of the UMR 744 Unit “Public
Health and Molecular Epidemiology of aging related diseases”
, Director of the Institut Pasteur de Lille and Director of the
Scientific Cooperation Foundation for Alzheimer’s disease and
related disorders
The LAG-CRB of the Institute Pasteur of Lille manage 9 national
and international collections.
The Three Cities (3-C) Study : Bordeaux, Dijon and
Montpellier.
Launched with the support of the Foundation for Medical
Research in 1997, its objective is "to estimate the risk of
dementia and severe degeneration attributable to risk factors
and to vascular disease."
The cohort includes 9692 persons older than 65 years,
followed for 4 years. Each participant undergoes a battery of
tests : questionnaire, cognitive tests, blood sample,
electrocardiogram and carotid ultrasound.
8860 subjects agreed to give blood samples ; these samples
generated 221 500 derived samples (plasma, red blood cells,
and serum), 180 550 of which are stored at LAG.
3 500 subjects had an MRI (magnetic resonance imaging).
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These images will be analyzed by automatic software that can
detect cerebral lesions and calculate tissue volume, in
particular, of gray matter.
In the long run, the 3C study should provide answers to two
essential questions : who are the people at highest risk for
whom preventive action is possible ? And to what extent does
this prevention favorably modify the disease course ?
adolescence and the development of nontransmissible diseases.
LAG extracted DNA samples for 1 143 subjects.
Cadisp : Cervical Artery Dissections and Ischemic Stroke
Patients - for France : Amiens - Dijon - Besançon - Paris (x2)
- Lille
This study, initiated in 2004, is a European Consortium
performing research on ischemic stroke in the young and in
particular on cervical artery dissection (the most common
cause of ischemic stroke stroke in young people).
The CADISP-genetic study (2006-2008) is one among other
collaborative research projects of the CADISP Consortium with
the following goals :
• Look for genetic variants associated with CAD
• Look for environmental risk factors of CAD
• Address therapeutic aspects in the setting of a multicenter
registry
LAG extracted DNA samples for 549 subjects.
Covadis : Cognitive vascular diseases - Dijon
Continuation of the 3-C Study in Dijon, this new study will make
available a larger number of vascular events and cases of
dementia by extending follow-up for 4 years in a subcohort of
subjects who had an MRI as part of the 3C study (1606 subjects
and 35 015 samples of plasma and serum) stored at LAG.
Paradigme : Parkinson and Alzheimer diseases: impact of
gene, metabolism and environment - Lille CHRU
Conducted as part of an INSERM 2002 funding program
(collection of human biological material for research
purposes), the aim of this study is to identify new factors of
genetic susceptibility involved principally in Alzheimer and
Parkinson diseases, both neurodegenerative, as well as their
potential interactions with environmental factors.
796 subjects with disease were seen between May 2003 and
January 2006. Blood samples made it possible to collect a
biobank of 18 490 derived samples (plasma, red blood cells,
and serum) stored at LAG.
Young Heart Study : Northern Ireland
Northern Ireland is one of the countries with the highest rates
of coronary disease in the world (from the age of 15 years, 1
Irish child in 4 is at risk of a heart attack). The aim of this study
is to discover the risk factors leading to the development of
these diseases in adulthood.
This study, funded by the Northern Ireland Fund, the Heart
and Stroke Association, the British Heart Foundation and the
Wellcome Trust, began in 1988.
The cohort includes 509 children aged 12 years and 506
children aged 15 years at inclusion. These children were
reexamined in the same conditions (same physical tests) in
1992 and then in 1997.
LAG extracted DNA for 444 subjects.
Euroaspire III : European Activity on Secondary and Primary
Prevention in order to Reduce Events 22 countries : Italy, France,
Belgium, Greece, the Netherlands, Slovenia, Romania, Bulgaria ,
Ireland, Lithuania, Latvia, Germany, Finland, Cyprus, Hungary,
Poland, Croatia, Spain, Czech Republic, Turkey and England.
The aim of this multicenter European study is to identify risk
factors in patients with coronary diseases.
The Lille team selected 342 subjects hospitalized for coronary
diseases. LAG is storing the 3382 French samples (plasma, sera,
white blood cell).
SU.VI.MAX : Antioxidant Vitamin and Mineral
Supplementation. France
Launched in October 1994, by the Conservatoire National des
Arts et Métiers, INSERM, the Ministries of Health, Agriculture and
Research, and the city of Paris, this study has 2 objectives :
• assess the efficacy of antioxidant vitamin and mineral
supplementation in the prevention of cardiovascular
diseases and cancer,
• construct a database of the food intake of the French
population.
Monalisa and MonalisaNut : Monitoring National Arterial
Risk - Lille, Strasbourg and Toulouse
This survey, which began in October 2005, is intended to
estimate the prevalence of cardiovascular risk factors
(hypertension, dyslipidemia, diabetes, overweight, obesity,
sedentary lifestyle, and smoking) in the population aged 3574 years in the regions of Lille, Strasbourg and Toulouse (1600
subjects randomly selected in each geographic zone).
LAG stores the samples from all 3 centers, that is to say 58 173
samples (plasma, sera) derived from the blood samples of 4
797 subjects.
LAG extracted DNA from Lille and Strasbourg samples for 3
168 subjects.
Between 1994 and 2002, 13 017 volunteers (7886 women
aged 35-60 years and 5141 men aged 45-60 years)
participated in this study.
The volunteers were allocated to one of two groups: one
received the SUVIMAX pill, and the other a placebo capsule
with no active substance, daily for 8 years. The SUVIMAX
cocktail included antioxidant nutritional supplements
composed of : Beta-carotene 6 mg, Vitamin C 120 mg, Vitamin
E 30 mg, selenium 100 µg, and zinc 20 mg.
Helena : Healthy Lifestyle in Europe by Nutrition in
Adolescence
11 countries : Greece, England, Germany, Belgium, Crete,
France, Hungary, Italy, Sweden, Austria and Spain
This study will make it possible to obtain valid and reliable
information on the nutritional status of European adolescents
(dietary habits, nutrition knowledge and eating attitudes, food
choices and preferences, metabolic profile, body composition,
vitamin status, physical activity, genetic variations, etc.) to
improve our understanding of the relations between
After eight years of follow-up (questionnaires, health
examinations, dietary surveys, plasma assays, and specific
tests in the case of diseases), the results show that this intake
of antioxidant vitamins and minerals reduced by 31% cancer
incidence and overall mortality in men. This reduction affected
most cancer sites: gastrointestinal, ORL, respiratory, cutaneous
LAG extracted DNA from samples for 955 subjects, collected
in 1994 and 1995.
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Publications
2008
Le Fur I, Laumet G, Richard F, Fievet N, Berr C, Rouaud O,
Delcourt C, Amouyel P, Lambert JC.
Association study of the CFH Y402H polymorphism with Alzheimer's
disease.
Neurobiol Aging, 2008, apr 21.
2009
Association of plasma amyloid beta with risk of dementia: the prospective
Three-City Study.
Neurology, 2009, 73:847-853.
Lambert JC, Health S, Even, Campion D, Sleegers K, Hiltunen M,
Combarros, Zelenika D, Bullido MJ, Tavernier B, Lentenneur L,
Bettens K, Berr C, Pasquier F, Fievet N, et al.
Nat Genet, 2009, 41:1047-1048.
Hauw JJ, et al.
Transcriptomic and genetic studies identify IL-33 as a candidate gene for
Mol Psychiatry, 2009 Feb 10 [Epub ahead of print].
2010
Berr C, Dartigues JF, Tzourio C, Campion D, Lathrop M, Amouyel P.
Implication of the Immune System in Alzheimer's Disease : Evidence from
Genome-Wide Pathway Analysis.
J Alzheimers Dis, 2010, Apr 22.
Laumet G, Chouraki V, Boley BG, Legry V, Heath S, Zelenika D,
Epelbaum J, Berri C, Dartigues JF, Tzourio C, Campion D, Lathrop M,
Systematic Analysis of Candidate Genes for Alzheimer's Disease in a
French, Genome-Wide Association Study.
J Alzheimers Dis, 2010, Apr 22.
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Contents
High Throughput
Genomic Platform
Genoscreen (1)
André TORDEUX
Contact :
Tél. 00 33 3 20 87 71 53
Fax 00 33 3 20 87 72 64
mail : [email protected]
website : www.genoscreen.com
Group members :
Tordeux André, CEO
Huyghe-Banse Catherine, Direction assistant
Ferreira Stéphanie, PhD – Laboratory and R&D manager
Hubans-Pierlot Christine, PhD – Bioinformatics manager
Allix-Beguec Caroline, PhD – Research project manager
Poulain Maïté, Sequencing service manager
Honvault Sophie, Sales manager
Desselle Mathilde, Marketing & communication manager
(1) Genoscreen
Prize winner 2006 - INPI Trophy - Research Category
Approved C I R (by the French Minister of Research)
223
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• Genotyping of SNPs markers by the Sequenom® technology
(recommended for the analysis of 10 to 47 SNPs)
• Genotyping of SNPs markers by the Veracode® technology
(recommended for the analysis of 48 to 1* or 3* 384 SNPs)
1) Mission of the platform
Genoscreen develops and performs innovative analytic
services in genomics on all kind of genomes (human, animal,
plants, micro-organisms), to answer the needs of research
teams in the fields of Health, Environment, Agriculture,
AgroFood, …
iii. Molecular microbiology
• Microorganisms typing by the MLST method (bacterias and
other microorganisms)
• Staphylococcus aureus typing by the SPA method
• Mycobacterium tuberculosis typing by the MIRU method (9,
12, 15 or 24 MIRU markers)
• MIRU-VNTR Typing Kit - Mycobacterium tuberculosis complex
24 loci – 100 tests
• MIRU-VNTR Calibration Kit - Mycobacterium tuberculosis
complex
• MIRU-VNTR Validation Kit - Mycobacterium tuberculosis
complex
There are three types of the platform activities :
a/ Standard and custom analytical services, under quality
process, on any type of genome (human, animal, plant
and micro-organisms)
b/ Support for research teams through scientific
partnerships
c/ Training in genetic analyses using the most recent
techniques
Bioinformatics
a/ Proposed services :
•
•
•
•
•
•
•
•
•
•
•
DNA sequencing
• DNA sequencing by Sanger technology (3730XL®, Applied
Biosystems®)
• De novo sequencing and re-sequencing by ultra high
throughput pyrosequencing (GS FLX®, Roche Diagnostics®)
• Mutation detection by Sanger sequencing (3730XL®,
Applied Biosystems®)
• Mutation detection by ultra high throughput
pyrosequencing (GS FLX®, Roche Diagnostics®)
• Microorganisms identification by constitutive genes
sequencing ex : 16S (3730XL®, Applied Biosystems®) and
bioinformatics analysis
• 16 S metagenomics by ultra high throughput pyrosequencing (GS FLX®, Roche Diagnostics®)
• De novo metagenomics by ultra high throughput
• Ultra high throughput transcriptomic sequencing (GS FLX®,
Roche Diagnostics®)
Specific primer design
Phylogenetic analysis
Mutation detection
Metagenomes analysis
Genomes assembling and annotation
Transcriptome analysis
Genomes comparison and analysis
Protein comparison and analysis
Statistic analysis of allelic variants
MIRU-VNTR Data Manager Software
On-site trainings
Other genomic services
• Genetic materials extraction (gDNA, total RNA, plasmids,
BAC etc.) from various kinds of samples (blood, plants,
tissues, bacterias etc)
• Gene expression by TaqMan® or Veracode® technology
• Cloning and screening by sequencing (3730XL®, Applied
Biosystems®)
• MIRU-VNTR typing training
• Tailored analysis
Genotyping
b/ Examples of scientific partnerships :
i. Microsatellites markers
• Production of microsatellites markers by cloning and
capillary sequencing (3730XL®, Applied Biosystems®)
• Production of microsatellites markers by high throughput
• Development of microsatellites markers by fragment
analysis on agarose gel
• Development of microsatellites markers by fragment
analysis on capillary sequencer (3730XL®, Applied
Biosystems®)
• Genotyping of microsatellites markers by fragment analysis
on capillary sequencer (3730XL®, Applied Biosystems®)
The aim of the platform is to solve research problems,
eventually, in collaboration with partners, to put in place a
research program. Those partnerships are developed through
research agreements.
In this frame, the platform naturally finds its place beside
teams which answer to calls of research projects (A N R
[National Agency for Research], VIIth PCRD...).
c/ The training offer :
In response to its mission to facilitate research in life sciences,
Genoscreen offers various activities dedicated to the
information and the training of students, researchers and
Heads of research teams.
ii. SNPs markers
• Genotyping of SNPs markers by the TaqMan® technology
(recommended for the analysis of less than 10 SNPs)
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- Professional qualification and student trainings organized
in-house or on-site (bioinformatics, MIRU-VNTR typing,
gene expression,…) - Training provider registered under the
n° 31 59 06657 59 (DRTEFP Lille, France).
- Inter or intra-laboratories specific conferences related to the
services offered and the developed technologies
- Visits of the platform with presentation of the materials and
explanation of the technologies implemented
Publications
One of the interests of the platform is that its contribution is
limited (for the majority of the projects) to a technical
participation without any claim to neither reference name on
the publications nor to participation in any intellectual
property.
However, the scientific services provided by the platform can
lead its staff to be associated to the publication authors.
Examples :
Chapuis J, Hot D, Hansmanel F, Kerdraon O, Ferreira S, Hubans C,
Maurage CA, Huot L,Bensemain F, Laumet G, Ayral AM, Hauw JJ,
DeKosky ST, Lemoine Y, Iwatsubo T,Wavrant-Devrièze F, Buée L,
Pasquier F, Berr C, Mann D, Lendon C, Kamboh MI, Amouyel P,
Lambert JC.
Transcriptomic and genetic studies identify IL33 as a candidate gene for
Mol Psychiatry, 2009 Jan.
Shamputa IC, Lee J, Allix-Béguec C, Cho EJ, Lee JI, Rajan V, Lee EG,
Min JH, Carroll MW, Goldfeder LC, Kim JH, Kang HS, Hwang S,
Eum SY, Park SK, Lee H, Supply P, Cho SN, Via LE, Barry CE 3rd.
Genetic diversity of Mycobacterium tuberculosis isolates from a tertiary
tuberculosis hospital in South Korea.
J Clin Microbiol, 2009 Dec.
2) Description of the platform
A broad range of techniques offering optimized analyses.
Moreover, on initiative of publications’ authors, the platform
services are generally mentioned in the “Materials and
methods” chapter of the publications they lead.
• PCR platform, Applied Biosystems®
• Sequencers: 3730 XL (Applied Biosystems®) automatic
capillary sequencer, GS-FLX 454 (Roche Diagnostics ®)
• Genotyping technologies: Taqman® SNPs Genotyping Assay
on 7900 HT (Applied Biosystems®), Veracode® technology
on BeadXpress® (Illumina®)
• Bacterial typing by techniques of reference and advanced
innovating techniques (MLST, MLVA, SPA-typing, MIRUVNTR,….)
• qPCR, Transgenomic® dHPLC , Tecan® robot,…
Examples :
Mallard K, Sharaf Eldin GS, McNerney R.
ScreenTape as a tool for the rapid differentiation of Mycobacterium
tuberculosis isolates.
J Med Microbiol, 2009 Sept.
Lesobre L, Lacroix F, Caizergues A, Hingrat Y, Chalah T,
Saint Jalme M.
Conservation genetics of Houbara Bustard (Chlamydotis undulate
undulata): population structure and its implications for the reinforcement
of wild populations.
Conservation Genetics, 2009 Sept.
3) Staff
A team of high level professionals in interaction with many
scientific teams throughout the world.
20 people work full-time in the platform among them doctors,
engineers and technicians. As well as this strong permanent
manpower, the platform also profit from the expertise of
senior scientific consultants.
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Contact - Institut Pasteur

Transcription

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