Report 2009 - Turku Centre for Biotechnology
Transcription
Report 2009 - Turku Centre for Biotechnology
TURUN BIOTEKNIIKAN KESKUS ÅBO BIOTEKNIKCENTRUM TURKU CENTRE FOR BIOTECHNOLOGY th 0 2 y p y p r a a s H nniver A Cell Signalling to Systems Biology Research TURKU CENTRE FOR BIOTECHNOLOGY REPORT 2009 TURUN BIOTEKNIIKAN KESKUS Tykistökatu 6 B P.O.BOX 123 FI 20521 Turku, Finland Tel: +358 2 333 8603, Fax 358 2 333 8000 Annual Report 2009 Turku Centre for Biotechnology Published by: Turku Centre for Biotechnology P.O. Box 123, FI-20521 Turku, Finland Tel. int. +358-2-333 8603, fax int. +358-2-333 8000 http://www.btk.fi Editorial Board Riitta Lahesmaa (Chair) Tero Aittokallio Eleanor Coffey Garry Corthals Michael Courtney Konstantin Denessiouk Attila Gyenesei John Eriksson Jyrki Heino Johanna Ivaska Panu Jaakkola Marko Kallio Olli Kallioniemi Päivi Koskinen Linnéa Linko Tassos Papageorgiou Lea Sistonen Jukka Westermarck Photographs: KUV@TEHDAS Roni Lehti, Photograph archives of the Centre for Biotechnology. Front cover image: Patrik Jones, back cover images: (2nd from top) Tassos Papageorgiou, (bottom) Marko Kallio, Photograph archives of the Centre for Biotechnology. Graphic Design: Anne Asplund, Finepress Oy Printed by: Finepress Oy, Turku ISSN 1237-5217 CONTENTS Board of Trustees...................................................................... 2 Chairman’s Foreword................................................................. 3 From the Director....................................................................... 4 Year 2009 in a Nutshell.............................................................. 6 PhD and MSc Theses................................................................ 9 Funding..................................................................................... 10 Personnel 2009......................................................................... 11 The Finnish DNA Microarray Centre........................................... 14 The Proteomics Facility.............................................................. 16 Cell Imaging Core (CIC) ............................................................ 19 Virus Vector Facility . ................................................................. 22 Protein Crystallography Core Facility.......................................... 24 Data Mining and Modeling Group.............................................. 25 Protein Kinase Regulation of Brain Development and Disease......................................................... 29 Organisation of Neuronal Signaling Pathways............................ 33 Cytoskeletal and Survival Signaling............................................ 37 Cell Adhesion and Cancer......................................................... 42 Hypoxia in Cell Survival.............................................................. 45 Bioenergy Group....................................................................... 48 Kinetochore and Cancer Research Group.................................. 50 Canceromics Research Programme.......................................... 54 Signaling Pathways regulated by Oncogenic Pim Kinases......... 58 Molecular and Systems Immunology and Stem Cell Biology...... 61 Protein Crystallography.............................................................. 67 Bioinformatics Unit.................................................................... 72 Cell Fate ................................................................................ 76 Targeting Strategies for Gene Therapy....................................... 80 Transcriptional Regulation of Heat Shock Gene Expression....... 82 Cancer Cell Signaling................................................................. 86 Publications 2009...................................................................... 89 Life outside the Lab................................................................... 94 1 ORGANIZATION Board of Trustees 2009 Chairman HEINO Jyrki, Professor, University of Turku, Department of Biochemistry and Food Chemistry, Scientific Director, BioCity Turku Vice-chairman ERIKSSON John, Professor, Åbo Akademi University, Department of Biology Secretary LAHESMAA Riitta, Professor, Director, Turku Centre for Biotechnology Assistant Secretary JAAKKOLA Minttu, Coordinator, Turku Centre for Biotechnology and BioCity Turku Members ARO Eva-Mari, Academy Professor, the Academy of Finland, University of Turku, Department of Biology JALKANEN Sirpa, Professor, University of Turku, MediCity Research Laboratory JOHNSON Mark, Professor, Åbo Akademi University, Department of Biochemistry and Pharmacy KOUVONEN Ilkka, CEO, Turku Science Park Ltd LAHTI Reijo, Professor, University of Turku, Department of Biochemistry and Food Chemistry LASSILA Olli, Professor, University of Turku, Department of Medical Microbiology PYRHÖNEN Seppo, Professor, University of Turku, Department of Oncology SAXÉN Henrik, Vice Rector, Research Professor, Åbo Akademi University, Heat Engineering Laboratory TÖRNQUIST Kid, Professor, Åbo Akademi University, Department of Biology Vice-members ARO Hannu, Professor, University of Turku, Department of Surgery HINKKANEN Ari, Professor of Cell and Molecular Biology, Åbo Akademi University, Department of Biochemistry HUPA Leena, Lecturer, Åbo Akademi University, Åbo Akademi Process Chemistry PIISPANEN Tero, Project Manager, Turku Science Park Ltd SALAKOSKI Tapio, Professor, University of Turku, Department of Information Technology SISTONEN Lea, Academy Professor, the Academy of Finland SOUKKA Tero, Academy Research Fellow, Academy of Finland, University of Turku, Department of Biotechnology TOPPARI Jorma, Professor, University of Turku, Department of Physiology CHAIRMAN’S FOREWORD During the past years the expenses of the top-of-line instruments and the new research technologies have increased much faster than the budgets of the Universities. It is a well-known fact that the financing of the research infrastructure has become a major problem, not least in Finland. The six Finnish biocenters established in August 2006 a common organization, named as Biocenter Finland, to coordinate the development of national infrastructure and core facilities in life science area. Recently, the Finnish government decided to finance Biocenter Finland activities during the period 2010-2015 by 45 M€. The board of Biocenter Finland has already allocated most of its budget for different biocenters, including BioCity Turku. This funding allows Turku Centre for Biotechnology to renew its instrumentation in genomics, proteomics and bioimaging. Some funding has also been granted to bioinformatics, structural biology and virus vectors. Biocenter Finland financing will also be used to support core facilities in the Turku Center for Disease Modeling and the Turku PET centre. It is fair to say that without the support from Biocenter Finland it would have not anymore been possible to provide modern equipment and technologies for Finnish life scientists. Despite this positive development, the future of the research infrastructure remains to be a major concern in all Finnish biocenters, since the present support ends after year 2012. We should all work actively to secure the continuation of national infrastructure funding also after that. Turku Centre for Biotechnology has a long tradition in the building of high quality core facilities that have had numerous users also outside Turku. During the progression of the Biocenter Finland process the Turku core facilities will get even more important national role. It has been satisfying to realize that the organization and the administration of the core facilities are more advanced in Turku than anywhere else in Finland and that the working methods developed in Turku are often announced as ideal models for other biocenters. The beginning of the year 2010 brought remarkable changes in the Finnish university administration and implementation of new full-cost model in the research grants. For many scientists this has meant a huge increase in the number of hours used in the administration of the departments and the research projects. Similarly, the workload of the persons, who are directly involved in the maintenance of the departments has also dramatically increased. The complexity of the financial systems and the lack of proper computer programs have caused a lot of extra work and uncertainty. We can only hope that during the years to come the scientists will again be allowed to concentrate on research instead of administrative problems. Jyrki Heino, M.D., Ph.D., Professor of Biochemistry, Scientific Director of the BioCity Turku and Chairman of the Board of the Turku Centre for Biotechnology 2 3 FROM THE DIRECTOR Turku Centre of Biotechnology (CBT) was established in 1989 as a joint independent department of University of Turku and Åbo Akademi University. Today, it has evolved into a multidisciplinary research and service unit successfully crossing the boundaries of departments, faculties and even two universities! During the past 20 years Turku University and Åbo Akademi University have been centralizing demanding and expensive core facilities and research services to CBT to enable the most cost efficient use of the facilities for the local research community. CBT has developed into an internationally recognized research environment and today provides state-of the-art core facilities for altogether 80 BioCity Turku research groups and six research programs featuring altogether seven Academy of Finland Centers of Excellence. In June 2009 CBT celebrated its 20th anniversary with a symposium “From Cell Signaling to Medical Systems Biology” with participation of world class scientists many of whom are closely linked to CBT. The symposium dinner in Turku Castle was the highlight of the festivities. CBT has three key focus areas: 1) research, 2) education and training, and 3) providing core facilities and state-of–the-art technologies and research services in selected areas. The central areas of research are cell signaling, regulation of gene and protein expression and their interactions and systems biology. In 2009 we published 54 papers and 7 Ph.D.s were completed. Our group leaders were very active in organizing seminars, courses and symposia. Two new international group leaders were recruited. The Centre’s core facilities developed technology platforms and services in a close interaction with researchers. A significant investment has been made in developing the state-of-the-art platforms in genomics and functional genomics, proteomics, cell imaging and bioinformatics supporting the – omics technologies. The animal core facility takes up a sizeable portion of basic resources in serving researchers and Turku Centre for disease modelling. A joint organization of the Finnish biocenters, Biocenter Finland, was established in 2006 to facilitate national collaboration and to coordinate development of research infrastructures in Finland. In 2009, several of CBT group leaders worked intensively in various Biocenter Finland infrastructure networks and prepared for applications to develop infrastructures with funds Ministry of Education allocated to Biocenter Finland for this purpose. An international evaluation process resulted in substantial support to develop infrastructures, many of them at CBT. Hence, in addition to serving local needs, CBT now further develops and provides national services in several areas within the Biocenter Finland infrastructure network. Bioimaging and systems biology are the central infrastructures in the research strategy of Biocity Turku, and in the research strategy of both our universities. Vigorous development of technology platforms, research core facilities, and research infrastructures will continue, and support for both instrumentation and personnel will be provided. This extended collaborative network and available funding should provide us with new efficient ways to further develop the research infrastructure in Finland. 4 The universities prepared for going through a major change in the beginning of 2010. This included definition of new strategies for universities and Biocity Turku. Both the University of Turku and Åbo Akademi University chose to profile themselves as a research university, which is an excellent environment for CBT to build and develop on its strengths in research and research infrastructure. Moreover, molecular biosciences, CBT’s core competence is one of the central strategic strong areas of research. An important strategic goal of our universities is internationalization and international competitiveness. The universities strive towards strategic recruitment practice, to attract researchers with international merit. This has always been a practice at CBT. In fact, Finnish group leaders are a minority at our Centre. Our host universities’ commitment in internationalization will hopefully improve also availability of important administrative documents and meetings in English so that our foreign researchers and personnel can fully contribute to the development of our organizations. The past year was very successful. Active participation to Biocenter Finland infrastructure networks and numerous administrative challenges and changes increased the workload of our researchers as well as technical and administrative personnel. In spite of that, scientific achievements include several special awards and grants as well as papers published in top ranked journals by our scientists. It is my great pleasure to congratulate our scientists for their excellent achievements and thank everyone involved - including the CBT technical and administrative staff for their outstanding contributions. My special thanks to Dr. Eleanor Coffey, our current Vice Director, for taking the responsibility of directing and taking care of several duties of our Centre in 2009 and enabling me to carry out a sabbatical at Harvard Medical School. She took an extraordinary and outstanding care of the Centre. Thank you! Riitta Riitta Lahesmaa, M.D., Ph.D., Professor Director Turku Centre for Biotechnology University of Turku and Åbo Akademi University 5 YEAR 2009 IN A NUTSHELL RESEARCH AND EDUCATION 2009 • • • • • • • 54 scientific papers were published Seven new Ph.D.´s graduated CBT was awarded substantial funding (approximately 4 million €) from the Biocenter Finland infrastructure networks Two new international group leaders were recruited For undergraduate training, CBT again organized lecture courses and practical demonstrations including a laboratory course on “Functional Genomics” for Health Bioscience and Biology students (4 study points) and on “Medical Biotechnology” for Medical students (5 study points) The 20th anniversary was celebrated with a symposium “From Cell Signaling to Medical Systems Biology” 8 M.Sc. theses were completed DEVELOPMENT OF INFRASTRUCTURE, RESEARCH SERVICES AND CORE FACILITIES 2009 Finnish DNA Microarray Centre 2009 • • • • • The next-generation sequencing instrument: ABI SOLiD 3 was acquired and almost immediately upgraded to version 3Plus FDMC’s website was renovated to improve the visibility of the centre’s services Several events were organized in collaboration with various instrument manufacturers throughout the year to spread information on available and emerging technologies related to our services FDMC personnel gave talks in many scientific events and organized training courses to educate researchers on various topics Major efforts were carried out to further develop our project consultation services to help customers design their experiments. Also the centre’s bioinformatics team continued developing bioinformatics data analysis services Proteomics and Mass spectrometry Laboratory 2009 • • • • • 6 We became national coordinator of the new Proteomics and Metabolomics network of Biocentre Finland and secured BCF funding for personnel and instrumentation We coordinated three Nordic Networks on proteomics and mass spectrometry Several courses were organised nationally and internationally in collaboration with Nordforsk networks, FinnProt and the local universities QStar Elite: A second-hand Elite mass spectrometer was purchased and configured to run in-line with Dionex 3000 LC system Converted HCT Esquire (Ion trap, Bruker) for nanospray LC-MS/MS Cell Imaging Core 2009 • • • • • CIC was awarded substantial funding for in vivo and high resolution imaging from the Biocenter Finland Biological Imaging network CIC coordinated the Nordic Network on Imaging in Biology and Medicine with participation from Finland, Sweden, Norway, Russia, Denmark and Ireland. Network symposia and training events were held in Finland, Sweden, Ireland and Norway The Leica TCS-SP5 Stimulated Emission Depletion (STED) microscope for high resolution imaging were installed Daniel Abankwa was recruited from the Institute for Molecular Bioscience in Brisbane. He joined CBT as group leader and head of CIC CIC coordinated a plan to provide centralized, large scale data storage to the imaging community in Turku. This will be realized during 2010 in cooperation with Prof. Mark Johnson at Åbo Akademi Viral vector facility 2009 • • • • • Lenti vector production was added to its service repertoire Funding was secured from the Biocenter Finland Viral Gene Transfer infrastructure network to provide gene transfer tools to researchers Ketlin Adel was recruited as a new lab technician dedicated to service provision The Bio-safety level 2 lab was furnished with an inverted fluorescence microscope Jari Heikkilä, consultant on Lenti virus production, produced an exciting study on the use of oncolytic alphavirus for glioma treatment. This work was published in PLoS One in January 2010 Bioinformatics Unit 2009 • • • • • • The High-throughput bioinformatics group organized a two-day Chipster microarray data analysis course in collaboration with CSC High-throughput screening of natural molecules was established in conjunction with Prof. Pia Vuorela at Åbo Akademi University Increased collaboration between our unit and CSC in high power computing and cloud computing Participated in projects involving analysis of protein-protein and protein-ligand interactions, computer-aided prediction and intelligent molecular modeling and design; computerbased ligand docking and analysis; effects of molecular recognition and mutations on protein function Training of Ph.D. students in Bioinformatics and Computational Biology within the National Graduate School of Informational and Structural Biology. Contribution to courses organized by CBT and Åbo Akademi University Plans made to increase high-capacity storage infrastructure for archiving services to support Structural Bioinformatics, Structural Biology, Translational Area, Drug Discovery, Chemical Informatics and Bioimaging 7 Protein crystallography facility 2009 • • • • • Continued participation in several courses (Medical Biochemistry, TERBIO) and initiation of new ones (Protein Crystallography and Structural Genomics’,’How to solve a protein structure) with lectures and demonstrations in the X-ray facility Organised workshop ‘How to get the most from the protein structures’ New collaborative projects initiated with other groups in Finland and abroad All major crystallographic programs were kept upgraded to latest versions Funding received from Biocenter Finland towards a new X-ray generator Quality Assurance Unit 2009 • • • Organized courses for the university on (1) quality assurance and metrology and (2) how to assure the reliability of your laboratory test results Individual training for graduate and post-graduate students Carried out GLP inspections for the Central Animal Laboratory PhD and MSc Theses 2009 PhD Theses Name Supervisor Anckar, Julius Sistonen, Lea, Kaunisto, Aura Eriksson, John Kochin, Vitaly Eriksson, John Mattila, Elina Ivaska, Johanna Mialon, Antoine Westermarck, Jukka Pellinen, Teijo Ivaska, Johanna Åkerfelt, Malin Sistonen, Lea Site besides CBT ÅA/ Dept. of Biology UTU/ Dept. of Biology ÅA/ Dept. of Biology VTT/ UTU/Department of Medical Biochemistry and Genetics UTU/Department of Medical Biochemistry and Genetics VTT/ UTU/Department of Medical Biochemistry and Genetics ÅA/ Dept. of Biology MSc Theses Name Supervisor Männistö, Katja Koskinen, Päivi Lindquist, Julia Eriksson, John Rautoma, Karoliina Sistonen, Lea Rosenberg, Susanna Eriksson, John Sachin, Wakadkar Papageorgiou, Tassos Stykki, Heidi Eriksson, John Sun, Lihua Coffey, Eleanor Site besides CBT UTU/Dept. of Biology UTU/Dept. of Biology ÅA/ Dept. of Biology UTU/Dept. of Biology University of Skövde, Sweden UTU/ Dept. of Biology Tampere University of Technology CBT staff from left to right, front row: Virpi Korpiranta, Sarita Heinonen, Taina Kalevo-Mattila, Hannele Vuori, second row: Marjo Hakkarainen, Riitta Lahesmaa, Riina Plosila, Aila Jasmavaara, Susanna Pyökäri, Eva Hirvensalo, Sirkku Grönroos, back row: Juha Strandén, Jouko Sandholm, Markku Saari, Pasi Viljakainen, Mikael Wasberg, Satu Alanko, Perttu Terho, Petri Vahakoski, Mårten Hedman, Petri Kouvonen, Arttu Heinonen. 8 9 FUNDING PERSONNEL 2009 Sources of funding received by Centre for Biotechnology in 2009 (9.4 Million €) Academy of Finland 15% Tekes 3% Others 12% EU 13% Services 20% Universities 37% Administration LAHESMAA Riitta, Director, Professor, Group Leader GRÖNROOS Sirkku, Senior Administrative Assistant HEINO Ilona, Student HIRVENSALO Eva, Clerical Official JAAKKOLA Minttu, Coordinator JASMAVAARA Aila, Clerical Official PLOSILA Riina, Coordinator BioCity Turku HEINO Jyrki, Biocity Turku Scientific Director, Professor JAAKKOLA Minttu, Coordinator Technical Staff ANDERSEN Raija, Laboratory Technician HEDMAN Mårten, Systems Manager KORPIRANTA Virpi, Instrument Maintenance SARA Rolf, WASBERG Mikael, Laboratory Manager SOITAMO Ann-Christine, Student STRANDÉN Juha, Laboratory Engineer VAHAKOSKI Petri, Systems Manager VILJAKAINEN Pasi, Senior Technician VUORI Hannele, Instrument Maintenance Data Mining and Modeling AITTOKALLIO Tero, Group Leader, Adjunct Professor ELO Laura, Postdoctoral Fellow GAO Bin, Undergraduate Student HIISSA Jukka, Graduate Student JÄRVINEN Aki, Undergraduate Student KOSKINEN Ville, Undergraduate Student LAAJALA Essi, Undergraduate Student LAAJALA Teemu Daniel, Undergraduate Student LINDEN Rolf, Graduate Student NATRI Lari, Undergraduate Student OKSER Sebastian, Graduate Student PIIPPO Mirva, Undergraduate Student SALMELA Pekka, Undergraduate Student SALMI Jussi, Postdoctoral Fellow TUIKKALA Johannes, Graduate Student VÄHÄMAA Heidi, Graduate Student Protein Kinase Function in Brain Development and Disease COFFEY, Eleanor, Group Leader, Academy of Finland Research Fellow BIALEK Agnieska, Undergraduate Student DESHPANDE Prasannakumar, Graduate Student HEALY Mary-Ann, Undergraduate HEIKILÄ. Hanna, Undergraduate Student KOMULAINEN, Emilia, Graduate Student MOHAMMAD Hasan, Graduate Student MYSORE, Raghavendra, Graduate Student PADZIK, Artur, Graduate Student Pyökäri Susanna, Laboratory Technician SUN Lihua, Graduate Student TUITTILA Minna, Postdoctoral Researcher 10 WANG Yubao, Graduate student ZDROJEWSKA, Justyna, Graduate student Proteomics and Mass Spectrometry CORTHALS Garry, Group Leader, Head of Proteomics ANDERSÉN Raija, Laboratory Technician CARREIRA DOS SANTOS Hugo Miguel Baptista, Visiting Graduate Student HEINONEN Arttu, Project Engineer KANNASTE Olli, Undergraduate Student KAUNISMAA Katri, Undergraduate Student KOUVONEN Petri, Researcher RALPH Eliza, Systems Administrator ROKKA Anne, Postdoctoral Fellow SUNI Veronika, Graduate Student VEHMAS Anni, Undergraduate Student NEES Susanne, Coordinator SAEDI Firouz, Undergraduate student Organisation of Neuronal Signaling Pathways COURTNEY Michael, Professor, Group Leader HO, Franz, Postdoctoral Researcher LI, Lili, Graduate Student LIU, Xiaonan, Graduate Student LOPEZ RODRIGUES, Maykel, Graduate Student MARTINSSON, Peter, Postdoctoral Researcher PEVGONEN, Veera, Technicain TUITTILA Minna, Postdoctoral Researcher VERGUN, Olga, Postdoctroal Researcher WANG, Xijun, Graduate Student YADAV, Leena, Graduate Student Protein Phosphorylation Group ERIKSSON John, Group Leader, Professor ASAOKA Tomoko, Graduate Student FERRARIS Saima, Graduate Student HYDER Claire, Graduate Student IMANISHI Susumu, Postdoctoral Fellow KASTU Juha, Project Engineer KAUNISTO Aura, Post-doctoral Fellow KOCHIN Vitaly, Graduate Student LAZARO, Glorianne, Exchange Student LINDQVIST Julia, Graduate Student ISONIEMI Kimmo, Graduate Student PALLARI Hanna-Mari, Graduate Student PEUHU Emilia, Graduate Student REMES Mika, Graduate Student SAARENTO Helena, Research Associate SÖDERSTRÖM Thomas, Post-doctoral fellow TORVALDSON Elin, Graduate student Cell Imaging Core COFFEY Eleanor, Academy of Finland Research Fellow, Coordinator of the Cell Imaging Unit ERIKSSON John, Group Leader, Professor 11 KORHONEN Jari, Project Engineer SANDHOLM Jouko, Research Engineer TERHO Perttu, Project Engineer Cell Adhesion and Cancer IVASKA Johanna, Group Leader, Professor ARJONEN Antti, Undergraduate Student HÖGNÄS Gunilla, Undergraduate Student MARTTILA Heidi, Laboratory Technician MAI Anja, Graduate student MATTILA Elina, Graduate student NEVO Jonna, Graduate Student PELLINEN Teijo, Graduate student SIIVONEN Jenni, Laboratory Technician TUOMI Saara, Graduate Student VELTEL Stefan, Postdoctoral Fellow VUORILUOTO Karoliina, Graduate Student Hypoxia Group JAAKKOLA Panu, Group Leader, Adjunct Professor, Academy Research Fellow, the Academy of Finland HEIKKINEN Pekka, Graduate Student HIMANEN Virpi, Undergraduate Student HÖGEL Heidi, Graduate Student JOKILEHTO Terhi, Graduate Student KALEVO-MATTILA Taina, Laboratory Technician KULJU Tuomas, Undergraduate Student NUMMELA Marika, Graduate Student PURSIHEIMO Juha-Pekka, Postdoctoral Fellow RANTANEN Krista, Graduate Student RÄMÖ Olli, Undergraduate Student VUORINEN Raisa, Laboratory Technician Kinetochore and Cancer Research Group KALLIO Marko, Group Leader, Senior Research Scientist, Adjunct Professor AHONEN Leena, Postdoctoral Fellow HALONEN Tuuli, Undergraduate Student JAAKKOLA Kimmo, Postdoctoral Fellow KUKKONEN-MACCHI Anu, Graduate Student MÄKI-JOUPPILA Jenni, Graduate Student NARVI Elli, Postdoctoral Fellow OETKEN-LINDHOLM Christina, Postdoctoral Fellow SALMELA Anna-Leena, Graduate Student TOIVONEN Pauliina, Laboratory Technician WINSEL Sebastian, Postdoctoral Fellow VUORILUOTO Mariaana, Graduate Student 12 Canceromics Reasearch Programme KALLIONIEMI Olli, Group Leader, Director PLOSILA Riina, Coordinator AAKULA Anna, Graduate Student BUCHER Elmar, Graduate Student BJÖRKMAN Mari Graduate Student GUPTA Santosh, Graduate Student KETOLA Kirsi, Graduate Student KOHONEN Pekka, Graduate Student POLLARI Sirkku, Graduate Student VAINIO Paula, Graduate Student Signaling Pathways Regulated by Oncogenic Pim Kinases KOSKINEN Päivi, Group Leader, Adjunct Professor EEROLA Sini, Undergraduate Student LAITERÄ Tiina, Undergraduate Student MÄNNISTÖ Katja, Undergraduate Student RAINIO Eeva-Marja, Postdoctoral Fellow SANDHOLM Jouko, Graduate Student SANTIO Niina, Undergraduate Student VAHAKOSKI Riitta, Graduate Student VIRTANEN Juho, Undergraduate Student Molecular Immunology Group LAHESMAA Riitta, Director, Professor, Group Leader AHLFORS Helena, Graduate Student FILEN Sanna, Graduate Student GUPTA Bhawna, Postdoctoral Fellow HAKKARAINEN Marjo, Laboratory Technician HEINONEN Mirkka, Graduate Student HEINONEN Sarita, Laboratory Technician JÄRVENPÄÄ Henna, Graduate Student KORHONEN Juha, Graduate Student KUMAR Sunil, Postdoctoral Fellow KYLÄNIEMI Minna, Graduate Student LAHTI Essi, Undergraduate Student LUND Riikka, Postdoctoral Fellow LÖNNBERG Tapio, Graduate Student MOULDER Robert, Senior Scientist NYSTRÖM Joel, Undergraduate Student NÄRVÄ, Elisa, Undergraduate Student RAHKONEN Nelly, Undergraduate Student RASOOL Omid, Adjunct Professor, Senior Scientist SALONEN Verna, Graduate Student TAHVANAINEN Johanna, Graduate Student TRIPATHI Subhash, Graduate Student TUOMELA Soile, Graduate Student Quality Assurance Unit LINKO Linnéa, Group Leader, Adjunct Professor Computational Systems Biology ORESIC Matej, Group Leader, Adjunct Professor, Chief Research Scientist KATAJAMAA Mikko, Graduate Student LÖNNBERG Tapio, Graduate Student Protein Crystallography PAPAGEORGIOU Tassos, Group Leader, Adjunct Professor AGIUS Jeremie, Exchange Student ANDERSSON Charlotta, Graduate Student CHOUHAN Bhanupratap Singh, Undergraduate student DHAVALA Prathusha, Undergraduate Student DUBREUIL Christine, Student HAIKARAINEN Teemu, Graduate Student HAVUKAINEN Heli, Graduate Student KAUKO Anni, Postdoctoral Fellow ROUE Carole, Undergraduate Student SAARINEN Susanna, Graduate Student WECKSTRÖM Kristian, Senior Scientist Bioinformatics Unit DENESSIOUK Konstantin, Group Leader (Structural Bioinformatics) GYENESEI Attila, Group Leader (High-throughput Bioinformatics) CHOUHAN Bhanupratap Singh, Graduate Student JUNTTILA Sini, Graduate Student LAIHO Asta, Project Engineer KYTÖMÄKI Leena, Undergraduate Student TAMMINEN Seppo, Undergraduate Student Cell fate SAHLGREN Cecilia, Group Leader, Academy of Finland Research Fellow MAMAEVA Veronika, Postdoctoral Fellow HIETAMÄKI Marika, Graduate Student LANDOR Sebastian, Graduate Student BATE-EYA Laurel Tabe, Graduate Student ANTFOLK Daniel, Undergraduate Student ANTILA Christian, Undergraduate Student GRANQVIST Cecilia, Undergraduate Student NIEMI Rasmus, Undergraduate Student SAARENTO, Helena, Laboratory Technician Targeting Strategies for Gene Therapy SAVONTAUS Mikko, Group Leader, Adjunct Professor EEROLA Kim, Graduate Student MATTILA Minttu, Undergraduate Student TOIVONEN Raine, Graduate Student Transcriptional Regulation of Heat Shock SISTONEN Lea, Group Leader, Professor, Academy Professor until July 2009 AALTO Anna, Undergraduate Student AHLSKOG Johanna, Graduate Student ANCKAR Julius, Postdoctoral Fellow BERGMAN Heidi, Undergraduate Student BJÖRK Johanna, Graduate Student BLOM Malin, Undergraduate Student BLOMSTER Henri, Graduate Student BUDZYNSKI Marek, Graduate Student CHITIKOVA Zhanna, Graduate Student ELSING Alexandra, Graduate Student HENRIKSSON Eva, Postdoctoral Fellow KARLBERG Henrica, Undergraduate Student RAUTOMA Karoliina, Undergraduate Student SAARENTO Helena, Research Associate SANDQVIST Anton, Graduate Student SIIMES Jenny, Undergraduate Student VARTIAINEN Aki, Undergraduate Student VIHERVAARA Anniina, Graduate Student ÅKERFELT Malin, Postdoctoral Fellow Finnish DNA Microarray Centre GYENESEI Attila, Group Leader SARA Rolf, Technical Team Leader, Laboratory Manager Juha-Pekka Pursiheimo, Senior Scientist HEINONEN Tiia, Laboratory Technician JUNNI Päivi, Laboratory Technician NURMI Miina, Laboratory Technician RISSANEN Oso, Laboratory Technician SIPILÄ Anna, Undergraduate Student VENHO Reija, Laboratory Technician VIRTANEN Eveliina, Project Engineer KYTÖMÄKI Leena, Biotechnology Engineer LAIHO Asta, Project Engineer TAMMINEN Seppo, Undergraduate Student JUNTTILA Sini, Project Engineer Cancer Cell Signaling WESTERMARCK Jukka, Group Leader, Professor, CÕME Christophe, Postdoctoral Fellow HALONEN Tuuli, Graduate Student KALEVO-MATTILA Taina, Laboratory Technician KAUR Amanpreeet, Graduate student LAINE Anni, Graduate Student MANNERMAA Leni, Scientist MIALON Antoine, Graduate Student NIEMELÄ Minna, Graduate Student OKKERI Juha, Postdoctoral fellow POKHAREL Yuba, Postdoctoral fellow VENTELÄ Sami, Postdoctoral Fellow Biobanking and Biomolecular Resources Research Infrastructure (BBMRI) VUORIO Eero, BBMRI Executive Manager, Chancellor SALMINEN-MANKONEN Heli, Adjunct professor, Project manager GRÖNROOS Sirkku, Project assistant 13 The Finnish DNA Microarray Centre http://fmsc.btk.fi Head: Attila Gyenesei, Ph.D., Senior Scientist, Group leader Contact information: Turku Centre for Biotechnology, BioCity, Tykistökatu 6A, P.O. Box 123, FIN-2050 Turku, Finland. Tel. +358-2-333 8634 Fax +358-2-333 8000. Email: [email protected] Personnel: Tiia Heinonen, Päivi Junni, Leena Kytömäki, Asta Laiho, Miina Nurmi, Oso Rissanen, Rolf Sara, Reija Venho, Eveliina Virtanen, JuhaPekka Pursiheimo, Riikka Lund, Sini Junttila, Seppo Tamminen, Sarita Heinonen, Ritva Ala-Kulju, Anna Sipilä General description: The Finnish DNA Microarray Centre (FDMC) is an internationally recognised Functional Genomics Core Facility that is part of the Turku Centre for Biotechnology. As a national core facility, we provide state-of-the-art research technologies and services in the areas of genomics, epigenomics, transcriptomics and bioinformatics for the Finnish as well as the international scientific community. Our services include next-generation sequencing, microarray based services (gene expression analysis, exon-specific expression analysis, miRNA analysis, ChIP-on-chip analysis, aCGH and SNP genotyping), Real-Time PCR and traditional DNA sequencing. Our service covers all steps from experimental planning and design to sample processing and bioinformatics data analysis. The centre also regularly organizes courses, symposia and training for its users. FDMC hosts commercial microarray platforms for genome-wide RNA expression profiling, SNP genotyping and comparative genomic hybridization needs. These platforms include Affymetrix GeneChip©, Illumina Sentrix Bead Array© and Agilent DNA technology services for all of which we have been granted the Certified Service Provider status. All platforms have dedicated scanners and software for array data analysis. Diverse aspects of the microarray techniques are continuously developed and tested. During 2009 the Centre acquired a high-throughput nextgeneration sequencing (NGS) instrument SOLiDTM 3Plus from Applied Biosystems. The system supports a wide range of genetics applications covering genomics, transcriptomics and epigenomics and it is distinguished by its unmatched accuracy and capability of generating more than 60 gigabases of map-able sequence data per run. The combination of increased throughput, shorter run times, and improved data analysis make the SOLiD technology an ideal choice for research applications in any genetics project. The system enables for example distinguishing strand specific expression patterns, discovery of novel transcripts and splice variations without the bias of microarrays, detecting SNPs at low coverage with a low false positive rate, global assessment of DNA-protein binding interactions and characterization of structural rearrangements including balanced translocations. The Microarray Centre offers a number of other genomic analysis technologies for gene expression, SNP and genotyping studies including a sequencing facility and real-time PCR service. Services include BioRad Experion runs for verifying the RNA quality. Bioinformatics data analysis and data mining are included in the data analysis service that is provided for microarray and nextgeneration sequencing customers. The data handling is done by our bioinformaticians, using both commercial and R/Bioconductor software tools. Seminars and practical courses on microarrays and related bioinformatics are held frequently to facilitate knowledge transfer within the field, often this is done in collaboration with graduate schools. Funding: Ministry of Education and the Centre of Expertise of Southwest Finland Users: Finnish DNA Microarray Centre has customers from Finnish universities, biocenters and research institutes in the field of biosciences as well as local companies and foreign companies and universities - altogether from more than 200 research groups. Illumina Sentrix Bead Array© service was started with whole human and mouse genome transcriptomic profiling and from the beginning of 2007 rat arrays have also been offered for gene expression studies. Illumina’s GoldenGate Assay protocol provides high-quality and high-multiplex genotyping. Affymetrix GeneChip© service has provided whole transcript expression arrays for human, mouse and rat, and expression arrays for a wide selection of organisms. Platform also provides many arrays for SNP detection and copy number variation. Agilent DNA technology service has provided whole genome expression arrays, which are designed and validated for onecolor and two-color processing. From 2007 these have been also available as multi-packs. Beside catalog arrays also custom arrays have been processed. Also micro RNA arrays are available. 14 From left to right: Leena Kytömäki, Sini Junttila, Päivi Junni, Leni Mannermaa, Sanna Vuorikoski, Attila Gyenesei, Riikka Lund, Juha-Pekka Pursiheimo, Eveliina Virtanen, Reija Venho, Sarita Heinonen. 15 THE PROTEOMICS FACILITY http://proteomics.btk.fi/ Head: Garry Corthals, Ph.D. (2005). Address: Turku Centre for Biotechnology, BioCity, Tykistökatu 6, P.O. Box 123, FI-20521 Turku, Finland. Tel. +358-2-333 8889, Fax. +358-2-333 8000. Email: [email protected] Personnel: Senior scientists: Dr. Anne Rokka, Ph.D.; Associate scientist: Petri Kouvonen, M.Sc. (Ph.D. student); Laboratory Engineer: Arttu Heinonen; Technician: Raija Andersen; Bioinformatician: Eliza Ralph Steering Committee: Prof. Eva-Mari Aro (University of Turku), Dr. Eleanor Coffey (Åbo Akademi University), Prof. John Eriksson (Åbo Akademi University), Prof. Jyrki Heino (University of Turku), Prof. Riitta Lahesmaa (CBT), Prof. Matti Poutanen, Prof. Craig Primmer (University of Turku), Prof. Jukka Westermarck (CBT) and Prof. Johanna Ivaska (VTT & CBT) General description: The facility aims to support life science research in need of proteomics and MS services. Key questions revealed by proteomics technologies are differences in protein abundance, measurement of post translational modifications, defining protein complexes that carry out specific functions and localisation of proteins in tissues. To address these questions, the facility develops and innovates, in collaboration with national research groups, new methodologies in proteomics. Most services provided by the facility focus on mass spectrometric strategies integrated with protein and peptide enrichment workflows for large-scale quantitative analysis of proteomes or detailed characterisation of single proteins, such as their state of phosphorylation. In recent years the facility has developed a wide basis of operation and expertise, which are: Quantitative proteomics – several groups are independently using labelling iTRAQ and more recently SILAC based quantitation. In addition, several projects are providing a framework for development of label-free quantitative analysis, especially useful for clinical samples. Post-translational modifications – several groups have now published new tools in analysis of protein phosphorylation and sumoylation. Mass spectrometry imaging – MS imaging continues to develop locally, with expansion of the activities taken up in the new Master’s Degree Programme in Biomedical Imaging. Biological mass spectrometry – numerous groups use the Facility for additional analytical services that are not directly related to proteomics, such as protein, peptide and small molecule structure determination, mass determination and peptide and protein purity analysis. Protein separation – by liquid chromatography and a variety of gel based methods such as 1-DE, 2-DE, peptide-IPG and blue native gel electrophoresis. Bioinformatics – identification, quantitation and validation studies, reporting and software development. 16 From left to right: Anne Rokka, Arttu Heinonen, Petri Kouvonen, Eliza Ralph. 17 The core operations of the facility are in services and training in proteomics to the local and national biosciences community. In doing so the facility aims to cover diverse fields requiring the analysis of proteins in cells, tissues, organisms and body fluids. To achieve this, the facility is active in the following three activities: Analytical services The aim is to offer the best possible analytical proteomics services to bioscience researchers in academia and industry, both locally and nationally through Biocentre Finland coordinated activities. Locally, a broad range of MS analyses are performed, whilst at the national level the facility spearheads developments in two of its services; MS-based quantitative proteomics and phosphorylation determination. Training Our aim is to organise and coordinate of training events and symposia focusing on Proteomics, mass spectrometry and computational tools and procedures for the analysis of MS data. Innovation The facility develops tools and procedures in collaboration with facility users and groups. Through activities linked to Biocenter Finland services, two focus areas of development for the facility are in quantitation and phosphorylation analysis. The facility has excellent instrumentation for a wide range of MS analyses, including four different mass spectrometers: an Ultraflex II (MALDI-TOF/TOF), a QStar Pulsar and Elite (ESI-QqTOF) and a Esquire HTC (ESI-ion trap MS). The ESI instruments are all in-line coupled to nanoHPLC systems, and can also be used for direct nanospray ESI. Additional nanoHPLC systems robotic microfraction collectors enable real-time collection of peptide fractions, direct on MALDI targets. A LIMS system and computational analysis pipeline. The facility also hosts hotel services where visiting scientists can use shared facility space for sample preparation and analysis. In addition to analytical services, the facility organises and participates in national collaborative programs and research efforts with others throughout the Nordic region and worldwide. The facility also participates actively in national and international educational programs in Europe and abroad, such as Nordforsk, EuPA and HUPO. Funding: Biocenter Finland, The Academy of Finland, TEKES, City of Turku, Ministry of Education/Proteomics and the Centre of Expertise of Southwest Finland, Turku University and Åbo Akademi University, Bruker Daltonics, the Systems Biology Research Program. Users: Biocentre Finland universities, The University of Turku, Åbo Akademi University, Turku Polytechnic. CoE’s in: Translational Genome-Scale Biology; Evolutionary Genetics and Physiology; Integrative Photosynthesis and Bioactive Compound Research at Systems Biology Level; and Åbo Akademi CoE in Cell Stress. The Systems Biology Research Program, national research groups, Turku Hospitals, the Finnish Red Cross and the National Animal Research Centre. 18 CELL IMAGING CORE (CIC) http://[email protected]/ Coordinator and group leader: Eleanor Coffey, Ph.D., Adjunct Professor in Cellular and Molecular Biology, Turku Centre for Biotechnology, BioCity, 5th floor, Tykistökatu 6, FI-20521, Finland. Tel. +358-2-3338605, Fax. +358-2-3338000. Email: [email protected] Technical Team: Jouko Sandholm, M.Sc., Senior Researcher Microscopy, Email: [email protected], Perttu Terho, B.Sc., Technical Engineer Flow Cytometry, Email: [email protected] Imaging Consultants: Prof. Michael Courtney, Ph.D., Research Director, A.I.Virtanen Institute, University of Kuopio, Finland. Email: [email protected] Kaisa Heiskanen, Ph.D., Orion Pharma. Email: kaisa.heiskanen@ orionpharma.com Steering Committee: Prof. Olli Carpén, M.D., Ph.D., Prof. John Eriksson (chairman), Ph.D., Prof. Jyrki Heino, M.D., Ph.D., Prof. Pekka Hänninen, Ph.D., Prof. Sirpa Jalkanen, M.D., Ph.D., Prof. Riitta Lahesmaa, M.D., Ph.D., Prof. Olli Lassila, M.D., Ph.D., Prof. Matti Poutanen, Ph.D., Prof. Lea Sistonen, Ph.D., Kid Törnquist, Ph.D. The Cell Imaging Core (CIC) is a centralised facility that coordinates biological imaging activities between the University of Turku, Åbo Akademi University, VTT Technical Research Centre, Turku Functional Genomics Centre, the National Centre for Disease Models and the biophysics community, while providing services at the national level. The mission of CIC is to provide state-of-theart cell imaging and cell sorting technologies and to make them available to academic and industrial researchers. The primary goal of CIC is to enhance the research and teaching environment of BioCity Turku. To help meet these goals, the core unit • provides technical training to local and visiting researchers and to industries • offers consultation on experimental design and image analysis • evaluates new methods and fluorescence tools and communicates acquired knowledge to users • implements advances in hardware and software relevant for biomedical sciences • provides ongoing education in theory and practice by organising training courses and international workshops As a result, CIC has grown in significance, the services provided being instrumental to the publication of over 200 international scientific articles in recent years, many of which are in the most prestigious journals. Our staff include a coordinator and experienced applications specialists who maintain the instruments, learn new technologies and most importantly, provide personal training to users. Our areas of technical expertise are confocal microscopy 19 University Medical School, Chicago), Irina Majoul (Royal Holloway, London), Scott Brady (University of Illinois at Chicago), Gyorgy Hajnoczky (Thomas Jefferson University, Philadelphia), Michael Courtney (University of Kuopio), Teng Leong-Chew (Northwestern University Medical School, Chicago), Stephen Ogg (Institute of Medical Biology, Singapore), Kota Miura (EMBL, Heidelberg), Martin Leahy (University of Limerick), Gregory McNerny (Univeristy of California Davis). The CIC-initiated Nordic Network on Imaging in Medicine and Biology adds to this list a range of additional participants, principally from the Nordic area. Ongoing intimate collaboration between these experts and the Cell Imaging Core ensures continued transfer of advanced imaging skills from these leading labs. To strengthen bioscience imaging in the Turku region, CIC has expanded its web pages to include imaging technologies and contacts across the campus. CIC maintains close contact with imaging specialists locally and is a major player in the Turku Bioimaging initiative, an active discussion forum established in 2007 to promote imaging in the Turku region. From left to right: Jouko Sandholm, Eleanor Coffey, Perttu Terho, Markku Saari. (including timelapse and spectral detection), widefield fast CCD imaging, laser microdissection, high throughput cell sorting and advanced flow cytometry software development. To maintain the advanced, national level service provided so far, we organise training programmes, service existing equipment, sustain research on new imaging techniques, and implement the latest technological advances demanded by the Finnish research community. CIC has succeeded both as a service provider and as a point of integration of emerging imaging technologies. Added value is achieved by the present local expertise in the Turku area in the fields of fluorescence-activated cell sorting, confocal microscopy, fluorescence-based screening and robotic instrumentation, imaging-based high-content screening, in vivo animal imaging, and viral gene transfer, and by the presence of the National Centre for Disease Models and the transgenic animal core facility. CIC coordinates the Nordic Network on Imaging in Medicine and Biology. This network gathers the leading units on cellular and medical imaging in the Nordic region and was funded initially by the Joint Committee of the Nordic Research Councils and currently by Nordforsk, the funding agency of the Nordic Council of Ministers. The network assembles 38 prominent research groups and aims to improve interdisciplinary training and cooperation between diverse imaging fields. The following international leaders in the field have contributed to advanced imaging education in Turku providing during their visits practical as well as theoretical training: Jennifer LippincottSchwartz (NIH, USA), Stefan Hell (Max Planck Institute for Biophysical Chemistry, Gottingen University, Germany), Rainer Duden (Royal Holloway, London), Robert Goldman (Northwestern 20 21 VIRUS VECTOR FACILITY http://virusvec.btk.fi Coordinator: Eleanor Coffey, Ph.D., Adjunct Professor in Cellular and Molecular Biology, Turku Centre for Biotechnology, BioCity, 5th floor, Tykistokatu 6, FI-20521, Finland. Tel. +358-2-3338605, Fax. +3582-33378000. Email: [email protected] To build on local expertise in gene transfer technologies, the Virus Vector Facility networks with experts in viral vector design. Thus a number of local experts on retroviruses and alpha-viruses are available for consultation on vector design, production and concentration. Research, Development and Training: Anna Cvrljevic, postdoctoral researcher (Westermarck lab), Turku Centre for Biotechnology, BioCity 5th floor, Tykistökatu 6, FI-20521, Finland. Email: [email protected] Technical Team: Marjo Hakkarainen, Laboratory Technician, Email: [email protected] Susanna Pyökäri, Laboratory Technician, Email: susanna.pyökä[email protected] Ketlin Adel, Laboratory Technician, Email: [email protected] The Virus Vector Facility produces viral vectors for local and national research groups. During 2009, the Virus Vector Facility joined the national infrastructure network on Viral Gene Transfer, funded by Biocenter Finland. Our primary function is to facilitate the use of viral vectors by national researchers. To meet these goals the virus vector facility • Produces on demand adenoviruses and lentiviruses expressing genes of interest, as a research service • provides a fully equipped bio-safety level-2 lab for researchers wishing to produce their own vectors (replication deficient viruses only) • supplies working protocols for production of adeno and lenti vectors and trains researchers in the safe preparation and handling of viral vectors • organises seminars and courses emphasizing practical issues related to gene transfer technology • coordinates a network of local experts from whom consultation on design of viral vectors can be sought The virus vector facility has a national user base with regular customers from the universities of Turku, Oulu and Helsinki as well as customers from biotech companies. In addition to customer service, our infrastructure is used by 16 local research groups producing adenoviruses, adeno-associated virus, retro- and lentivirus for their own research purposes. These viruses are typically used to obtain high efficiency gene transfer in difficult to transfect cells such as primary cultures of T lymphocytes and neurons and for in vivo cancer studies. Another typical application is the use of viral vectors for delivery of shRNA. Protocol optimization has been completed for high efficiency gene silencing in primary cultured neurons and T lymphocytes and more recently investigators are using virally delivered miRNAs for gene knockdown studies. 22 From left to right: Susanna Pyökäri, Anna Cvrljevic, Jukka Westermarck, Marjo Hakkarainen, Eleanor Coffey. 23 PROTEIN CRYSTALLOGRAPHY CORE FACILITY DATA MINING AND MODELING GROUP http://crystal.btk.fi http://users.utu.fi/teanai/ Head: Anastassios C. Papageorgiou, Ph.D., Adjunct Professor in Biochemistry and Structural Biology Turku Centre for Biotechnology, BioCity, Tykistökatu 6A, FI-20521 Turku, Finland. Tel. +358-2-3338012, Fax +358-2-3338000. Email: [email protected] Principal investigators: Tero Aittokallio, Ph.D., Docent in Biomathematics, Department of Mathematics, University of Turku, FI-20014 Turku, Finland. Tel. +358-2-3336030, Fax. +358-2-3336595. Email: [email protected] Technical Team: Technical support: Juha Strandén, Pasi Viljakainen Computational support: Petri Vahakoski, Mårten Hedman Steering committee: Jyrki Heino, Professor, Department of Biochemistry and Food Chemistry, University of Turku Reijo Lahti, Professor, Department of Biochemistry and Food Chemistry, University of Turku Tiina Salminen, Senior lecturer, Department of Biochemistry, Åbo Akademi Description of the Facility X-ray crystallography is a proven technique for detailed structurefunction studies of biological macromolecules. The Protein Crystallography Core Facility at CBT uses state-of-the-art equipment to determine the crystal structures of various proteins and their complexes. The Facility consists of an X-ray generator, Mar345 imaging plate detector, Osmic confocal mirrors, a Cryostream Cooler (Oxford Cryosystems) and several computers running under Unix or Linux operating systems for heavy duty calculations. The Unit has several workstations to run variety of graphic software (O, XtalView, Grasp, COOT, CCP4mg, PyMol), modeling and docking programs (MODELLER, Hex, Discovery Studio), and various crystallographic packages (HKL, CNS, CCP4, SHELX, SOLVE, SHARP, PHENIX) for data processing, analysis, phasing and refinement. The Facility has long expertise in all steps of a crystal structure determination: protein purification, crystallization, data collection (both in-house and in synchrotron radiation sources), data processing, phase determination, refinement and detailed analysis of the final structure. Incubators at different temperatures (4°C, 16°C and 23°C) for crystallization set-ups and a number of commercial screens for establishing initial crystallization conditions are available. In addition, we can provide homology modeling services and design of mutants for functional studies as well as ab initio predictions of protein structures. Since protein crystallography requires highly pure protein prepapations, we can offer full support and consultation on protein purification strategies apart from the services in structure determination and modeling. The Unit is able to undertake research projects for academic groups and companies, either in the form of collaborative efforts or as services. Protein Crystallography requires a multi-disciplinary approach and we are especially interested in bringing together expertise from various groups in order to better understand the structure-function relationship of biological macromolecules in key biological processes. Funding: Systems Biology research program, Biocenter Finland 24 Olli Nevalainen, Ph.D., Professor of Computer Science, Turku Centre for Computer Science, Joukahaisenkatu 3-5 B, FI-20520 Turku, Finland. Tel. +358-2-3338631; Email: [email protected] Biographies: Tero Aittokallio received his Ph.D. in Applied Mathematics from the University of Turku in 2001. In 2006-2007, he was a postdoctoral research fellow in the Systems Biology Group at Institut Pasteur, Paris. Currently he is an Academy Research Fellow in the Biomathematics Research Group. Olli S. Nevalainen received his Ph.D. degree in 1976. From 1972 to 1976, he was a lecturer with the Department of Computer Science, University of Turku. From 1976 to 1999, he was an Associate Professor, and since 1999 a Professor in the same department. Personnel: Post-doctoral researchers: Laura Elo, Ph.D., Jussi Salmi, Ph.D., Graduate students: Bin Gao, M.Sc., Jukka Hiissa, M.Sc., Ville Koskinen, M.Sc., Rolf Linden, M.Sc., Sebastian Okser, M.Sc., Johannes Tuikkala, M.Sc., Heidi Vähämaa, M.Sc., Undergraduate students: Aki Järvinen, Essi Laajala, Teemu Daniel Laajala, Lari Natri, Mirva Piippo. Description of the project: The research group develops mathematical modeling methods and implements computational analysis tools for mining data generated by modern high-throughput biotechnologies. The large number of components probed together with high technical and biological variability can make it difficult to extract pertinent biological information from the background noise. This has increased the need for computational models and tools that can efficiently integrate, visualize and analyze the experimental data so that the most important questions can be addressed and the meaningful interpretations can be made. The eventual aim is to model and explain the observations as a dynamic interaction of key molecular components and mechanisms controlling the underlying system. Data mining protocols developed so far cover a wide range of highthroughput biotechnologies, such as gene and exon arrays (cDNA, Affymetrix and Illumina platforms) for global gene expression profiling, together with RNA interference (RNAi) and chromatin immunoprecipitation (ChIP) studies (ChIP-chip and ChIP-seq) for monitoring transcriptional regulation on a global scale, as well as mass-spectrometry (MS)-based assays for large-scale proteomic studies and comparative genomic hybridizations (CGH) for detecting gene amplification or deletion events. One of the most important computational challenges is to take full advantage of all the accumulated data, both from own laboratory and from public 25 repositories, to obtain a more comprehensive view of the system under study. We are developing a data integration approach, which can effectively correct for the technical variation characteristic to various experimental platforms, and hence improve the comparability of different experiments, identification of differentially expressed genes and proteins, and inference of their interaction partners in global cellular networks. Such integrative network-based modeling approach can provide robust and unbiased means to reveal the key molecular mechanisms behind the systems behavior and to predict its response to various perturbations. In clinically-oriented research, the modeling approach has the potential to improve our understanding of the disease pathogenesis and help us to identify novel molecular markers for pharmaceutical or diagnostics applications. Funding: The Academy of Finland, Systems Biology research programme, and the Graduate School in Computational Biology, Bioinformatics, and Biometry (ComBi). Collaborators: Riitta Lahesmaa (Turku Centre for Biotechnology), Tuula Nyman (University of Helsinki), Matej Orešic (VTT Biotechnology), Benno Schwikowski (Pasteur Institute, Paris), Mats Gyllenberg (University of Helsinki), Esa Uusipaikka (University of Turku), Samuel Kaski (Helsinki University of Technology), Timo Koski (Royal Institute of Technology, Stockholm), Eija Korpelainen (CSC – IT Center for Science), Jan Westerholm (Åbo Akademi University), Esa Tyystjärvi (University of Turku), and Mauno Vihinen (University of Tampere). Selected Publications: Okser, S., Lehtimäki, T., Elo, L.L., Mononen, N., Peltonen, N., Kähönen, M., Juonala, M., Fan, Y.M., Hernesniemi, J.A., Laitinen, T., Lyytikäinen, L.P., Rontu, R., Eklund, C., Hutri-Kähönen, N., Taittonen, L., Hurme, M., Viikari, J.S.A., Raitakari, O.T., and Aittokallio, T. (2010).Genetic variants and their interactions in the prediction of increased pre-clinical carotid atherosclerosis -- The Cardiovascular Risk in Young Finns Study, PLoS Genetics (in press). Eronen, V.P., Lindén, R.O., Lindroos, A., Kanerva, M., and Aittokallio T. (2010) Genome-wide scoring of positive and negative epistasis through decomposition of quantitative genetic interaction fitness matrices, PLoS ONE (in press). Moulder, R., Lönnberg, T., Elo, L.L., Filén, J.J., Rainio, E., Corthals, G., Orešic, M., Nyman, T.A., Aittokallio, T., and Lahesmaa, R. (2010) Quantitative proteomics analysis of the nuclear fraction of human CD4+ cells in the early phases of IL-4 induced Th2 differentiation, Molecular & Cellular Proteomics (in press). Lahti, L., Elo, L.L., Aittokallio, T., and Kaski, S. (2010) Probabilistic analysis of probe reliability in differential gene expression studies with short oligonucleotide arrays, IEEE Transactions on Computational Biology and Bioinformatics (in press). Codrea, M.C., Hakala-Yatkin, M., Kårlund-Marttila, M., Nedbal, L., Aittokallio, T., Nevalainen, O.S., and Tyystjärvi, E. (2010) Mahalanobis distance screening of Arabidopsis mutants with chlorophyll fluorescence, Photosynthesis Research (in press). 26 Elo, L.L., Järvenpää, H., Tuomela, S., Raghav, S., Ahlfors, H., Laurila, K., Gupta, B., Lund, R.J., Tahvanainen, J., Hawkins, R.D., Orešic, M., Lähdesmäki, H., Rasool, O., Rao, K.V.S., Aittokallio, T., and Lahesmaa, R. (2010) Genome-wide profiling of interleukin-4 and STAT6 transcription factor regulation of human Th2 cell programming, Immunity 32: 852-862. Elo, L.L., Mykkänen, J., Järvenpää, H., Nikula, T., Simell, S., Aittokallio, T., Hyöty, H., Ilonen, J., Veijola, J., Simell, T., Knip, M., Simell, O., and Lahesmaa, R. (2010) Early suppression of immune response pathways characterizes children with pre-diabetes in genome-wide gene expression profiling, Journal of Autoimmunity 35: 70-76. Aittokallio, T. (2010) Dealing with missing values in large-scale studies - microarray data imputation and beyond, Invited Review, Briefings in Bioinformatics 11: 253-264. Korolainen, M.A., Nyman, T.A., Aittokallio, T., and Pirttilä, T. (2010) An update on clinical proteomics in Alzheimer’s research, Journal of Neurochemistry 112: 1386-1414. Laajala, E., Aittokallio T., Lahesmaa, R. and Elo, L.L. (2009) Probelevel estimation improves the detection of differential splicing in Affymetrix exon array studies. Genome Biology 10: R77. Laajala, T.D., Raghav, S., Tuomela, S., Lahesmaa, R., Aittokallio, T. and Elo, L.L. (2009) A practical comparison of methods for detecting transcription factor binding sites in ChIP-seq experiments. BMC Genomics 10:618. Salmi, J., Nyman, T.A., Nevalainen, O.S. and Aittokallio, T. (2009) Filtering strategies for improving protein identification in highthroughput MS/MS studies. Proteomics 9: 848-860. 27 Elo, L.L., Hiissa, J., Tuimala, J., Kallio, A., Korpelainen, E. and Aittokallio, T. (2009) Optimized detection of differential expression in global profiling experiments: case studies in clinical transcriptomic and quantitative proteomic datasets. Briefings in Bioinformatics 10: 547-555. Hiissa, J., Elo, L.L., Huhtinen, K., Perheentupa, A., Poutanen, M. and Aittokallio, T. (2009) Resampling reveals sample-level differential expression in clinical genome-wide studies. OMICS Journal of Integrative Biology 13: 381-396. Huhtinen, K., Suvitie, P., Hiissa, J., Junnila, J., Huvila, J., Kujari, H., Setälä, M., Härkki, P., Jalkanen, J., Fraser, J., Mäkinen, J., Auranen, A., Poutanen, M. and Perheentupa, A. (2009) Serum HE4 concentration differentiates malignant ovarian tumours from ovarian endometriotic cysts. Br J Cancer 100:1315-1319. Clément-Ziza, M., Malabat, C., Weber, C., Moszer, I., Aittokallio, T., Letondal, C. and Rousseau, S. (2009) Genoscape: a Cytoscape plug-in to automate the retrieval and integration of gene expression data and molecular networks. Bioinformatics 25: 2617-2618. Aittokallio, T. (2009) Module finding approaches for protein interaction networks. In: Li, X.-L. and Ng, S.-K. (eds.) Biological Data Mining in Protein Interaction Networks, Medical Information Science Series, Chapter 18, pp. 335-353. IGI Global, Hershey, Pennsylvania, U.S.A. Merisaari, H., Parkkola, R., Alhoniemi, E., Teräs, M., Lehtonen, L., Haataja, L., Lapinleimu, H., Nevalainen, O.S. (2009) Gaussian mixture model-based segmentation of MR images taken from premature infant brains. J Neurosci Methods 182: 110-122. PROTEIN KINASE REGULATION OF BRAIN DEVELOPMENT AND DISEASE http://www.btk.fi/index.php?id=1240 Principal investigator: Eleanor Coffey, Ph.D., Academy of Finland Research Fellow, Turku Centre for Biotechnology, Åbo Akademi and Turku University, BioCity, Tykistokatu 6B, FI-20521 Turku, Finland. Tel. +358-2-3338605, Fax. +358-2-3338000. Email: [email protected] Biography: Eleanor Coffey (b. 1967) graduated from Trinity College Dublin in 1990 and received her Ph.D. from the University of Dundee in 1994. She was awarded a Wellcome Trust fellowship to carry out postdoctoral research in Prof. Karl Åkerman’s laboratory from 1994-1997. In 1997 she founded the Neuronal Signaling group at Åbo Akademi and in 2000 joined Turku Centre for Biotechnology as a group leader in molecular and cellular biology. In addition to running a research group, she directs the Cell Imaging Core at Turku Centre for Biotechnology and coordinates the Nordic Network on Imaging in Biology and Medicine. She currently holds an Academy of Finland Research Fellow position. Personnel: Postdoctoral researcher: Minna Tuittila, Ph.D., Graduate students: Artur Padzik, M.Sc., Justyna Zdrojewska, M.Sc., Emilia Komulainen, M.Sc., Raghavendra Mysore, M.Sc., Yubao Wang, M.Sc., Undergraduate and exchange students: Hanna Heikelä, Lihua Sun, Agnieszka Bialek, Prasannakumar Deshpande. Description of the project: Neurodegenerative disorders such as Alzheimer’s and Parkinson’s disease as well as stroke are characterised by the irreversible loss of nerve cell function. These diseases for which no cure is known are among the most costly to society. The protein kinase JNK is recognised as a critical player in stroke and neurodegeneration. However exactly how this family of kinases mediates cell death in the brain remains largely unknown. Although targeting of JNK for drug-based therapy is already underway, our understanding of the physiological function of JNK in the brain is in its infancy. From left to right: Johannes Tuikkala, Heidi Vähämaa, Laura Elo, Rolf Linden, Jussi Salmi, Olli Nevalainen, Ville Koskinen, and Tero Aittokallio. 28 A major challenge for signal transduction therapy is to selectively target the pathological function of signaling molecules without interfering with important physiological roles. To achieve this, our lab established a proteomics-based screen to identify protein kinase substrates and thereby broaden our understanding of kinase function. While we have used this methodology to successfully identify both novel and known substrates for JNK, p38 and PIM kinases (collaboration with Päivi Koskinen), among others (collaboration Erwin Wagner), the main focus of our research is to elucidate the molecular mechanism of JNK and JNK targets in the brain. Identification of novel JNK targets such as SCG10 and MAP2, as well as other targets under study, has highlighted a critical role for JNK in maintaining microtubule homeostasis and subsequently regulating axodendritic architecture. Identification of the JNK phosphorylation site on kinesin-1 helped characterize a role for JNK in regulation of fast axonal transport in neurons. We combine biochemical, proteomic, cell biology and imaging methods 29 with neuronal and organotypic cultures as well as transgenic mice to validate kinase targets and elucidate their function. In collaboration with Laurent Nyguen, we have established methods to track radial migration of neurons in the developing telencephalon using 4D imaging. In addition, we are examining dendrite and spine morphology in JNK1-/- brains using lucifer yellow iontophoretic loading followed by quantitative 3D image analysis. An important finding that we stumbled upon regarding JNK function in the nervous system, was the compartmental segregation of physiological verses pathological JNK function to the cytoplasm and nucleus respectively. By using compartment-targeted peptide inhibitors of JNK, we have shown that nuclear JNK activity is critical for neuronal death in response to trophic deprivation (the type of neuronal death that occurs during brain development) and in response to excitotoxic stimuli (the type of neuronal death that occurs in epilepsy, stroke and contributes to neurodegenerative disorders). Interestingly, although JNK is highly localised to the cytoplasm in neurons, we have shown that cytosolic JNK does not to contribute to these modes of neuronal death. Instead, we find that cytosolic JNK regulates physiological processes that maintain neuritic architecture and regulate migration. These functions of JNK are in turn mediated via cytosol-localised targets, independent of JNK-dependent transcriptional regulation. To realise the therapeutic potential of compartmental targeted JNK inhibitors, we are collaborating with Peter Clarke (University of Lausanne). This study investigates the value of nuclear-targeted peptide inhibitors of JNK as protectants from brain damage that occurs following stroke. By gaining information on how apoptotic and physiological functions of signaling molecules are partitioned within the cell allows selective targeting of inhibitors towards loci where pro-apoptotic events take place. This work provides proof of principal that subcellular targeting of inhibitor molecules provides increased specificity with reduced physiological disturbance. Funding: EU 6th framework STREP “STRESSPROTECT”, EU 6th framework ToK grant, “GAMIDI”, the Academy of Finland, Åbo Akademi University, Turku University Biomedical Sciences Graduate School, Finnish Graduate School in Neurosciences, Drug Discovery Graduate School, Magnus Ehrnrooth’s Stiftelse, CIMO, Sitra and the Torr Joe och Pentti Borgs Foundation. Collaborators: Michael Courtney (University of Kuopio), Tuula Kallunki (Danish Cancer Society), Thomas Herdegen (University of Kiel), Peter Clarke (University of Lausanne), Erwin Wagner (Research Institute of Molecular Pathology), Scott Brady (Univeristy of Illinois at Chicago), Laurent Nguyen (University of Liege), Päivi Koskinen (University of Turku), Aideen Long (Trinity College, Dublin). Selected Publications: Neurochem Int. J Immunol. 30 Mol Cell Biol. Nat Neurosci. Cellular Signalling, Cell Adhesion and Migration, Journal of Biological Chemistry, Molecular and Cellular Biology, Dan Johansen, L., Naumanen, T., Knudsen, A., Westerlund, N., Gromova, I., Junttila, M., Nielsen, C., Bottzauw, T., Tolkovsky, A., Westermarck, J., Coffey, E.T., Jäättelä, M., Kallunki, T. (2008) IKAP localizes to membrane ruffles with filamin A and regulates actin cytoskeleton organization and cell migration. Journal of Cell Science, 121:854-64. Westerlund, N., Zdrojewska, J., Courtney, M., Coffey, E. (2008) SCG10 as a molecular effector of JNK1: Implications for the therapeutic targeting of JNK in nerve regeneration. Expert Opinion on Therapeutic Targets. 12:31-43. Review. Semanova, M.M., Mäki-Hokkanen, A.M.J., Cao, C., Komarovski, V., Forsberg, K.M., Koistinaho, M., Coffey, E.T., Courtney, M.J. (2007) Rho mediates calcium-dependent activation of p38a and subsequent excitotoxic cell death. Nature Neuroscience, 10(4):436443. Tararuk, R., Östman, N., Li, W., Björkblom, B., Padzik, A., Zdrojewska, J., Hongisto, V., Herdegen, T., Konopka, W., Courtney, M.J., Coffey, E.T. (2006) JNK1 phosphorylation of SCG10 determines microtubule dynamics and axodendritic length. Journal of Cell Biology. 173: 265-277. Björkblom, B., Östman, N., Hongisto, V., Komarovski, V., Filen, J., Nyman, T., Kallunki, T., Courtney, M., Coffey, E. (2005) Constitutively active cytoplasmic JNK1 is a dominant regulator of dendritic architecture; role of MAP2 as an effector. Journal of Neuroscience. 25: 6350-6361. Yang, J., Lindahl, M., Lindholm, P., Virtanen, H., Coffey, E., Runeberg-Roos, P., Saarma, M. (2004) PSPN/GFRalpha4 has a significantly weaker capacity than GDNF/GFRalpha1 to recruit RET to rafts, but promotes neuronal survival and neurite outgrowth. FEBS Letters.569: 267-271. 31 Cao, J., Semenova, M.M., Solovyan, V.T., Han, J., Coffey, E.T., Courtney, M.J. (2004) Distinct requirements for p38alpha and c-Jun N-terminal kinase stress-activated protein kinase s in different forms of apoptotic neuronal death. Journal of Biological Chemistry. 279: 35903-35913. Hongisto, V., Smeds, N., Brecht, S., Herdegen, T., Courtney, M.J., Coffey, E.T. (2003) Lithium blocks the c-Jun stress response and protects neurons via its action on glycogen synthase kinase 3. Molecular and Cellular Biology. 23: 6027-6036. Coffey, E.T., Smiciene, G., Hongisto, V., Cao, J., Brecht, S., Herdegen, T., Courtney, M.J. (2002) c-Jun N-terminal protein kinase (JNK) 2/3 is specifically activated by stress, mediating c-Jun activation, in the presence of constitutive JNK1 activity in cerebellar neurons. Journal of Neuroscience. 22: 4335-4345. Hietakangas, V., Elo, I., Rosenstrom, H., Coffey, E.T., Kyriakis, J.M., Eriksson, J.E., Sistonen, L. (2001) Activation of the MKK4-JNK pathway during erythroid differentiation of K562 cells is inhibited by the heat shock factor 2-beta isoform. FEBS Letters. 505: 168-172. Coffey, E.T., Hongisto, V., Dickens, M., Davis, R.J. and Courtney, M.J. (2000) Dual roles for c-Jun N-terminal kinase in developmental and stress responses in cerebellar granule neurons. Journal of Neuroscience. 20: 7602-7613. Courtney, M.J. and Coffey, E.T. (1999) The mechanisms of ARA-C induced apoptosis of differentiating cerebellar granule neurons. European Journal of Neuroscience. 11: 1073-1084. Biochem. Soc. Trans. Courtney, M.J., Åkerman, K.E.O. and Coffey, E.T. (1997) Neurotrophins protect cultured cerebellar granule cells against the early phase of cell death by a two-component mechanism. Journal of Neuroscience. 17: 4201-4211. From left to right, front row: Hasan Mohammad, Emilia Komulainen, Justyna Zdrojewska, Eleanor Coffey, back row: Raghavendra Mysore, Lihua Sun, Prasannakumar Deshpande. Missing from the photo: Artur Padzik, Hanna Heikelä. 32 Organisation of Neuronal Signaling Pathways Principal investigator: Michael Courtney, Ph.D., Research manager, Professor of Cell Signaling. Contact information: Molecular Signaling Laboratory, Department of Neurobiology, A.I. Virtanen Institute, University of Kuopio, P.O. Box 1627, Neulaniementie 2, FIN-70211 Kuopio, Finland. Email: [email protected] Biography: Michael Courtney (b. 1967) graduated from University of Cambridge in 1988 (B.A.), and the University of Dundee in 1991(Ph.D). Postdoctoral fellowships from the Royal Society, Wellcome Trust, Academy of Finland and Sigrid Jusélius Foundation supported his quantitative imaging development and application activities from 1992 in Prof. Karl Åkerman’s laboratory in Åbo Akademi, Turku. After group leader positions at BTK from 1998, he was appointed from 2000 to a position at the A.I. Virtanen Institute, Kuopio and from 2006 to BTK. He has been affiliated with the Cell Imaging Core since its inception, and established and is running the Multimodal Imaging Unit at Kuopio University. He was appointed to an Academy of Finland Researcher post from 2003-2008, and Professor of Cell Signaling at the University of Kuopio from 2008. Personnel: Post-doctoral researchers: Franz Ho Ph.D.Ph.D., Peter Martinsson Ph.D., Minna Tuittila Ph.D., Olga Vergun Ph.D. Graduate students: Dorota Kaminska M.Sc., Kaisa Kosonen B.Sc., Lili Li, B.Sc., Xiaonan Liu, M.Sc., Maykel Lopez-Rodriguez M.Sc., Leena Yadav M.Sc. Undergraduate students: Tim Church, Soila Tossavainen Description of the project: Neuronal cells possess a complex architecture consisting of multiple subcellular compartments. Disease states place cells under stressful conditions. The p38 and JNK stress-activated protein kinase pathways are widely accepted to play a significant role in cell death in and outside the nervous system, and drugs directly targeting stress activated protein kinases have been under development for many years. However, these pathways also contribute to development, differentiation, and even survival and proliferation. This suggests that direct stress-activated protein kinase inhibitors may be of only limited use. In order to exploit the pathways for the development of novel neuroprotective drugs, it will be necessary to elucidate the mechanisms that organise these pathways into pools with neurodegenerative or physiological functions within the complex structure of neuronal cells. Only then can the neurodegenerative activities of the pathways be selectively eliminated. It has been suggested that this may help reduce the neuronal death that contributes to neurodegenerative conditions such as Alzheimer’s and Parkinson’s diseases, increasingly major causes of death, disability and socioeconomic impact in society Previous studies of mammalian stress-activated MAPK pathway have revealed the existence of a plethora of upstream regulators competent to recruit this pathway. In particular, proteins with putative scaffolding actions have been found. Such components could in principle have a number of effects on the associated upstream regulator, including (i) to potentiate their ability to activate 33 the pathway, (ii) to restrict accessibility to activators, (iii) to channel the downstream consequence to select targets and (iv) to localise these properties to specific compartments within a cell. Our lab’s aim is to elucidate how neuronal cells compartmentalise the endogenous components of the stress-activated protein kinase pathway and how specific stresses recruit only select components of these pathways. To achieve this, we currently focus on 3 areas: i) The impact of post-synaptic density proteins on neuronal stressactivated protein kinase signaling pathways; ii) Small G-protein signaling pathways regulating stress-activated protein kinases in neurons; iii) Development and implementation of approaches to imaging of intracellular signaling pathways. Recent publications revealed protein complexes of the postsynaptic compartment as upstream regulators of p38MAPK in neuronal stress responses (Cao et al., 2005; Semenova et al., 2007). The mechanisms which maintain selective responsiveness to upstream stimuli and restricted downstream consequences are anticipated to be a fruitful source of potential targets for future neuroprotective strategies. Thus we also utilise the information gleaned from studies of neuronal signaling mechanisms to develop and evaluate novel neuroprotective molecules in cooperation with collaborating partners from both the pharmaceutical industry and from academia. While pursuing these scientific goals, we also implement imaging methodologies. We adapt and establish the use of a wide range of FRET-based probes of cell signaling and on multiparameter imaging that allows spatiotemporal measurement of several pathways simultaneously in the same cells. We established facilities (physically located within Biocentre Kuopio, www.uef.fi/aivi/muic) to make available to all researchers live cell high High-Content Analysis (HCA)), as well as TIR-FRET and TIR-FRAP techniques. Total Internal Reflection methods exploit the spatially restricted evanescent wave formed at the interface between media of different refractive indices, thereby surpassing the classical diffraction limits. These methods are ideally suited to measure signaling events and protein turnover at protein complexes in the plasma-membrane proximal zones of living cells. The live-cell HCA unit is nationally unique and is now supported as a platform by two Biocenter Finland networks as part of the national Biocenter infrastructure. Funding: The Academy of Finland, The EU 6th framework STREP “STRESSPROTECT”, the EU 7th framework project “MEMOLOAD”, The Sigrid Juselius Foundation, The University of Kuopio, The Drug Discovery Graduate School, The Molecular Medicine Graduate School. Collaborators: Eleanor Coffey and Tassos Papageorgiou (BTK, Åbo Akademi and University of Turku), Christophe Bonny (University of Lausanne and Xigen Pharma AG), Denise Manahan-Vaughan (University of Bochum), Mark Spaller (Brown University, Providence, RI), Olli Pentikäinen (University of Jyväskylä), Antti Poso (University of Eastern Finland), Markus Rehm (Royal College Of Surgeons of Ireland), Anita Truttman (CHUV, Lausanne University Hospital), Mingjie Zhang (Hong Kong Institute of Science and Technology). 34 Selected Publications: Waetzig, V., Wacker, U., Haeusgen, W., Björkblom, B., Courtney, M.J., Coffey, E.T. and Herdegen, T. (2009) Concurrent protective and destructive signaling of JNK2 in neuroblastoma cells. Cell Signal. 21, 873-80 Hellwig, C.T., Kohler, B.F., Lehtivarjo A.-K., Dussmann, H., Courtney, M.J., Prehn, J.H. and Rehm, M. (2008) Real-time analysis of TRAIL/ CHX-induced caspase activities during apoptosis initiation. J. Biol. Chem. 283, 21676-85. Björkblom, B., Vainio, J.C., Hongisto, V., Herdegen, T., Courtney, M.J. and Coffey, E.T. (2008) All JNKs can kill but nuclear localization is critical for neuronal death. J. Biol. Chem. 283, 19704-19713. Hongisto, V., Vainio, J.C., Thompson, R., Courtney, M.J. and Coffey, E.T. (2008) The Wnt pool of GSK-3β is critical for trophic deprivation induced neuronal death. Mol. Cell. Biol. 28, 1515-1527. Westerlund, N., Zdrojewska, J., Courtney, M.J. and Coffey, E.T. (2008) SCG10 as a molecular effector of JNK1: Implications for the therapeutic targeting of JNK in nerve regeneration. Expert Opin. Ther. Targets, 12, 1-13. Semenova, M.M., Mäki-Hokkonen, A.M.J., Cao, J., Komarovski, V., Forsberg, K.M., Koistinaho, M. Coffey E.T. and Courtney, M.J. (2007) Rho mediates calcium-dependent activation of p38α and subsequent excitotoxic cell death. Nat. Neurosci. 10, 436-443. Tararuk, T., Östman N., Li, W., Björkblom, B., Padzik, A., Zdrojewska, J., Hongisto, V., Herdegen, T., Konopka, W., Courtney M.J. and Coffey, E.T. (2006) JNK1 phosphorylation of SCG10 determines microtubule dynamics and axodendritic length. J. Cell Biol. 173, 265-277. Björkblom, B., Östman, N., Hongisto, V., Komarovski, V., Filén, J., Nyman, T.A., Kallunki, T., Courtney, M.J. and Coffey, E.T. (2005) Constitutively active cytoplasmic JNK1 is a dominant regulator of dendritic architecture; role of MAP2 as an effector. J. Neurosci. 25, 6350-6361. Cao, J., Viholainen, J.I., Dart, C., Warwick, H.K., Leyland, M.L. and Courtney, M.J. (2005) The nNOS-PSD95 interface - a target for inhibition of excitotoxic p38 stress-activated protein kinase activation and cell death. J. Cell Biol. 168, 117-126. Cao, J., Semenova, M.M., Solovyan, V.T., Han, J., Coffey, E.T and Courtney, M.J. (2004) Distinct requirements for p38α and JNK stress-activated protein kinases in different forms of apoptotic neuronal death. J. Biol. Chem. 279, 35903-35913. Solovyan, V.T., Bezvenyuk, Z., Salminen, A., Austin, C.A. and Courtney M.J. (2002) The role of topoisomerase II beta in the excision of DNA loop domains during apoptosis. J. Biol. Chem. 277, 21458-21467. Coffey, E.T., Smiciene, G., Hongisto, V., Cao, J., Brecht, S., Herdegen, T. and Courtney, M.J. 2002) JNK2/3 is specifically activated by stress, mediating c-Jun activation, in the presence of constitutive JNK1 activity in cerebellar neurons. J. Neurosci. 22, 4335-4345. 35 Coffey, E.T., Hongisto, V., Davis, R.J., Dickens, M. and Courtney, M.J. (2000) Dual Roles for c-Jun N-terminal kinase in developmental and stress responses in cerebellar granule neurons. J. Neurosci. 20, 7602-7613. Courtney, M.J., Åkerman, K.E.O. and Coffey, E.T. (1997) Neurotrophins protect cultured cerebellar granule neurons against the early phase of cell death by a two-component mechanism. J. Neurosci. 17, 4201-4211. Courtney, M.J., Lambert, J.J. and Nicholls, D.G. (1990) The interactions between plasma membrane depolarization and glutamate receptor activation in the regulation of cytoplasmic free calcium in cultured cerebellar granule cells. J. Neurosci. 10, 38733879. Cytoskeletal and survival signaling Principal Investigator: John E. Eriksson, Ph.D., Professor. Address: Dept. of Biology, Åbo Akademi University, FI-20520 Turku, Finland. Laboratory address: Turku Centre for Biotechnology, BioCity, Tykistökatu 6B, P.O. Box 123, FIN-20521 Turku, Finland. Tel. int. + 358–2–333 8036, fax int. +358–2–3338000. E-mail: [email protected] Biography: John E. Eriksson (b. 1957) received his Ph.D. at the Åbo Akademi University in 1990. He was a post-doctoral fellow at Northwestern University in Dr. Robert D. Goldman’s laboratory during 1990-1993 (Fogarty International Fellowship from the National Institutes of Health 1991-1993). In November 1993 he joined the Centre for Biotechnology as a senior research fellow in cell biology. In 1999 he was appointed as Professor of Zoology at the Department of Biology, University of Turku. In 2006 he was appointed as Professor of Cell Biology at the Department of Biology, Åbo Akademi University and became Head of Cell Biology at the department in 2007. Description of the Project: Reversible protein phosphorylation is a key determinant in many fundamental cellular functions, such as survival, differentiation, structural organization, and stress responses. We are especially interested in phosphorylation-mediated signaling that maintains normal cellular and structural homeostasis, and how disturbances in the processing of this type of signaling are reflected as alterations in cellular survival and organization. As model systems for signal processing, we are studying apoptotic, stress-mediated, and cytoskeletal signaling, and their interrelationship. By exploring the interactions between these completely different signaling modes, we hope to advance our understanding how critical intracellular signals are processed and integrated. We have observed that growth signaling through the mitogenactivated kinase (MAPK) pathway has a dominant inhibiting effect on apoptosis induced by death receptors (Fas, TRAIL, and TNF receptors). We have shown that this mode of regulation has ramifications both in regulating death receptor responses of recently activated T-cells and in the resistance of certain tumor cell lines to death receptor stimulation. We have determined that the MAPK-mediated inhibition takes place by inhibiting the apoptotic signaling from the death-inducing signaling complex (DISC) of proteins assembled at the death receptor. We are determining the molecular mechanisms of this inhibition and how the DISC proteins and their interactions are regulated. Apoptosis is also affected by stress-mediated signaling. We have observed that stress facilitates death receptor-mediated apoptosis in a independently of heat shock protein expression. The stress-mediated sensitization is due to selective degradation of FLIP, a specific inhibitor of death receptor signaling. Targeted FLIP degradation by ubiquitylation is also responsible for the sensitization to TRAIL receptor-induced apoptosis that we observed in differentiating erythroid cells. We have found a PKCalpha/beta-mediated signaling module that regulates the turnover FLIP by an isoform and phosphorylation site-specific mechanism. These findings help understanding the regulation of death receptor responses during stress, fever, or inflammation, as well as during differentiation-related processes. 36 37 Intermediate filaments (IFs) are major cytoskeletal proteins important for ultrastructural organization and protection against various mechanical and other types of stresses. Recent studies have provided evidence that IFs are also involved in regulation of signaling. We have demonstrated that reversible phosphorylation is the key mechanism behind the assembly dynamics of IF proteins. Moreover, we have established that intermediate filaments are important signaling determinants, a question that relates to how the organization of the cytoskeleton will affect different signaling modules. By employing the interactions of different IFs (keratin 8/18, vimentin, nestin) with their signaling partners as models, we have elucidate the relationship between the cytoskeletal structure and the signaling state of the cell, and how this relationship will affect cell differentiation, growth, and survival. We observed that IFs act as general scaffolds for signaling proteins, and have focused on the association of IFs with JNKs, Cdk5, PKC isoforms, 14-3-3, and surface adhesion molecules are all involved in key regulatory processes in the cell. Recently, we determined that vimentin is a regulator of lymphocyte adhesion and transcellular migration, showing that the vimentin IFs form a highly dynamic anchoring structure, which is involved in organizing the surface molecules crucial for the migration. Another topical highlight includes the discovery of nestin as regulator of Cdk5 signaling. We have shown that nestin forms a scaffold and rheostat for the Cdk5/p35 signaling complex and shown that this function is important both during the differentiation of muscle cells and in apoptosis of neuronal cells.. Collaborators: The studies on apoptosis-related signaling are done in collaboration with Birgit Lane and David Lane (Institute of Medical Biology, A*Star, Singapore) and Lea Sistonen (Turku Centre for Biotechnology). The studies on IF-related signaling functions are carried out as a collaboration with Robert Goldman (Northwestern Univ., Chicago, USA), Johanna Ivaska (Univ. of Turku), Sirpa Jalkanen (Univ. of Turku), Hannu Kalimo (Univ. of Turku), Andras Nagy (Univ. of Toronto, Canada) and Bishr Omary (Stanford University, Palo Alto, USA). Funding: The Academy of Finland, TEKES, the European Union, the Finnish Cancer Organizations, the Sigrid Jusélius Foundation, and the Åbo Akademi Foundation. Selected Publications: Rosenholm J.M., Peuhu E., Bate-Eya L.T., Eriksson J.E., Sahlgren C. & Lindén M. (2010). Cancer-cell-specific induction of apoptosis using mesoporous silica nanoparticles as drug-delivery vectors. Small 6:1234-1241. Blomster H.A., Imanishi S.Y., Siimes J., Kastu J., Morrice N.A., Eriksson J.E. & Sistonen L. (2010). In vivo identification of sumoylation sites by a signature tag and cysteine-targeted affinity purification. J. Biol. Chem. 285:19324-9 de Thonel A., Ferraris S.E., Pallari H.M., Imanishi S.Y., Kochin V., Hosokawa T., Hisanaga S., Sahlgren C. & Eriksson J.E. (2010). Protein kinase Czeta regulates Cdk5/p25 signaling during myogenesis. Mol. Biol. Cell 21:1423-1434. Shen W.J., Patel S., Eriksson J.E., Kraemer F.B. Vimentin is a functional partner of hormone sensitive lipase and facilitates lipolysis. J. Proteome Res. 9:1786-1794. 38 Peuhu E., Rivero-Müller A., Stykki H., Torvaldson E., Holmbom T., Eklund P., Unkila M., Sjöholm R. & Eriksson J.E. (2010). Inhibition of Akt signaling by the lignan matairesinol sensitizes prostate cancer cells to TRAIL-induced apoptosis. Oncogene 29:898-908. Imanishi S.Y., Kouvonen P., Smått J.H., Heikkilä M., Peuhu E., Mikhailov A., Ritala M., Lindén M., Corthals G.L. & Eriksson J.E. (2009). Phosphopeptide enrichment with stable spatial coordination on a titanium dioxide coated glass slide. Rapid Commun. Mass Spectrom. 23:3661-3667. Rosenholm J.M., Peuhu E., Eriksson J.E., Sahlgren C. & Lindén M. (2009). Targeted intracellular delivery of hydrophobic agents using mesoporous hybrid silica nanoparticles as carrier systems. Nano Lett. 9:3308-3311. Eriksson J.E., Dechat T., Grin B., Helfand B., Mendez M., Pallari H.M., Goldman R.D. (2009). Introducing intermediate filaments: from discovery to disease. J. Clin. Invest. 119:1763-1771 (review). Rosenholm J., Meinander A. Peuhu E., Niemi R., Eriksson J.E., Sahlgren C. & Lindén M. (2009). Selective uptake of porous silica nanoparticles by cancer cells. Amer. Chem. Soc. 27:197-206. Kaunisto A, Kochin V, Asaoka T, Mikhailov A, Poukkula M, Meinander A. & Eriksson JE. (2009). PKC-mediated phosphorylation regulates c-FLIP ubiquitylation and stability. Cell Death Differ.16:1215-26. Mikhailov A., Sokolovskaya A., Yegutkin G.G., Amdahl H., West A., Yagita H., Lahesmaa R., Thompson L.F., Jalkanen S., Blokhin D. & Eriksson J.E. (2008). CD73 participates in cellular multiresistance program and protects against TRAIL-induced apoptosis. J. Immunol. 181: 464-75. Meinander, A., Söderström, T.S., Kaunisto, A., Poukkula, M., Sistonen, L. and Eriksson, J.E. (2007) Fever-like hyperthermia controls T-lymphocyte persistence by inducing degradation of c-FLIPshort. J. Immunol. 178: 3944-53. Imanishi S.Y., Kochin V., Ferraris S.E., deThonel A., Pallari H-M., Corthals G.L. & Eriksson J.E. (2007). Reference-facilitated phosphoproteomics: fast and reliable phosphopeptide validation by mikro-LC-ESI-Q-TOF MS/MS. Mol. Cell. Proteomics 6: 13801391. Nieminen, M., Henttinen, T., Merinen, M., Marttila-Ichihara, F., Eriksson, J.E. and Jalkanen S. (2006) Vimentin function in lymphocyte adhesion and transcellular migration. Nat. Cell Biol. 8: 156-162. Kochin, V., Imanishi S.Y. and Eriksson, J.E. (2006) Fast track to a phosphoprotein sketch – MALDI-TOF characterization of TLCbased tryptic phosphopeptide maps at femtomolar detection sensitivity. Proteomics 6: 5676-82. Sahlgren, C.M., Pallari, H-P., He, T., Chou, Y-H., Goldman, R.D. and Eriksson, J.E. (2006) An essential role of a nestin scaffold for regulation of Cdk5/p35 signaling in oxidant-induced death of neuronal progenitor cells. EMBO J 25: 4808-4819. 39 Imanishi, S.Y., Kochin, V. and Eriksson, J.E. (2006) Optimization of phosphopeptide elution conditions in immobilized Fe(III) affinity chromatography. Proteomics 7: 174-176. Pallari, H.M. and Eriksson, J.E. (2006) Intermediate filaments as signaling platforms. Science STKE. 19: pe53 (review). Söderström, T.S., Nyberg, S., Nieminen, M.I. and Eriksson, J.E. (2005) CD95 capping is ROCK-dependent and dispensable for apoptosis. J. Cell Sci. 118: 2211-2223. Poukkula, M., Kaunisto, A., Hietakangas, V., Denessiouk, K., Katajamäki, T., Johnson, M.J., Sistonen, L. and Eriksson, J.E. (2005) Rapid turnover of c-FLIPshort is determined by its unique C-terminal tail. J. Biol. Chem. 280: 27345-27355. Goswami, A., Burikhanov, R., de Thonel, A., Fujita, N., Goswami, M., Zhao, Y., Eriksson, J.E., Tsuruo, T. and Rangnekar, V.M. (2005). Binding and phosphorylation of Par-4 by Akt is essential for cancer cell survival. (2005) Mol. Cell. 20: 33-44. Eriksson, J.E., He, T., Trejo-Skalli, A.V., Härmälä-Brasken, A.S., Hellman, J., Chou, Y.H. and Goldman, R.D. (2004) Specific in vivo phosphorylation sites determine the assembly dynamics of vimentin intermediate filaments. J. Cell Sci. 117:919-32. Hietakangas, V., Poukkula, M., Heiskanen, K.M., Karvinen, J.T., Courtney, M.J., Sistonen, L. and Eriksson, J.E. (2003) Erythroid differentiation in K562 leukemia cells leads to sensitization to TRAIL-induced apoptosis by downregulation of FLIP. Mol. Cell. Biol. 23: 1278-1291. Hietakangas, V., Poukkula, M., Heiskanen, K.M., Karvinen, J.T., Sistonen, L. and Eriksson, J.E. (2003) Erythroid differentiation in K562 leukemia cells leads to sensitization to TRAIL-induced apoptosis by downregulation of FLIP. Mol. Cell. Biol. 23: 12781291. Sahlgren, C.M., Mikhailov, A., Vaittinen, S., Pallari, H.M., Kalimo, H., Pant, H.C. and Eriksson, J.E. (2003) Cdk5 regulates the organization of Nestin and its association with p35. Mol. Cell. Biol. 23:5090-5106. Tran, S.E.F., Meinander, A., Holmström, T.H., Rivero-Muller, A., Heiskanen, K.M., Linnau, E.K., Courtney, M.J., Mosser, D.D., Sistonen, L. and Eriksson, J.E. (2003) Heat stress downregulates FLIP and sensitizes to Fas receptor-mediated apoptosis. Cell Death Differ. 10: 1137-1147. Tran, S.E.F., Holmström, T.H., Ahonen, M., Kähäri, V-M. and Eriksson J.E. (2001) MAPK/ERK overrides the apoptotic signaling from Fas, TNF, and TRAIL receptors. J. Biol. Chem. 276: 1648416490. Holmström, T.H., Schmitz, I., Söderström, T., Poukkula, M., Johnson, V.L., Krammer, P. H., Chow, S.C. and Eriksson, J.E. (2000) MAPK/ERK signaling in activated T cells inhibits CD95/Fasmediated apoptosis downstream of DISC assembly. EMBO J. 19: 5418-5428. 40 From left to right, front row: Jenny Niinimäki, Petra Sonneborn, Emilia Peuhu, Tomoko Asaoka, Gloriane Lazaro, Susumu Imanishi, second row: Hanna-Mari Pallari, Kimmo Isoniemi, Juha Kastu, Claire Hyder, Erik Niemelä, Jenny Niinimäki, Vitaly Kochin, John Eriksson. 41 Cell Adhesion and Cancer http://www.btk.fi/index.php?id=1816 Principal investigator: Professor Johanna Ivaska, Ph.D.Ph.D., VTT Medical Biotechnology, Itäinen Pitkäkatu 4C, FI-20520 Turku, Finland; Phone: + 358 20 722 2807; FAX: + 358 20 722 2840, email: [email protected] Biography: Johanna Ivaska (b. 1972) received her MSc in Biochemistry in 1995 and Ph.D. in 2000 from the University of Turku. In 2000 she received a Post-doctoral Fellowship from the Academy of Finland. In 2001 she received the EMBO Long Term Fellowship. She was a post-doctoral fellow at Cancer Research UK in Prof. Peter Parker’s laboratory during 2000-2003. She returned to Finland in 2003 and joined VTT Medical Biotechnology and University of Turku Centre for Biotechnology as senior research fellow of the Academy of Finland and established her own research group. She was selected as a member of the EMBO Young Investigator program for 2007-2009. She was nominated professor of Molecular Cell Biology at University of Turku for 2008-2014 and her research group received ERC Starting Grant funding for 2008-2012 in their Cancer Signalosome project. Personnel: Post-docs: Elina Mattila, Ph.D.; Jeroe Pouwels, Ph.D.; Stefan Veltel, Ph.D.; Ghaffar Muharram, Ph.D.. Graduate students: Jonna Nevo, MSc; Karoliina Vuoriluoto, MSc; Anja Mai, MSc; Saara Tuomi, MSc; Antti Arjonen, MSc; Reetta Virtakoivu, MSc; Gunilla Högnäs; MSc. Technicians: Heidi Jalonen, Jenni Siivonen (both on maternity leave. Substituted by Laura Lahtinen and Emilia Helminen). Description of the project We investigate the relationship between cell adhesion and cancer. Cancer is a disease where cells grow out of control and invade, erode and destroy normal tissue. Invasive and metastatic behaviour of malignant cells is the major cause of mortality in all cancer patients. Migration and cell proliferation are critically regulated by physical adhesion of cells to each other and to their non-cellular surroundings (i.e. extracellular matrix) mediated by a family of adhesion receptors called integrins. Adhesion dependency of signalingsignaling pathways is well established but incompletely understood. In normal cells permissive signalingsignaling from integrins are prerequisite for receptor tyrosine kinase (RTKs) induced proliferation. This regulation is lost upon transformation. In the past few years, we have performed genome-wide screens to identify integrin-binding intracellular proteins to gain novel insight into integrin signalingsignaling and traffic in cancer cells. Our results demonstrate that integrins can also convey negative regulation on RTKs via a mechanism that is often lost in epithelial carcinomas. Our aim is to extend our studies on identifying integrin binding proteins to understand the diverse and sometimes unexpected biological roles of integrins. In addition to defining cytoplasmic integrin triggered pathways, we propose to investigate spatially regulated integrin membrane complexes. We aim to understand adhesion regulated signaling and the biological function of integrin membrane traffic in human malignancies. 42 From left to right, sitting: Ghaffar Muharram, Johanna Ivaska, Reetta Virtakoivu, second row: Jeroen Pouwels, Jarkko Heiskanen, Saara Tuomi, Elina Mattila, Stefan Veltel, Gunilla Högnäs, Laura Lahtinen, Jonna Nevo, Anja Mai, Emilia Helminen. Selected Publications: Tuomi, S., Mai, A., Nevo, J., Laine, JO, Vilkki, V., Öhman, TJ., Gahmberg, CG., Parker, PJ. and Ivaska, J. (2009) PKC Regulation of an 5 Integrin–ZO-1 Complex Controls Lamellae Formation in Migrating Cancer Cells. Science Signaling, 2 (77): ra32 Nevo, J., Mattila, E., Pellinen, T., Yamamoto, D.L., Sara, H., Iljin, K., Kallioniemi, O., Bono, P., Joensuu, H., Wärri, A. and Ivaska, J. (2009) Mammary Derived growth inhibitor facilitates escape from EGFR inhibitory therapy. Clin. Cancer Res. 15:6570-6578. Mattila E, Marttila H, Sahlberg N, Kohonen P, Tahtinen S, Halonen P, Perala M, Ivaska J. (2010) Inhibition of receptor tyrosine kinase signalling by small molecule agonist of T-cell protein tyrosine phosphatase. BMC Cancer. 10(1):7. Pellinen T., Tuomi, S., Arjonen, A., Wolf, M., Edgren, H., Meyer, H., Grosse, R., Kitzing, T., Rantala, JK., Kallioniemi O., Fässler, R., Kallio, M., and Ivaska, J. (2008) Integrin traffic regulated by Rab21 is necessary for cytokinesis. Dev. Cell, 15:371-385.). Mattila, E., Koskinen, K., Salmi, M. and Ivaska, J. (2008) Protein tyrosine phosphatase TCPTP controls VEGFR-2 signalling. J. Cell Sci. 121:3570-80. Vuoriluoto, K., Jokinen, J., Salmivirta, M. Heino, J. and Ivaska, J. (2008) Distinct syndecans function as integrin α2β1 co-receptors in 2D and 3D collagen. Exp. Cell Res. 314:3369-81. 43 Pellinen T, Arjonen A, Vuoriluoto K, Kallio K, Fransen JA, Ivaska J. Small GTPase Rab21 regulates cell adhesion and controls endosomal traffic of beta1-integrins. J. Cell Biol. 2006 173:767-80. Pellinen T. and Ivaska J. Integrin traffic, invited commentary (2006). J Cell. Sci. 119:3723-31. Mattila E., Pellinen, T., Nevo, J., Vuoriluoto, K. Arjonen, A. and Ivaska, J (2005) Negative regulation of EGFR signalling via integrin α1β1-mediated activation of protein tyrosine phosphatase TCPTP. Nat. Cell Biol. 7: 78-85. Ivaska, J., Vuoriluoto, K., Huovinen, T., Izawa, I., Inagiki, K., and Parker, PJ. (2005) PKCβ-mediated phosphorylation of vimentin controls integrin recycling and motility. EMBO J. 24:3834-3845. HYPOXIA IN CELL SURVIVAL Principal investigator: Panu Jaakkola, M.D., Ph.D., Senior fellow of The Academy of Finland. Address: Turku Centre for Biotechnology, Biocity, Tykistökatu 6B, P.O. Box 123, FIN-20521, Turku, Finland, Tel. +358 2 3338030, Fax. +358 2 3338000, E-mail: [email protected] Biography: Panu Jaakkola (b. 1965) received his M.D. in 1992 and Ph.D. in 1998 at the University of Turku. In 1999 he received a Junior Fellowship from the Academy of Finland. He was a postdoctoral fellow at the University of Oxford in Prof. Peter Ratcliffe’s laboratory during 1999-2001. He joined the Turku Centre for Biotechnology in the fall 2001. In 2002 he was appointed as a senior fellow of the Academy of Finland. Personnel: Post-doctoral scientist: Juha Pursiheimo, (Ph.D.) Graduate students: Terhi Jokilehto, (M.Sc.), Pekka Heikkinen, (M.Sc.), Heidi Högel, (M.Sc.), Krista Rantanen, (M.Sc.) Technicians: Taina Kalevo-Mattila Undergraduate students: Marika Nummela, Katri Piilonen, Siri Tähtinen Description of the project: Hypoxia (reduced O2 tension) is the main tissue damaging factor in several ischemic diseases. In contrast to the normal tissue, tumours use hypoxia as a growth-promoting factor. During ischemic assaults such as strokes, hypoxia activates apoptosis and leads to severe tissue damage. During cancer progression hypoxia causes inhibition of apoptosis and enhances tumour aggressiveness and metastasis. In keeping with this, it has been known for much of the past century that hypoxia causes resistance cancer treatments - both to chemotherapy and radiotherapy - and leads to poor prognosis. The aim of the project is to reveal mechanisms by which hypoxia regulates survival decisions in ischemic diseases and cancer progression. Our group has undertaken two major avenues to tackle the issue. The reduced oxygen is sensed by a family of enzymes called the HIF prolyl hydroxylases (PHD1-3). Under normoxia the hypoxia-inducible factor (HIF) is hydroxylated by PHDs at critical proline residues. This leads to ubiquitylation and proteosomal destruction of HIF. Under hypoxic conditions the hydroxylation ceases and HIF is stabilised. HIF then exerts its effects by activation of at least 80 genes. These have key functions in glucose homeostasis, angiogenesis, as well as cell survival and metastasis formation. Our studies have revealed novel and separate functions for two PHD isoforms (PHD2 and -3) in regulating cell growth, differentiation of cancer cells as well as regulation of apoptosis. Besides studying several aspects of molecular and cellular biology of the hydroxylases, we study the clinical importance of these factors. Transforming growth factor-β (TGF-β) is one of the bestcharacterised tumour growth regulating factors. It restricts the growth of early stage tumours, but at later stages of tumour progression cancer cells begin to exploit it as a malignancy, invasion and metastasis promoting cytokine. This paradox of TGF-β was originally described in skin cancer models over ten years ago and since then the paradox has been recapitulated in several other cancer models. Our group has recently identified a putative mechanism by which this may occur. We have found that hypoxia 44 45 is an environmental factor in tumours that can convert the TGF-β response into supporting tumorigenesis. Mechanistically, this involves hypoxic dephosphorylation of a TGF-β effector Smad3. Moreover, we have recently discovered that hypoxia converts Smad7, an inhibitor of the TGF-β signaling, from an inhibitor into a promoter of cell invasion. Funding: The Academy of Finland, Sigrid Juselius Foundation, Emil Aaltonen Foundation Collaborators: Peter Ratcliffe and Chris Pugh (Oxford University, UK), Eric Metzen (Luebeck University, Germany), Joachim Fandrey (Essen University, Germany), Reidar Grenman (Turku University), Veli-Matti Kähäri (Turku University), Heikki Minn (PET Centre, Turku University Hospital) Epstein, A.C.R., Gleadle, J.M., McNeill, L.A., Hewitson, K.S., O’Rourke, J., Mole, D.R., Mukherji, M., Metzen, E., Wilson, M.I., Dhanda, A., Tian, Y.-M., Masson, N., Hamilton, D.L., Jaakkola, P., Barstead, R., Hodgkin, J., Maxwell, P.H., Pugh, C.W., Schofield, C.J., Ratcliffe, P.J. C.elegans EGL-9 and mammalian homologues define a family of dioxygenases that regulate HIF through prolyl hydroxylation. (2001) Cell 107; 43-54. Pursiheimo, J., Taskén, K., Jalkanen, M. and Jaakkola, P. Involvement of Protein Kinase A in FGF-2 Activated Transcription. (2000) Proc. Natl. Acad. Sci. USA, 97(1): 168–173. Cockman, M.E., Masson, N, Mole, D.R., Jaakkola, P, Chang, G.W., Clifford, S.C, Maher, E.R, Pugh, C.W., Ratcliffe, P.J., Maxwell, P.H. Hypoxia inducible factor-alpha binding and ubiquitylation by the von hippel-lindau tumor suppressor protein. (2000) J. Biol. Chem. 275: 25733-25741. Selected Publications: Heikkinen P., Nummela M., Kähäri V.M. and Jaakkola P.M. (2010). Hypoxia converts Smad7 from tumor suppressor into tumor promoter. Cancer Res., In Press. Heikkinen P.T., Nummela M., Leivonen S.K., Westermarck J., Hill C.S., Kähäri V.-M., Jaakkola P.M. (2010). Hypoxia activated Smad3-specific dephosphorylation by PP2A. (2010). J Biol.Chem., 285(6):3740-9. Epub 2009 Dec 1. Jokilehto T., Högel H., Heikkinen, P., Rantanen K., Elenius, K., Sundström J., Jaakkola P.M. (2010). Retention of prolyl hydroxylase PHD2 in the cytoplasm prevents PHD2-induced anchorageindependent carcinoma cell growth. Exp. Cell Res. 316(7):116978. Epub 2010 Feb 12. Pursiheimo J., Rantanen K., Heikkinen P.T., Johansen T., Jaakkola P.M. (2009). Hypoxia-activated autophagy accelerates degradation of SQSTM1/p62. Oncogene, 28(3):334-344. Rantanen K., Pursiheimo J., Högel H., Himanen V., Metzen E., Jaakkola P.M. (2008) Prolyl Hydroxylase PHD3 Activates Oxygendependent Protein Aggregation. Mol Biol Cell 19(5): 2231-40. Jokilehto, T., Rantanen, K., Luukkaa, M., Grenman, R., Minn, H., Kronqvist, P., Jaakkola P.M. (2006). Overexpression and nuclear translocation of HIF prolyl hydroxylase PHD2 in head and neck squamous cell carcinoma associates with tumor aggressiveness. Clin Cancer Res 12(4):1080-1087 Marxsen, J. H., Stengel, P., Doege, K., Heikkinen, P., Jokilehto, T., Wagner, T., Jelkmann, W., Jaakkola, P., and Metzen, E. (2004) Hypoxia-inducible factor-1 (HIF-1) promotes its degradation by induction of HIF-alpha-prolyl-4-hydroxylases. Biochem J 381, 761767 Jaakkola, P., Mole, D. R., Tian, Y. M., Wilson, M.I., Gielbert, J., Gaskell, S.J., Kriegsheim, Av, Hebestreit, H.F., Mukherji, M., Schofield, C.J., Maxwell, P.H., Pugh, C.W., Ratcliffe, P.J. Targeting of HIF-alpha to the von Hippel-Lindau ubiquitylation complex by O2-regulated prolyl hydroxylation. (2001) Science 292; 468-72. 46 From left to right, front row: Panu Jaakkola, Krista Rantanen, Taina Kalevo-Mattila, second row: Heidi Högel, Maiju Nuutila, Terhi Jokilehto, Marika Nummela. 47 Bioenergy Group http://www.btk.fi/index.php?id=1816 Principal Investigator: Patrik R. Jones, Ph.D., Group Leader, Turku collegium for Science and Medicine, University of Turku, Centre for Biotechnology, Turku BioCity, Tykistökatu 6A, 5krs, 20520, Turku. Tel.:+358-2-333-8094. Email: [email protected]. Biography: Patrik Jones (b. 1968) completed his undergraduate degree in Agricultural Sciences (Oenology, Honours) at the University of Adelaide and obtained his Ph.D. (2001) from the University of Adelaide, Australia, and the Royal Veterinary and Agricultural University of Copenhagen, Denmark, on the topic of plant natural product metabolism. Before commencing his current position in Turku in 2008, he held a position as JSPS-funded post-doctoral fellow (2001-2002, plant natural product metabolism) at Chiba University, Japan; Research Chemist (2003-2004, wine chemistry and sensory perception) at the Australian Wine Research Institute in Adelaide, Australia; Research Director (2005-2008, microbial metabolic engineering and renewable fuel production) at Fujirebio Inc. (100% for-profit), Tokyo, Japan. Personnel: Undergraduate students: Sanna Peltonen (Bioinformatics), Linda Vuorijoki (Biochemistry) Graduate student: Fernando Guerrero (funded by Center of Excellence in Integrative Photosynthesis and Bioactive Compound Research at Systems Biology Level) Description of the project: To reduce greenhouse gas emissions and implement new energy-production strategies that replace fossil fuels, alternative fuel production methods are needed in the future. We target fundamental and applied topics with the aim of contributing towards the development of novel sustainable technologies for the production of engine-ready fuels. The laboratory currently has two main lines of research: We study biochemical pathways of plant hydrocarbon biosynthesis in order to generate components for metabolic engineering. Photobiological model organisms are engineered in order to (a) introduce biofuel-pathways that do not exist in nature and (b) to modify host metabolism to favor those pathways. Funding: Varsinais-Suomi Cultural Foundation, Emil Aaltonen Foundation Selected publications: Akhtar, M.K. and Jones, P.R. (2009) Construction of a synthetic YdbK-dependent pyruvate:H2 pathway in Escherichia coli BL21(DE3). Metabolic Engineering 11, 139-147. Veit, A., Akhtar, K.M., Mizutani, T., Jones P.R. (2008) Constructing and testing the thermodynamic limits of synthetic NAD(P)H:H2Pathways. Microbial Biotechnology 1, 382-394. Akhtar, K.M., and Jones P.R. (2008) Deletion of iscR stimulates recombinant clostridial Fe-Fe hydrogenase activity and H2accumulation in Escherichia coli BL21(DE3). Applied Microbiology and Biotechnology 78(5), 853-862. Akhtar, K.M., and Jones P.R. (2008) Constructing a synthetic hydFhydE-hydG-hydA operon for engineering biohydrogen production. Analytical Biochemistry 373(1), 170-172. Park, M.-O., Mizutani, T., Jones. P.R. (2007) Glyceraldehyde3-phosphate:ferredoxin oxidoreductase from Methanococcus maripaludis. Journal of Bacteriology 189(20), 7281-7289. Jones, P.R. (2008) Improving fermentative biomass-derived H2production by engineering microbial metabolism. International Journal of Hydrogen Energy 33, 5122-5130. (1) Fundamental understanding for improving fermentative and photobiological H2-production. H2-pathways depend heavily on O2-labile Fe-S clusters for electron transfer. We study Fe-S cluster metabolism, with a particular focus on the repair of partially damaged clusters, in order to enhance H2-pathways in general and in order to contribute towards the development of O2-tolerant H2prodction strategies. Both fermentative and photobiological H2-pathways will depend on the use of NAD(P)H as a major electron-carrier, although NAD(P) H is a thermodynamically unfavorable electron-donor for H2pathways. The aim is to contribute towards an understanding of the mechanisms that are involved in the regulation of NADP(H)metabolism. Both topics are studied with a combination of computational and experimental sciences, with a focus on both individual key-reactions and systems-level regulation of metabolism. (2) Synthetic biology - metabolic pathway construction for synthesis of novel hydrocarbon transport fuels. 48 From left to right. Fernando Guerrero, Sanna Peltonen, Linda Vuorijoki, Patrik Jones. 49 KINETOCHORE AND CANCER RESEARCH GROUP Principal investigator: Marko Kallio, Ph.D. Docent, Senior Research Scientist and Team Leader, VTT Medical Biotechnology, Itäinen Pitkäkatu 4C, FI-20521, Turku, Finland and Turku Centre for Biotechnology, BioCity, Tykistökatu 6B, FI-20521 Turku, Finland. Tel. +358-(0)2-4788614, Fax. +358(0)20-7222840. Email: Biography: Marko Kallio (b. 1967) graduated in Genetics from University of Turku in 1992 and received his Ph.D. degree from Department of Human Genetics at University of Turku 1996 with an honorary mention. 1996-1998 Dr. Kallio was in the laboratory of Prof. Gary Gorbsky (Univ. Virginia, USA) as Post-doctoral Fellow and 19982000 in the laboratories of Prof. John Eriksson and Prof. Lea Sistonen (Univ. Turku, Finland) as a Senior Post-doctoral Fellow. 2000-2003 Dr. Kallio worked as an Assistant Research Professor at University of Oklahoma HSC, USA. His research group received Marie Curie Excellence grant for 2004-2008. In early 2004 Dr. Kallio moved back to Finland and has since been a group leader at VTT Medical Biotechnology, a research institute affiliated with the University of Turku. Personnel: Post-doctoral researchers: Kimmo Jaakkola, M.D., Leena Laine, Ph.D., Elli Narvi, Ph.D., Christina Oetken-Lindholm, Ph.D., Sebastian Winsel, Ph.D. Graduate students: Anu Kukkonen-Macchi, M.Sc., Jenni MäkiJouppila, M.Sc., Anna-Leena Salmela, M.Sc., Mariaana Vuoriluoto, M.Sc. Technicians: Pauliina Toivonen Alumni: Tim Holmström, Ph.D., Jeroen Pouwels, Ph.D., Oana Sicora, Ph.D., Asta Varis, Ph.D., Chang-Dong Zhang, Ph.D. Description of the projects: The Kinetochore and Cancer Team investigates mechanisms of cell division in somatic cells and in meiotic systems. Understanding cell division errors may help to explain origin of genomic instability and is expected to identify novel therapeutic possibilities for treatment of cancer. We have performed a number of high-throughput screens (HTS) for small molecules, siRNAs and miRNAs with antimitotic activity. We are interested of conditions that suppress cell’s viability as a consequence of premature inactivation of the spindle assembly checkpoint, a conserved signaling pathway monitoring fidelity of mitosis. Finally, we have launched a project to explore the mechanisms of acquired resistance to microtubule-drugs, a growing clinical problem in the treatment of cancer. Resistance to mt-drugs has links to malfunction of tubulin and mitotic checkpoint proteins but these mechanisms are poorly understood. Errors during cell division may result in unequal distribution of DNA between the daughter cells. Gain or loss in the number of chromosomes of the genome is a known cause for miscarriages and birth defects in human, and a hallmark of cancer. In our research, 50 we have focused to study of the spindle assembly checkpoint (also known as the mitotic or the kinetochore checkpoint) that monitors interactions between the spindle microtubules and kinetochores, the microtubule binding platforms of chromosomes. If mistakes occur in the microtubule-kinetochore connections, the mitotic checkpoint becomes active and prevents separation of sister chromatids until errors in the chromosome alignment are corrected. Although the main principles of the spindle assembly checkpoint are well documented many molecular details remain to be explored. To this end, we are investigating the function of Chromosome Passenger Complex (CPC) and Ndc80 complex required for the spindle assembly checkpoint signaling and ordered progression of mitosis. We are especially interested of Aurora B, Incenp, and Hec1 proteins that facilitate normal microtubule-chromosome associations. The specific question we wish to answer is how members of CPC proteins interact with each other and with mitotic co-factors, and how they contribute to mitotic checkpoint signaling. The findings are expected to catalyze cancer drug discovery by identification of new possibilities for Hec1 and Aurora B inhibition. In another set of projects, we are focusing on characterization of the several anti-mitotic lead compounds, siRNAs and miRNAs that we discovered during our recent HTSs. In particular, we are working to validate the mechanism of action of three putative anti-Hec1 compounds that effectively perturb normal mitosis and trigger cancer cell killing in cell culture assays. Moreover, we aim to determine the mechanism of action of the mitotic checkpoint suppressing siRNAs and miRNAs using various cell-based and in vitro assays. Lastly, in a collaborative project with Prof. Kallioniemi we have identified gene copy number and gene expression alterations in parental lung and ovarian cancer cell lines and their microtubuledrug resistant variants. To directly link these genomic findings with a drug efflux pump independent mechanisms of action of microtubule-drug resistance, we are testing if re-expression of the genes lost from the drug-resistant variant cell lines will re-sensitize them to microtubule-drugs, and to silence the same genes alone or in combinations in the parental lines to determine if their lost drives development of microtubule-drug resistance. Other factors that we have recently discovered to affect the sensitivity to microtubuledrugs are loss of certain tubulin isoforms that leads to abnormal function of mitotic spindle and precocious inactivation of the mitotic checkpoint causing formation of multinucleated progeny cells. We expect to identify novel molecular mechanisms for the microtubuledrug insensitivity. These findings may have diagnostics value in the development of individually optimized therapeutic regimens for cancer patients and for design of new class of microtubule-drugs that selectively target taxane-resistant tumours. Funding: VTT Technical Research Centre of Finland, The Academy of Finland, Finnish Cancer Organisations, TuBS and DDGS Graduate Schools, Bayer Schering Pharma Collaborators: Gary Gorbsky (OMRF, Oklahoma USA), Todd Stukenberg (Univ. Virginia, USA), Olli Kallioniemi (FIMM), 51 Selected Publications: Salmela AL, Pouwels J, Varis A, Kukkonen AM, Toivonen P, Halonen PK, Perälä M, Kallioniemi O, Gorbsky GJ, Kallio MJ. (2009) Carcinogenesis, 30:1032-1040. Ahonen LJ, Kukkonen AM, Pouwels J, Bolton MA, Jingle CD, Stukenberg PT, Kallio MJ. (2009). segregation, overrides the spindle checkpoint, and perturbs microtubule dynamics in mitosis. Curr. Biol. 12, 900-905. Giodini A, Kallio MJ, Wall NR, Gorbsky GJ, Tognin S, Marchisio PC, Symons M, Altieri DC. (2002) Regulation of microtubule stability and mitotic progression by survivin. Cancer Res. 62, 2462-2467. Chromosoma, 118:71-84. Pellinen T, Tuomi S, Arjonen A, Wolf M, Edgren H, Meyer H, Grosse R, Kitzing T, Rantala JK, Kallioniemi O, Fässler R, Kallio M, Ivaska J. (2008). Dev Cell, 15:371-385. Vorozhko VV, Emanuele MJ, Kallio MJ, Stukenberg PT, Gorbsky GJ. (2008) Multiple mechanisms of chromosome movement mediated through the Ndc80 complex and dynein/dynactin. Chromosoma, 117:169-179. (2007) Shugoshin 1 plays a central role in kinetochore assembly and is required for kinetochore targeting of Plk1. Cell Cycle. 6, 1579-1585. Wang YY, Parvinen M, Toppari J, Kallio MJ. (2006) Inhibition of Aurora kinases perturbs chromosome alignment and spindle checkpoint signaling in rat spermatocytes. Exp Cell Res. 312, 3459-3470. Ahonen LJ, Kallio MJ, Daum JR, Bolton M, Manke IA, Yaffe MB, Stukenberg PT, Gorbsky GJ. (2005) Polo-like kinase 1 creates the tension-sensing 3F3/2 phosphoepitope and modulates the association of spindle-checkpoint proteins at kinetochores. Curr Biol. 15, 1078-1089. Beardmore VA, Ahonen LJ, Gorbsky GJ, Kallio MJ. (2004) Survivin dynamics increases at centromeres during G2/M phase transition and is regulated by microtubule-attachment and Aurora B activity. J Cell Sci. 117, 4033-4042. McCleland ML, Kallio MJ, Barrett-Wilt GA, Kestner CA, Shabanowitz J, Hunt DF, Gorbsky GJ, Stukenberg PT. (2004) The vertebrate Ndc80 complex contains functional homologs of Spc24 and Spc25 and is required to establish and maintain kinetochore-microtubule attachment. Curr Biol. 14, 131-137. McCleland ML, Gardner RD, Kallio MJ, Daum JR, Gorbsky GJ, Burke DJ, Stukenberg PT. (2003) The highly conserved Ndc80 complex is required for kinetochore assembly, chromosome congression, and spindle checkpoint activity. Genes Dev. 17, 101114. Kallio MJ, Beardmore VA, Weinstein J, Gorbsky GJ. (2002) Rapid microtubule-independent dynamics of Cdc20 at kinetochores and centrosomes in mammalian cells. J. Cell Biol. 158, 841-847. Kallio MJ, McCleland ML, Stukenberg PT, Gorbsky GJ. (2002) Inhibition of aurora B kinase blocks chromosome 52 From left to right, front row: Marko Kallio, second row: Jenni Mäki-Jouppila, Christina Oetken-Lindholm, third row: Anu Kukkonen-Macchi, back row: Sebastian Winsel. 53 CANCEROMICS RESEARCH PROGRAMME Principal Investigator: Olli Kallioniemi, M.D., Ph.D., Director, Institute for Molecular Medicine Finland (FIMM), University of Helsinki, Tukholmankatu 8, 00140 University of Helsinki, Finland. Director of the Academy of Finland Centre of Excellence on Translational Genome-scale Biology, Medical Biotechnology, VTT Technical Research Centre of Finland and University of Turku. Laboratory address: Medical Biotechnology, PharmaCity, Itäinen Pitkäkatu 4C, P.O. Box 106, FI-20521 Turku, Finland. Tel. +358-20-722 2800. Fax +358-20-722 2840. Email: [email protected]. Biography: Dr. Olli Kallioniemi (born 1960) received his M.D. in 1984 and Ph.D. in 1988 at the University of Tampere in Finland. Olli Kallioniemi held several positions in the US over a 10-year period, most recently (1995-2002) as the Head of Translational Genomics Section at the Cancer Genetics Branch, National Human Genome Research Institute, at NIH, Bethesda, Maryland. Since 2003, he has been Professor of Medical Biotechnology at the VTT Technical Research Centre of Finland with a joint appointment at the University of Turku. Academy of Finland Professorship in 2004-2007. In 2007, he was nominated as a director of the Institute for Molecular Medicine Finland (FIMM), a Nordic EMBL Partnership in Molecular Medicine. He continues to direct the ongoing projects in Turku until the end of 2011. He is an author of 250 publications and editor or member of the editorial board of six journals. Inventor of 15 issued patents, with a focus on technology development, such as Comparative Genomic Hybridization (CGH) in 1992, tissue microarrays in 1998 and cell-based RNAi microarrays in 2003. EACR young investigator award in 1994, Anders Jahre Prize in 1998, NIH Director’s lecture in 2000, Medal of the Swedish Medical Society in 2003, National Academy of Sciences (Finland) in 2005, EMBO Membership in 2006, and the Abbot-IFCC award in Molecular Diagnostics 2009. Personnel: Ph.D-students: Anna Aakula, M.Sc., Elmar Bucher, M.Sc., Mari Björkman, M.Sc., Santosh Gupta, M.Sc., Kirsi Ketola, M.Sc., Pekka Kohonen, M.Sc. Paula Vainio, M.Sc., Sirkku Pollari, M.Sc., Coordinator: Riina Plosila, M.Sc. Description of the Project: The overall purpose of this research program is to develop and apply high-throughput technologies to understand mechanisms of progression of breast and prostate cancers as well as to identify mechanisms of drug response.. The aims of the various research programs are to: 1. Apply cancer genomics to identify key genes and pathways in 1. breast and prostate cancer 2. Apply high-throughput RNA interference and chemical biology 2. to identify living cells, with particular attention towards cancerspecific vulnerabilities and steroid-dependent signaling and 3. Translate the molecular discoveries towards drug discovery, 3. clinical diagnostics and personalized medicine. 54 We are using advanced systems biology and chemical biology approaches to characterize the deregulation of cancer cell functions. The research is carried out in collaboration between the Institute for Molecular Medicine Finland (FIMM), the Medical Biotechnology Centre of the VTT Technical Research Centre of Finland and the Turku Centre for Biotechnology. Our group coordinates Academy of Finland Centre of Excellence in Translational Genome-Scale Cell Biology. We have developed biochip technologies, bioinformatics, systems biology, translational cancer research and drug development technologies, such as cell microarrays, protein lysate microarrays, in silico profiling of gene expression in clinical samples and many others. Collaborators: Tomi Mäkelä, Lauri Aaltonen, Jussi Taipale, Päivi Ojala, Sampsa Hautaniemi, Heli Nevanlionna, Heikki Joensuu, Kari Alitalo, Jonathan Knowles, Emmy Verschuren, Sergey Kuzneshov, Krister Wennerberg (FIMM and Biomedicum Helsinki), Antti Poso, Tapio Visakorpi, Jukka Westermarck and many others in other Universities in Finland. We have over 100 collaborators in current EU projects such as Epitron, Genica, APO-SYS, Prosper and Meta-Cancer (FP7). Funding: The Academy of Finland, Tekes, Finnish Cancer Organizations and Sigrid Juselius Foundation. Our biggest source of funding comes from the EU framework projects, including Epitron, Genica, APOSYS, Prosper and Meta-Cancer (FP7). Selected recent publications: Rantala JK, Edgren H, Lehtinen L, Wolf M, Kleivi K, Moen Vollan HK, Aaltola A-R, Laasola P, Kilpinen S, Saviranta P, Iljin K, Kallioniemi O. Integrative Functional Genomics Analysis of Sustained Polyploidy Phenotypes in Breast Cancer Cells Identifies an Oncogenic Profile Role for GINS21,2. Neoplasia, in press, 2010 Gupta S, Iljin K, Sara H, Mpindi JP, Mirtti T, Vainio P, Rantala J, Alanen K, Nees M, Kallioniemi O. FZD4 as a Mediator of ERG Oncogene-Induced WNT Signaling and Epithelial-to-Mesenchymal Transition in Human Prostate Cancer Cells. Cancer Res. 2010 Aug 16. McBride DJ, Orpana AK, Sotiriou C, Joensuu H, Stephens PJ, Mudie LJ, Hämäläinen E, Stebbings LA, Andersson LC, Flanagan AM, Durbecq V, Ignatiadis M, Kallioniemi O, Heckman CA, Alitalo K, Edgren H, Futreal PA, Stratton MR, Campbell PJ. Use of cancerspecific genomic rearrangements to quantify disease burden in plasma from patients with solid tumors. Genes Chromosomes Cancer. 2010 Aug 19. International Cancer Genome Consortium. International network of cancer genome projects. Nature, 464(7291):993-998, 2010. Härmä V, Virtanen J, Mäkelä R, Happonen A, Mpindi JP, Knuuttila M, Kohonen P, Lötjönen J, Kallioniemi O, Nees M. A comprehensive panel of three-dimensional models for studies of prostate cancer growth, invasion and drug responses. PLoS One, 5(5):e10431, 2010. Pollari S, Käkönen SM, Edgren H, Wolf M, Kohonen P, Sara H, Guise T, Nees M, Kallioniemi O. Enhanced serine production by 55 bone metastatic breast cancer cells stimulates osteoglastogenesis. Breast Cancer Res Treat. 2010 Mar 30. Nevo J, Mattila E, Pellinen T, Yamamoto DL, Sara H, Iljin K, Kallioniemi O, Bono P, Heikkilä P, Joensuu H, Wärri A, Ivaska J. MammaryDerived Growth Inhibitor Alters Traffic of EGFR and Induces a Novel Form of Cetuximab Resistance. Clin Cancer Res.,15(21):6570-81, 2009. Main H, Lee KL, Yang H, Haapa-Paananen S, Edgren H, Jin S, Sahlgren C, Kallioniemi O, Poellinger L, Lim B, Lendahl U. Interactions between Notch- and hypoxia-induced transcriptomes in embryonic stem cells. Exp Cell Res., 316(9):1610-1624, 2010. Leivonen SK, Mäkelä R, Ostling P, Kohonen P, Haapa-Paananen S, Kleivi K, Enerly E, Aakula A, Hellström K, Sahlberg N, Kristensen VN, Børresen-Dale AL, Saviranta P, Perälä M, Kallioniemi O. Protein lysate microarray analysis to identify microRNAs regulating estrogen receptor signaling in breast cancer cell lines. Oncogene, 28(44):3926-3936, 2009. Iljin K, Ketola K, Vainio P, Halonen P, Kohonen P, Fey V, Grafström RC, Perälä M, Kallioniemi O. High-throughput cell-based screening of 4910 known drugs and drug-like small molecules identifies disulfiram as an inhibitor of prostate cancer cell growth. Clin Cancer Res., 15(19):6070-6078, 2009. Côme C, Laine A, Chanrion M, Edgren H, Mattila E, Liu X, Jonkers J, Ivaska J, Isola J, Darbon JM, Kallioniemi O, Thézenas S, Westermarck J. CIP2A is associated with human breast cancer aggressivity. Clin Cancer Res., 15(16):5092-5100, 2009. Varjosalo M, Björklund M, Cheng F, Syvänen H, Kivioja T, Kilpinen S, Sun Z, Kallioniemi O, Stunnenberg HG, He, W-W, Ojala P, Taipale J. Application of Active and Kinase-Deficient Kinome Collection for Identification of Kinases Regulating Hedgehog Signaling. Cell, 133:537-548, 2008. Pellinen T, Tuomi S, Arjonen A, Wolf M, Edgren H, Meyer H, Rantala JK, Kallioniemi O, Fässler R, Kallio M and Ivaska J. Integrin traffic regulated by Rab21 is necessary for cytokinesis. Dev. Cell., 15(3):371-385, 2008. Kilpinen S, Autio R, Ojala K, Iljin K, Bucher E, Sara H, Pisto T, Saarela M, Skotheim R, Björkman M, Mpindi J. P., HaapaPaananen S, Vainio P, Edgren H, Wolf M, Astola J, Nees M, Hautaniemi S, Kallioniemi Olli. Systematic bioinformatic analysis of expression levels of 17,330 human genes across 9,783 samples from 175 types of healthy and pathological tissues. Genome Biol., 9(9):R139, 2008. Björkman M, Kristiina I, Halonen P, Henri S, Kaivanto E, Nees M, Kallioniemi Olli. Defining the molecular action of HDAC inhibitors and synergism with androgen deprivation in ERG-positive prostate cancer. Int J Cancer, 123 (12):2774-2781, 2008. Iljin K, Wolf M, Edgren H, Gupta S, Kilpinen S, Skotheim R, Peltola M, Smit F, Verhaegh G, Schalken J, Nees M, Kallioniemi O. 2006. TMPRSS2 fusions with oncogenic ETS factors in prostate cancer involve unbalanced genomic rearrangements and are associated with HDAC1 and epigenetic reprogramming. Cancer Res., 66:10242-10246, 2006. Edgren, H. & Kallioniemi, O. Integrated breast cancer genomics. Cancer Cell, 10:453-454, 2006. From left to right, sitting: Paula Vainio, Mari Björkman, Riina Plosila, Pekka Kohonen, standing: Kirsi Ketola, Anna Aakula, Santosh Gupta, Olli Kallioniemi, Sirkku Pollari, Elmar Bucher. 56 57 Signaling Pathways Regulated by Oncogenic Pim Kinases Principal Investigator: Päivi J. Koskinen, Ph.D., Senior Assistant, Adj. Prof. in Molecular and Cell Biology. Laboratory address: Turku Centre for Biotechnology, BioCity, Tykistökatu 6 B, P.O. Box 123, FIN-20521 Turku, Finland. Tel. + 358-2-3338044, fax + 358-2-3338000. E-mail: [email protected] Biography: Päivi Koskinen (b. 1961) received her Ph.D. at the University of Helsinki in 1992. During years 1993-1996 she worked as a postdoctoral fellow in Dr. Robert Eisenman´s laboratory at the Fred Hutchinson Cancer Research Center in Seattle, USA. In 1996 she joined the Turku Centre for Biotechnology as a group leader and a research fellow of the Academy of Finland. Since 2006 she has been employed by the Department of Biology, University of Turku. so efficiently co-operate with Myc family transcription factors in murine, and most likely also in human tumorigenesis. Even though Myc-overexpressing cells proliferate faster, they are more prone to apoptosis, so it is advantageous for them to co-overexpress also Pim kinases, which regulate the balance between anti- and proapoptotic factors and boost activities of transcription factors that are essential for production of cytokines and other survival factors. To further characterize the signaling pathways downstream of Pim kinases, we have collaborated with the group of Eleanor Coffey (CBT) and used phosphoproteomics to reveal novel substrates for Pim kinases. These proteins have recently been confirmed as true Pim substrates and their functional validation is underway using both overexpression and RNA interference-based approaches. In collaboration with the group of Garry Corthals (CBT), we have developed sensitivity of the methodology to identify phosphopeptides. In addition, we have been collaborating with two groups of chemists (Jari Yli-Kauhaluoma, Viikki Biocenter, Helsinki and Pascale Moreau, CNRS, France) to identify and validate Pim- Personnel: Postdoctoral researchers: Eeva Rainio, Ph.D. Graduate students: Jouko Sandholm, M.Sc., Riitta Vahakoski, M.Sc., Niina Santio, M.Sc. Undergraduate students: Sini Eerola, Heidi Ekman, Katja Männistö, Sofia Pruikkonen, Juho Virtanen. Description of the Project: The studies of our research group focus on the signaling pathways regulated by the oncogenic Pim family of serine/threonine-specific protein kinases. We have shown that the three highly homologous members of this family are expressed in partially overlapping patterns, mainly in cells of the immune or the nervous system. In hematopoietic cells, pim expression can be induced by multiple cytokines and also by some hormones, suggesting a role for Pim kinases in signal transduction initiated by cytokine or hormone receptors. When overexpressed in lymphoid tissues of transgenic mice, pim genes promote lymphomagenesis, especially in cooperation with other oncogenes that either enhance cell proliferation (myc) or cell survival (bcl-2). We and others have observed that in human cancer patients, elevated levels of pim-1 mRNA and protein can be found in leukemias, lymphomas and solid tumors such as prostate cancer. Most recently we have noticed that pim-1 overexpression promotes radioresistance in patients suffering from squamocellular head and neck carcinomas. We have previously shown that Pim-1 stimulates activities of several cellular or viral transcription factors such as Myb, NFATc, EBNA2 as well as RUNX family members. Most recently also LANA, the latency-associated nuclear antigen of Kaposi sarcoma-associated herpesvirus has been identified as a direct Pim substrate. We have also analysed expression of pim family genes during cytokinedependent T helper cell differentiation. Furthermore, we have shown that Pim kinases promote cytokine-independent survival and inhibit apoptosis by several mechanisms, including upregulated expression of the anti-apoptotic Bcl-2 protein and phosphorylation-induced inactivation of the pro-apoptotic Bad protein. Altogether, our studies based on domestic or international collaborations have had a major impact to the understanding of Pim kinase activities in both normal and transformed cells and have explained why Pim kinases can 58 From left to right: Eeva Rainio, Päivi Koskinen, Heidi Ekman, Jouko Sandholm, Niina Santio, Sofia Pruikkonen and Riitta Vahakoski. 59 specific small molecule inhibitors, which appear to be great tools for our research, but may also have therapeutic value. Using these inhibitors, we have recently revealed a novel role for Pim kinases in stimulation of cancer cell migration and invasion. Funding: Academy of Finland, Turku University Foundation Drug Discovery Graduate School. Selected Publications: Peltola, K.J., Hollmén, M., Maula, S.M., Rainio, E.M., Ristamäki, R., Luukkaa, M., Sandholm, J., Sundvall, M., Elenius, K., Koskinen, P.J., Grenman, R. and Jalkanen, S. (2009) Pim-1 kinase expression predicts radiation response in squamocellular carcinoma of head and neck and is under the control of epidermal growth factor receptor. Neoplasia 11: 629-636. Cheng, F., Weidner-Glunde, M., Varjosalo, M., Rainio, E.M., Lehtonen, A., Schulz, T.F., Koskinen, P.J., Taipale, J. and Ojala, P.M. (2009) KSHV reactivation from latency requires Pim-1 and Pim-3 kinases to inactivate the latency-associated nuclear antigen LANA. PLoS Pathogens, 5, e1000324 Aho, T.L.T., Peltola, K.J. and Koskinen, P.J. (2006) Pim-1 kinase phosphorylates RUNX family transcription factors and enhances their activity. BMC Cell Biol. 7: 1-9. Aho, T.L.T., Lund, R., Ylikoski, E., Matikainen, S., Lahesmaa, R. and Koskinen, P.J. (2005) Expression of human pim family genes is selectively upregulated by cytokines promoting Th1, but not Th2 cell differentiation. Immunol. 116: 82-88. Glazova, M., Aho, T.L.T., Palmetshofer, A., Murashov, A., Scheinin, M. and Koskinen, P.J. (2005). Pim-1 kinase enhances NFATc activity and neuroendocrine functions in PC12 cells. Mol. Brain Res. 138: 116-123. Rainio, E.M., Ahlfors, H., Carter, K., Ruuska, M., Matikainen, S., Kieff, E. and Koskinen, P.J. (2005) Pim kinases are upregulated by Epstein-Barr virus infection and enhance EBNA2 activity. Virol. 333: 201-206. Peltola, K.J., Paukku, K., Aho, T.L.T., Ruuska, M., Silvennoinen, O. and Koskinen, P.J. (2004). Pim-1 kinase inhibits Stat5-dependent transcription via its interactions with SOCS1 and SOCS3. Blood 103: 3744-3750. Aho, T.L.T., Sandholm, J., Peltola, K.J., Mankonen, H.P., Lilly, M. and Koskinen, P.J. (2004) Pim-1 kinase promotes inactivation of the pro-apoptotic Bad protein by phosphorylating it on the Ser112 gatekeeper site. FEBS Lett. 571: 43-49. Yan, B., Zemskova, M., Kraft, A., Koskinen, P.J. and Lilly, M. (2003). The Pim-2 kinase phosphorylates Bad on serine-112 and reverses Bad-induced cell death. J. Biol. Chem. 278: 45358-45367. Rainio, E.M., Sandholm, J. and Koskinen, P.J. (2002). Transcriptional activity of NFATc1 is enhanced by the Pim-1 kinase. J. Immunol.168: 1524-1527. Eichmann, A., Yuan, L., Bréant, C., Alitalo, K. and Koskinen, P.J. (2000). Developmental expression of Pim kinases suggests functions also outside of the hematopoietic system. Oncogene 19: 1215-1224. 60 MOLECULAR AND SYSTEMS IMMUNOLOGY AND STEM CELL BIOLOGY Principal investigator: Riitta Lahesmaa, M.D., Ph.D., Professor, Turku Centre for Biotechnology, BioCity, Tykistökatu 6A, FI-20521 Turku, Finland. Tel. +358-2-333 8601, Fax. +358-2-333 8000. Email: [email protected] Biography: Riitta Lahesmaa received her M.D. in 1984 and Ph.D. in 1987 from the University of Turku, and was appointed Docent in Immunology in 1990. She was a postdoctoral fellow at Stanford University Medical Center with Professor Lawrence Steinman during the years 1990-1993 (NIH Fogarty Fellowship). In 1994 she moved to Syntex Research Institute (later Roche Bioscience) in Palo Alto, California. As a Principal Scientist she focused on lymphocyte signaling and drug discovery with state-of-the-art functional genomics tools. In 1998 she was appointed Director of Turku Centre for Biotechnology. In 2009 she carried out research in Professor Anjana Rao’s laboratory in Immune Disease Institute, Harvard Medical School, Boston. She also directs BioCity Turku Systems Biology Research Program since 2000. Personnel: Senior scientists/ Post-doctoral researchers: Sanna Edelman, Ph.D., Laura Elo-Uhlgren, Ph.D.; Bhawna Gupta, Ph.D.; Riikka Lund, Ph.D.; Robert Moulder, Ph.D.; Juha-Pekka Pursiheimo, Ph.D.; Sunil Raghav, Ph.D.; Omid Rasool, Ph.D.; Emaheswa Reddy, Ph.D.; Johanna Tahvanainen, Ph.D. Visiting Scientists: David Hawkins, Ph.D. (Ludwig Institute of Cancer Research, San Diego, USA), Kanury Rao, Ph.D., (Director, Immunology Group at ICGEB, New Delhi, India); Brigitta Stockinger, Ph.D. (Principal Investigator, Division of Molecular Immunology, NIMR, London, UK) Graduate students: Helena Ahlfors, M.Sc.; Sanna Filen, M.Sc. Henna Järvenpää, M.Sc.; Juha Korhonen, M.D.; Minna Kyläniemi, M.Sc. Tapio Lönnberg, M.Sc.; Elisa Närvä, M.Sc.;Mirkka Heinonen, M.Sc. Nelly Rahkonen, M.Sc.; Soile Tuomela, M.Sc. Subhash Tripathi, M.Tech, M.Sc. Technicians: Marjo Hakkarainen, Sarita Heinonen, Päivi Junni Undergraduate students: Suvi Kantola, Kaarina Ranta, Marjo Linja, Joel Nyström, Juuso Nästi, Verna Salonen Description of the project : Our research is focused on molecular systems immunology and stem cell biology. We use holistic genome and proteome wide methods and systems biology to reveal molecular mechanisms of cell signaling, transcriptional and epigenetic programs that determine cell differentiation and fate. These approaches are exploited to understand molecular mechanisms of human immune mediated diseases and certain types of cancer to provide novel therapeutic means to modulate harmful cellular and immune responses. 61 T helper cell activation and differentiation to functionally distinct subsets Selective activation of T helper (Th) cell subsets plays an important role in the pathogenesis of human allergy and inflammatory diseases. Dissecting pathways and regulatory networks leading to the development of Th1, Th2, Th17 or regulatory T cells (Treg) is essential to understand the pathogenesis of allergy and inflammatory diseases. Th2 cytokines lead to a series of inflammatory processes characteristic for asthma and other atopic diseases whereas Th1 and Th17 cells play a role in the pathogenesis of autoimmune diseases (e.g. type I diabetes). Treg cells have an important role in inhibiting all these T effector cell functions. We have applied a holistic approach to identify genes involved in human Th cell differentiation. Detailed analysis of upstream T Cell Receptor (TCR)/key cytokine receptor induced differentiation will increase our understanding of these processes central for human health and disease and provide novel insights into new therapeutic interventions. STAT6 is known to be an essential upstream mediator of IL-4R signaling and Th2 differentiation. Importantly, we identified for the first time STAT6 target genes on a genome wide scale in human CD4+ T cells - only small fraction of which were previously known to be STAT-6 regulated. This study, published in Immunity, revealed that in human surprisingly high proportion, up to 80% of IL-4 induced response is STAT6 regulated revealing several new candidates for therapeutic intervention (Elo L et al. 2010). Our studies on IL-4 R signaling in lymphocytes also resulted in identification of new IL4R/STAT-6 regulated proteins in human and mice as well as mechanistic studies on their molecular functions (Aflakian N, et al. 2009, Moulder R. et al. 2010, Tuomela S. et al. 2009, Cho CH et al. 2009). Our results have led to novel hypotheses on the key factors involved in human Th cell differentiation (Lund et al., 2007, Rautajoki et al. 2007). Elucidating their functions further we discovered that ATF3 and SATB1 are important regulators of human Th cell differentiation. ATF3 promotes Th1 differentiation (Filen S. El al. 2010) whereas SATB1 regulates multiple genes during early Th cell differentiation (Ahlfors et al. 2010). Human embryonic stem cells (hESC) have a unique capacity to differentiate to any type of cell or tissue providing an enormous potential for therapeutic applications. Our recent results based on the use of high resolution microarray technology demonstrate that it is essential to monitor stem cell lines carefully to minimize the risk of malignancies in stem cell therapies. Our study published in Nature Biotechnology and highlighted in Nature Methods revealed that in prolonged culture human embryonic stem cells acquire chromosomal abnormalities and changes in gene expression, many of which are linked to cancer. (Närvä et al. 2010). Our goal is to elucidate the molecular mechanisms regulating self renewal and pluripotency of hESC and induced pluripotent stem cells (iPS). We have identified novel genes and signaling pathways characteristic for the pluripotent hESC and iPS cells based on a genome wide transcriptome analyses of hESC. Current work aims at further characterization and functional analysis of a panel of selected factors in the maintenance of undifferentiated status of hESC. Type 1 diabetes (T1D) is the most common metabolic-endocrine disorder in children in western countries and the annual incidence of T1D in Finland is record high. In almost all children, progression to clinical T1D is associated with the presence of β cell specific 62 autoantibodies. Clinical T1D occurs when 80-90% of the β cells have been destroyed. At this point T1D patient is dependent on a daily insulin substitution for the rest of his/her life and there is a high risk of developing acute and long-term complications. Development of early diagnostics would enable early therapy and possibly preventive treatments resulting in a significant reduction in the health care costs. Our objective is to study molecular mechanisms of T1D and to discover molecular markers that indicate development of autoimmunity and progression towards clinical T1D. Exploiting the unique biobank of the Type 1 Diabetes Prediction and Prevention Project in Finland (DIPP) we investigated transcriptomic profiles of prospective whole-blood samples from children who have developed T1D-associated autoantibodies and eventually clinical T1D. Gene-level investigation of the data showed systematic differential expression of 520 probesets. A network-based analysis revealed then a highly significant down-regulated network of genes involved in antigen presentation as well as T-cell receptor and insulin signaling. (Elo et al. 2010). Further studies include analysis of larger cohort of longitudinal samples using transciptomics, proteomics and integrating the data with our previous metbolomics results (Oresic et al. 2008). Funding: The Academy of Finland, The National Technology Agency of Finland (TEKES), EU 6th framework “ESTOOLS”, JDRF, The Sigrid Jusélius Foundation, The Finnish Cancer Organizations, Turku University Hospital Fund, Graduate Schools (TuBS, DDGS, ISB), University of Turku, Åbo Akademi University, European Research Council, EU 7th framework “SYBILLA”, EU 7th framework “DIABIMMUNE” EU 7th framework “NANOMMUNE”, EraSysBioPlus, European Research Council Collaborators: Harri Lähdesmäki (Tampere University of Technology and Aalto University, Reija Autio & Olli Yli-Harja (Tampere University of Technology ), Tero Aittokallio & Olli Nevalainen (UTU), Peter Andrews (University of Sheffield, UK) and the rest of EU FP6 ESTOOLS consortium (Altogether 20 partners), Outi Hovatta (Karolinska Institute, Stockholm, Sweden), Matej Oresic (VTT Technical Research Centre of Finland, Turku), Brigitta Stockinger (NIMR, London, UK and visiting professor at CBT), Kanury V.S. Rao (ICGEB, New Delhi, India and visiting professor at CBT), Anjana Rao (Immune Disease Institute, Harvard Medical School, Boston, MA, USA), Thomas Tushl (Rockefeller University, New York, NY, USA), Christopher Burge (MIT, Cambridge, MA, USA), David Goodlett (University of Washington, Seattle, WA, USA) , Matthias Gstaiger & Ruedi Aebersol (ETZ, Zürich, Switzerland) and the rest of EU FP7 SYBILLA partners (Altogether 14), Panu Jaakkola (Turku Centre for Biotechnology), Olli Simell, Jorma Ilonen & Heikki Hyöty, Juha Kere (Karolinska Institute, Stockholm, Sweden), Bing Ren (Ludwig Institute for Cancer Research, University of California, San Diego, USA), Mikael Knip (University of Helsinki) and the rest of EU FP7 DIABIMMUNE partners (Altogether 12), Bengt Fadeel (Karolinska Institute, Stockholm, Sweden ) and the rest of EU FP7 DIABIMMUNE partners (Altogether 14 partners). Selected Publications: Aflakian N, Ravichandran S, Sarwar Jamaal Md. S, Jarvenpää H, Lahesmaa, R, Rao KVS. (2009) Integration of signals from the 63 B-cell antigen receptor and the IL-4 receptor leads to a cooperative shift in the cellular response axis. Mol Biosyst. 5:1661-71. Ahlfors H, Limaye A, Elo-Uhlgrén L, Notani D, Gottimukkala K, Burute M, Tuomela S, Rasool O, Galande S* & Lahesmaa R*. (2010) SATB1 dictates expression of multiple genes including IL-5 involved in human T helper cell differentiation. *Equal contribution. Blood. 116:1443-53. Chen, Z., Lund, R., Aittokallio, T., Nevalainen, O. and Lahesmaa, R. (2003) Identification of early, positively and negatively regulated targets of STAT6 in IL-4 stimulated CD4+ T lymphocytes induced to polarize to Th2 cells. J Immunol. 171: 3627-3635. Cho SH, Goenka S, Henttinen T, Gudapati P, Reinikainen A, Lahesmaa R, Boothby M. (2009) PARP-14, a member of the B aggressive lymphoma (BAL) family, transduces survival signals in primary B cells. Blood. 113:2416-25. (2007) Systematic construction of gene coexpression networks with applications to human T helper cell differentiation process. Bioinformatics. 23: 2096-2103. Elo LL#, Järvenpää H#, Tuomela S#, Raghav S#, Ahlfors H, Laurila K, Gupta B, Lund RJ, Tahvanainen J, Hawkins RD, Orešic M, Lähdesmäki H, Rasool O, Rao KVS*, Aittokallio T*, Lahesmaa R. (2010) IL-4- and STAT6-mediated transcriptional regulation to initiate Th2 program in human T cells. Immunity 32:852-62#, * Equal contribution. Elo LL*, Mykkänen J*, Nikula T, Järvenpää H, Aittokallio T, Hyöty H, Ilonen J, Veijola R, Knip M, Simell O, Lahesmaa R. (2010) Genomewide gene expression profiling reveals early suppression of immune response pathways in prediabetic children. *Equal contribution. J Autoimmun. 35:70-6. Filén JJ, Filén S, Moulder R, Tuomela S, Ahlfors H, West A, Kouvonen P, Kantola S, Björkman M, Katajamaa M, Rasool O, Nyman TA, Lahesmaa R. (2009) Mol Cell Proteomics. 8:32-44. Filén S, Ylikoski E, Tripathi S, West A, Björkman M, Nyström J, Ahlfors H, Rao KVS, Coffey E, Rasool O, and Lahesmaa R. (2010) ATF3 is a Positive Regulator of Human IFNG Gene Expression. J Immunol. 184:4990-9. From left to right, first row: Juha-Pekka Pursiheimo, Emaheswa Reddy, Riikka Lund, Sarita Heinonen, Soile Tuomela, Sanna Edelman, second row: Omid Rasool, Emilia Engström, Nelly Rahkonen, Johanna Tahvanainen, Riina Plosila, Marjo Hakkarainen, Tapio Lönnberg, third row: Subhash Tripathi, Verna Salo, Riitta Lahesmaa. Hämäläinen, H., Zhou, H., Chou, W., Hashizume, H., Heller, R. and Lahesmaa R. (2001) Profiling Human Th1 and Th2 Differentiation by High-density Oligonucleotide Arrays. Genome Biology 2 (7): research0022. Kumar D, Srikanth R, Ahlfors H, Lahesmaa R, Rao K, (2007) Capturing cell-fate decisions from the molecular signatures of a receptor-dependent signaling response. Molecular Systems Biology. 3:150. Lund, R., Aittokallio, T., Nevalainen, O. and Lahesmaa, R. (2003) Identification of novel genes regulated by IL-12, IL-4 and TGFb during the early polarization of CD4+ lymphocytes. J. Immunol. 171: 5328-5336. 64 65 Lund, R., Chen, Z., Scheinin, J. and Lahesmaa, R. (2004) Early target genes of IL-12/STAT4 signaling in T helper cells. J. Immunol. 172: 6775-6782. Lund R*, Pykäläinen M*, Naumanen T, Dixon C, Chen Z, Ahlfors H, Tuomela S, Tahvanainen J, Scheinin J, Henttinen T, Rasool O, Lahesmaa R. (2007) Genome wide identification of Novel Genes Involved in Early Th1 and Th2 Cell Differentiation J. Immunol. 178:3648-60. Moulder R*, Lönnberg T*, Filén J-J, Elo L, Rainio E, Corthals G, Oresic M, Nyman TA, Aittokallio T, Lahesmaa R (2010) (*equal contribution). Quantitative Proteomics Analysis of the Nuclear Fraction of Human CD4+ Cells in the Early Phases of IL-4 Induced Th2 Differentiation. Mol Cell Proteomics. 9:1937-53. Närvä E, Autio R, Rahkonen N, Kong L, Harrison N, Kitsberg D, Borghese L, Itskovitz-Eldor J, Rasool O, Dvorak P, Hovatta O, Otonkoski T, Tuuri T, Cui W, Brüstle O, Baker D, Maltby E, Moore HD, Benvenisty N, Andrews PW, Yli-Harja O & Lahesmaa R. (2010) High resolution genome wide DNA analysis on a large panel of Human Embryonic Stem Cell lines reveals novel genomic changes associated with culture and affecting gene expression. Nat Biotechnol. 28:371-7. Oresic M, Simell S*, Sysi-Aho M*, Näntö-Salonen K*, SeppänenLaakso T*, Parikka V*, Katajamaa M*, Hekkala A, Mattila I, Keskinen P, Yetukuri L, Reinikainen A, Lähde J, Suortti T, Hakalax J, Simell T, Equal contribution. J Exp Med. 205:2975-84. * Rautajoki, K., Marttila, E., Nyman, T., Lahesmaa, R. (2007) Interleukin-4 inhibits caspase-3 by regulating several proteins in the Fas pathway during initial stages of human T helper 2 cell differentiation. Mol. Cell Proteomics. 6: 238-251. Tahvanainen J, Kallonen T, Lähteenmäki H, Heiskanen KM, Westermarck J, Rao KV, Lahesmaa R. (2008) Blood. 113:1268-77. Tuomela S, Rautajoki KJ, Moulder R, Nyman TA, Lahesmaa R. (2009) Identification of novel Stat6 regulated proteins in IL-4-treated mouse lymphocytes. Proteomics. 9:1087-98. PROTEIN CRYSTALLOGRAPHY Principal investigator: Anastassios C. Papageorgiou, Ph.D., Adjunct Professor in Biochemistry and Structural Biology Turku Centre for Biotechnology, BioCity, Tykistökatu 6A, FI-20521 Turku, Finland. Tel. +358-2-3338012, Fax +358-2-3338000. Email: [email protected] Biography: Tassos Papageorgiou (b. 1962) obtained his Ph.D. from the University of Athens in 1992. He was a postdoctoral fellow at the University of Oxford and University of Bath (UK). In May 2000, he joined the Centre for Biotechnology as senior scientist in protein crystallography Personnel: Graduate students: Prathusha Dhavala, Teemu Haikarainen, Sachin Wakadkar Undergraduate students: Simon Le Boulh, Pia Kinaret. Omid Mohammadi, Bishwa Subedi, Kristian Wecström Description of the projects: We use X-crystallography, molecular biology and biophysical techniques to study the structure and function of biological molecules. One of our major projects has been the Dps family of proteins that are widely spread among procaryotes and responsible for protection against oxidative stress due to their ability to oxidize and store iron. Although Dps proteins are structurally similar to ferritins, they form a spherical shell of 12 subunits instead of 24 and have a different ferroxidase center compared to that of ferritins. The crystal structures of Dps-like peroxide resistance protein (Dpr) from the pathogenic bacterium Streptococcus suis in the iron-free, the iron-bound and zinc-bound form have been recently determined. In addition, EXAFS experiments performed at EMBL Hamburg have provided detailed information of the geometry of the iron core and showed a ferrihydrite-like structure. Based on our recent results, a number of mutants have been generated to study the iron core formation using X-ray crystallography, microcalorimetry, EXAFS, magnetization and Mössbauer spectroscopy techniques. The structure of Dpr from Streptococcus pyogenes was determined in 2009 and structural studies on Helicobacter pylori neutrophilactivating protein (known as HP-NAP) were initiated to identify potential ligand binding sites. Studies on oxidative stress protection and detoxification mechanisms have been extended by determining high-resolution crystal structures of a tau family glutathione transferase (GST) from Glycine max in free form and in the presence of a substrate analogue. Importantly, the crystal structures revealed a novel site on the surface of the protein that may be utilised for storage and/ or transport of dangerous compounds for detoxification. Docking calculations were carried out to study the binding of diphenylether herbicides in the active site. Work is currently underway on chimeric GSTs or mutants created through directed evolution approaches to produce new GSTs with altered specificity for new applications in agriculture and biomedicine. Crystals of human GST-A1 have been grown in our lab for use in structure-assisted drug design efforts. In addition, the structure of a novel glutathione transferase 66 67 was determined by the SAD method using the anomalous signal of Br. The overall fold and the geometry of the active site suggest a new member of the glutathione transferase superfamily. Knock-out and microarray experiments are currently in progress to provide functional insights. Melissis, S.C., Papageorgiou, A.C., Labrou, N.E & Clonis, Y.D. (2010). Purification of moloney murine leukemia virus reverse transcriptase lacking RNase activity (M-MLVH-RT) on a 9-aminoethyladenine[1,6-diamine-hexane]-triazine selected from a combinatorial library of dNTP-mimetic ligands. J. Chromatogr. Sci. 48, 496-502. In the theme of enzyme function and stability, we continued our work on PhaZ7, an extracellular depolymerase involved in the degradation of poly(R)-hydroxyalkanoates, a group of thermoplastic polyesters considered as biodegradable substitutes for nondegradable plastics. The crystal structure of PhaZ7 depolymerase at atomic (1.2) Å resolution in the presence of the serine protease inhibitor PMSF was determined. A molecule of SO2 was found covalently bound at the active site of the enzyme, suggesting a preformed catalytic triad. Several mutants have been generated by our collaborators and characterized. Crystal structure determination is currently underway. The structure of Erwinia carotovora asparaginase at 1.4 Å, the second highest resolution for an L-asparaginase was determined. Asparaginases are widelyused enzymes in leukemia treatment for the last 30 years. However, asparaginases from Erwinia chrysanthemi and E. coli cause severe side-effects. Protein engineering efforts have been initiated for the Erwinia carotovora enzyme based on its 3D-structure to produce chimeric forms with reduced glutaminase activity (the main reason of the side-effects) and high specificity for L-asparagine. Work on the Atu (acyclic terpene utilization) catabolic pathway found in P. Aeruginosa has been initiated using a combination of X-ray crystallography, biophysics, molecular biology, homology modelling, computational and bioinformatics tools. Atu enzymes are involved in the metabolisn of acyclic terpenes that possess a great potential in biotechnology, for example in the food, drink and pharmaceutical industry. Haikarainen, T., Tsou, C.C., Wu, J.J. & Papageorgiou, A.C. (2010). Crystal structures of Streptococcus pyogenes Dpr reveal a dodecameric iron-binding protein with a ferroxidase site. J. Biol. Inorg. Chem. 15, 183-194. Selected publications: Wakadkar, S., Zhang,L.Q., Li, D.-C., Haikarainen, T., Dhavala, P. & Papageorgiou, A.C. (2010). Expression, purification and crystallization of Chetomium thermophilum Cu, Zn superoxide dismutase. Acta Cryst F (in press). Haikarainen, T., Tsou, C.C., Wu, J.J. & Papageorgiou, A.C. (2010). Structural characterization and biological implications of di-zink binding in the ferroxidase center of Strepococcus pyogenes Dpr. Bichem. Biophys. Res. Comm. 398, 361-365. Haikarainen, T. & Papageorgiou, A.C. (2010). Dps-like proteins: Structural and functional insights into a versatile protein family. Cell. Mol. Life Sci. 67, 341-351. Axarli, I., Georgiadou, C., Dhavala, P., Papageorgiou, A.C. & Labrou, N. (2010). Investigation of the role of conserved residues Ser13, Asn48 and Pro49 in the catalytic mechanism of the tau class glutathione transferase from Glycine max. Bioch. Biophys. Acta 1804, 662-667. Labrou, N., Papageorgiou, A.C. & Avramis, V.I. (2010). Structurefunction relationships and clinical applications of L-asparaginases. Curr. Med. Chem. 17, 2183-2195. Wakadkar, S., Hermawan, S., Jendrossek, D. & Papageorgiou, A.C. (2010). The crystal structure of PhaZ7 at atomic (1.2 Å) resolution reveals details of the active site and suggests a substrate-binding mode. Acta Cryst. F 66, 648-654. 68 Axarli, I. Dhavala, P., Papageorgiou, A.C. & Labrou, N.E. (2009). Crystallographic and functional characterization of the fluorodifeninducible glutathione transferase from Glycine max reveals an active site topography suited for diphenylether herbicides and a novel L-site. J. Mol. Biol. 385, 984-1002. Axarli, I. Dhavala, P., Papageorgiou, A.C. & Labrou, N.E. (2009). Crystal structrure of Glycine max glutathione transferase in complex with glutathione: investigation of the induced-fit mechanism operating by the tau class glutathione transferases. Biochem. J. 422, 247-256. Mitsiki, E., Papageorgiou, A. C., Iyer, S., Thiyagarajan, N., Prior, S. H., Sleep, D., Finnis, C. & Acharya, K. R. (2009). Structures of native human thymidine phosphorylase and in complex with 5-iodouracil. Biochem. Biophys. Res. Commun. 386, 666-670. Dhavala, P. & Papageorgiou, A.C. (2009). The crystal structure of Helicobacter pylori L-asparaginase at 1.4 Å resolution. Acta Crystallogr. D 65, 1253-1261. Havukainen, H., Haataja, S., Kauko, A., Pulliainen, A.T., Salminen, A., Haikarainen, T., Finne, J. & Papageorgiou, A.C. (2008). Structural basis of zinc- and terbium-mediated inhibition of ferroxidase activity in Dps ferritin-like proteins. Protein Sci. 17, 1513-1521 Papageorgiou, A.C., Posypanova, G.A., Andersson, C.A., Sokolov, N.N & Krasotkina, J. (2008) Structural and functional insights into Erwinia carotovora L-asparaginase. FEBS J. 275, 4306-4316 Dhavala, P., Krasotkina, J., Dubreuil, C. & Papageorgiou, A.C. (2008). Expression, purification and crystallization of Helicobacter pylori L-asparaginase. Acta Crystallogr Sect F Struct Biol Cryst Commun. 64, 740-742 Papageorgiou, A.C., Hermawan, S., Singh C.B. & Jendrossek, D. (2008) Structural basis of poly(3-hydroxybutyrate) hydrolysis by PhaZ7 depolymerase from Paucimonas lemoignei. J. Mol. Biol. 382, 1184-1194 Saarinen S., Kato, H., Uchiyama, T., Miyoshi-Akiyama, T. & Papageorgiou, A.C. (2007). Crystal structure of Streptococcus dysgalactiae-derived mitogen reveals a zinc-binding site and alterations in TcR binding. J. Mol. Biol. 373, 1089-1097 Weckström, K. & Papageorgiou, A.C. (2007). Lower consolute boundaries of the nonionic surfactant C(8)E(5) in aqueous alkali halide solutions: An approach to reproduce the effects of alkali halides on the cloud-point temperature. J Colloid Interface Sci. 310, 151-162 69 Zhao, J., Hayashi, T., Saarinen, S., Papageorgiou, A.C., Kato, H., Imanishi, K., Kirikae, T., Abe, R., Uchiyama, T. & MiyoshiAkiyama, T. (2007). Cloning, expression and characterization of the superantigen streptococcal pyrogenic exotoxin-G from Streptococcus dysgalactiae. Inf. Immun. 75, 1721-1729 Kauko, A., Pulliainen, A.T., Haataja, S., Meyer-Klaucke, W., Finne, J. & Papageorgiou, A.C. (2006). Iron incorporation in Streptococcus suis Dps-like peroxide resistance protein Dpr requires mobility in the ferroxidase center and leads to the formation of a ferrihydritelike core. J. Mol. Biol. 364: 97-109 Papageorgiou, A.C., Saarinen, S., Ramirez-Bartutis, R., Kato, H., Uchiyama, T., Kirikae, T. & Miyoshi-Akiyama, T. (2006). Expression, purification and crystallisation of Streptococcus dysgalactiaederived mitogen. Acta Crystallogr. F. 62: 242-244 Kapetaniou, E.G., Thanassoulas, A., Dubnovitsky, A.P., Nounesis, G. & Papageorgiou, A.C. (2006). The effect of pH on the structure and stability of Bacillus circulans ssp. alkalophilus phosphoserine aminotransferase: Thermodynamic and crystallographic studies. Proteins: Struct. Funct. Bioinform. 63: 742-753 Pulliainen, A.T., Kauko, A., Haataja, S., Papageorgiou, A.C. & Finne, J. (2005). Dpr/Dps miniferritin: Insights into the mechanism of iron incorporation and evidence for a central role in cellular iron homeostasis in Streptococcus suis. Mol. Microbiol. 57, 10861100. Kapetaniou, E.G, Braaz, R., Jendrossek, D. & Papageorgiou, A.C. (2005). Crystallization and preliminary X-ray analysis of a novel thermoalkalophilic depolymerase (PhaZ7) from Paucimonas lemoignei. Acta Crystallogr. F 61: 479-481 Wikman, L.E.K., Krasotkina, J., Kuchumova, A., Sokolov, N.N. & Papageorgiou, A.C. (2005). Crystallization and preliminary crystallographic analysis of L-asparaginase from Erwinia carotovora. Acta Crystallogr. F 61: 407-409 Dubnovitsky,. A.P., Ravelli, R.B.G., Popov, A.N. & Papageorgiou, A.C. (2005). Strain relief at the active site of phosphoserine aminotransferase induced by radiation damage. Protein Sci. 14: 1498-1507 From left to right, sitting: Bishwa Subedi, Teemu Haikarainen, back row: Tassos Papageorgiou, Sachin Wakadkar, Prathusha Dhavala. Mialon A., Sankinen, M., Söderström, H., Junttila, T.T., Holmström, T., Koivusalo, R., Papageorgiou, A.C., Johnson, R.S., Hietanen, S., Elenius, K. & Westermarck, J. (2005). DNA topoisomerase I is a cofactor for c-Jun in the regulation of EGFR expression and cancer cell proliferation. Mol. Cell. Biol. 25: 5040-5051 Dubnovitsky, A.P., Kapetaniou, E.G. & Papageorgiou, A.C. (2005). Enzyme adaptation to alkaline pH: Atomic resolution (1.08 Å) structure of phosphoserine aminotransferase from Bacillus alkalophilus. Protein Sci. 14: 97-110 70 71 Bioinformatics unit Group leader (Structural Bioinformatics): Konstantin Denessiouk, Ph.D., Docent in Biochemistry. Bioinformatics Group leader. Centre for Biotechnology, Tykistökatu 6, BioCity 5th floor, Turku, 20520 Turku. E-mail: [email protected] Personnel: Bhanupratap Singh Chouhan. Group leader (High-throughput Bioinformatics): Attila Gyenesei, Ph.D., Senior Scientist, Group leader, Turku Centre for Biotechnology, BioCity, Tykistökatu 6A, P.O. Box 123, FIN-2050 Turku, Finland. Tel. +3582-333 8634 Fax +358-2-333 8000. E-mail: [email protected] Personnel: Sini Junttila, Asta Laiho, Leena Kytömäki, Seppo Tamminen. Description of the Project (2008-2009): Tremendous amounts of data generated by high-throughput technologies in genomics and transcriptomics like DNA microarray and next-generation sequencing instruments that demand in-depth analysis present unprecedented new challenges to data analyst and biomedical researchers. The High-throughput bioinformatics (HTB) group is engaged in the ongoing development of advanced analysis tools and research on generating novel approaches for the analysis of high-throughput data sets. Moreover, in close collaboration with the Finnish Microarray and Sequencing Centre (FMSC) we provide services in the analysis of DNA microarray and next generation sequencing data produced within the Centre. Members of the HTB group participate in the initial project meetings, held at the beginning of each FMSC project, to assist the researchers with designing the experiment to guarantee the optimal experimental setup. Our bioinformatics services include data analysis of various data types such that gene and exon-specific expression data, ChIPchip and ChIP-seq data, SNP genotyping data, etc. Additionally, our group offers consultation and training for researchers to ensure that they benefit from the computational tools used in the analysis of biological data sets. Our aim is to educate and thereby enable researchers to work on their own data, which allows them to take the full benefit of their deep biological knowledge into consideration during the data analysis. Various courses are organized by our team each year to cover different aspects of data analysis. The Bioinformatics Unit provides support for Structural Bioinformatics and Chemical Informatics (in conjunction with the Structural Bioinformatics Laboratory, lead by Prof. Mark S. Johnson at the Åbo Akademi University); and separately, support for projects and development of high-throughput screening (HTS) of natural molecules (in conjunction with Prof. Pia Vuorela, Department of Biosciences, Åbo Akademi University). The Structural Bioinformatics Group has its main expertise in (a) computerbased analysis of protein-protein and protein-ligand interactions; (b) computer-aided prediction and intelligent molecular modeling and design; (c) computer-based ligand docking and analysis; (d) quantum chemistry, molecular dynamics; and (d) analysis of effects of molecular recognition and mutations on protein function. During year 2009, in collaboration with Prof. Jyrki Heino, University of Turku and Prof. Mark S. Johnson, Åbo Akademi University, the collaboration has been started in in-depth structural analysis 72 of human integrin beta-propeller domains and identification of structural features of integrins that distinguish human integrins from the other protein superfamilies having the same fold. Several matching sequences in bacteria that aligned surprisingly well with the integrin alpha subunits were identified (Johnson et al., 2009; Chouhan et al., 2010, ready to be submitted), giving a new insight into existence and evolution of the integrin-like proteins in bacteria. Additionally, our on-going research were focused on prediction of effects of splice variation on protein function (in collaboration with Prof. Riitta Lahesmaa, Turku Centre for Biotechnology), analysis of effects of molecular recognition and mutations on protein function in macromolecular receptor ErbB4 complexes (in collaboration with Dr. Klaus Elenius, University of Turku). Separately, the group guides individual training of MSc students, in collaboration with the Structural Bioinformatics Laboratory (Åbo Akademi University), and leads a Ph.D. student in Bioinformatics and Computational Biology within the National Graduate School of Informational and Structural Biology (Åbo Akademi University). Funding: The Åbo Akademi University; The National Graduate School in Informational and Structural Biology (ISB). Collaborators: Riitta Lahesmaa (Turku Centre for Biotechnology), Mark Johnson (Åbo Akademi University), Dr. Klaus Elenius (University of Turku); Prof. Jyrki Heino (University of Turku). Selected Publications: Chouhan B, Denesyuk A, Heino J, Johnson MS Denessiouk K. (2010) Conservation of the human integrin-type beta-propeller domain in bacteria. Ready to be submitted. Kankare M, Salminen T, Laiho A, Vesala L, Hoikkala A. Changes in gene expression linked with adult reproductive diapause in a northern malt fly species: a candidate gene microarray study. BMC Ecol. 2010 Feb 1;10:3.PMID: 20122138 Sirén A, Polvi A, Chahine L, Labuda M, Bourgoin S, Anttonen AK, Kousi M, Hirvonen K, Simola KO, Andermann E, Laiho A, Soini J, Koivikko M, Laaksonen R, Pandolfo M, Lehesjoki AE. Suggestive evidence for a new locus for epilepsy with heterogeneous phenotypes on chromosome 17q. Epilepsy Res. 2010 Jan;88(1):6575. Epub 2009 Nov 14.PMID: 19914042 Huvila J, Brandt A, Rojas CR, Pasanen S, Talve L, Hirsimäki P, Fey V, Kytömäki L, Saukko P, Carpén O, Soini JT, Grénman S, Auranen A. Gene Expression profiling og endometrial adenocarcinomas reveals increased apolipoprotein E expression in poorly differentiated tumors. Int J Gynecol Cancer. 2009 Oct;19(7):1226-31. Oksala N, Levula M, Airla N, Pelto-Huikko M, Ortiz RM, Järvinen O, Salenius JP, Ozsait B, Komurcu-Bayrak E, Erginel-Unaltuna N, Huovila AP, Kytömäki L, Soini JT, Kähönen M, Karhunen PJ, Laaksonen R, Lehtimäki T. ADAM-9, ADAM-15, and ADAM-17 are upregulated in macrophages in advanced human atherosclerotic plaques in aorta and carotid and femoral arteries--Tampere vascular study. Ann Med. 2009;41(4):279-90. Levula M, Airla N, Oksala N, Hernesniemi JA, Pelto-Huikko M, Salenius JP, Zeitlin R, Järvinen O, Huovila AP, Nikkari ST, Jaakkola 73 O, Ilveskoski E, Mikkelsson J, Perola M, Laaksonen R, Kytömäki L, Soini JT, Kahonen M, Parkkinen J, Karhunen PJ, Lehtimäki T. ADAM8 and its single nucleotide polymorphism 2662 T/G are associated with advanced atherosclerosis and fatal myocardial infarction: Tampere vascular study. Ann Med. 2009 Jul 2:1-11. Johnson MS, Lu N, Denessiouk K, Heino J, Gullberg D. (2009). Integrins during evolution: Evolutionary trees and model organisms. Biochim Biophys Acta. 1788: 779-789. Fan YM, Karhunen PJ, Levula M, Ilveskoski E, Mikkelsson J, Kajander OA, Järvinen O, Oksala N, Thusberg J, Vihinen M, Salenius JP, Kytömäki L, Soini JT, Laaksonen R, Lehtimäki T. Expression of sterol regulatory element-binding transcription factor (SREBF) 2 and SREBF cleavage-activating protein (SCAP) in human atheroma and the association of their allelic variants with sudden cardiac death. Thromb J. 2008 Dec 30;6:17. Cloke B, Huhtinen K, Fusi L, Kajihara T, Yliheikkilä M, Ho KK, Teklenburg G, Lavery S, Jones MC, Trew G, Kim JJ, Lam EW, Cartwright JE, Poutanen M, Brosens JJ. The androgen and progesterone receptors regulate distinct gene networks and cellular functions in decidualizing endometrium.. Endocrinology. 2008 Sep;149(9):4462-74. Akerfelt M, Henriksson E, Laiho A, Vihervaara A, Rautoma K, Kotaja N, Sistonen L. Promoter ChIP-chip analysis in mouse testis reveals Y chromosome occupancy by HSF2. Proc. Natl. Acad. Sci. U.S.A.. 2008 Aug;105(32):11224-9 pathways due to SKI-1/S1P inhibition in HepG2 cells.. DNA Cell Biol. 2007 Nov;26(11):765-72 Rodriguez A, Hilvo M, Kytömäki L, Fleming RE, Britton RS, Bacon BR, Parkkila S. Effects of iron loading on muscle: genome-wide mRNA expression profiling in the mouse. BMC Genomics. 2007 ;8():379-0 De Windt A, Rai M, Kytömäki L, Thelen KM, Lutjohann D, Bernier L, Davignon J, Soini J, Pandolfo M, Laaksonen R. Gene set enrichment analyses revealed several affected pathways in Niemann-pick disease type C fibroblasts.. DNA Cell Biol. 2007 Sep;26(9):665-71 Anckar J, Hietakangas V, Denessiouk K, Thiele DJ, Johnson MS, Sistonen L. (2006). Inhibition of DNA binding by differential sumoylation of heat shock factors. Mol Cell Biol. 26: 955-964. Poukkula M, Kaunisto A, Hietakangas V, Denessiouk K, Katajamäki T, Johnson MS, Sistonen L, Eriksson JE. (2005). Rapid turnover of c-FLIPshort is determined by its unique C-terminal tail. J Biol Chem. 280: 27345-27355. Denessiouk KA, Johnson MS, Denesyuk AI. (2005). Novel CaNN structural motif for protein recognition of phosphate ions. J Mol Biol. 345: 611-629. F.P. Pach, A. Gyenesei, and J. Abonyi. Visualization of fuzzy association rules. Journal of Visual Languages and Computing, Elsevier, in press, Available online, 2008. F.P. Pach, A. Gyenesei, and J. Abonyi. MOSSFARM: Model structure selection by fuzzy association rule mining. Journal of Intelligent and Fuzzy Systems 19(6): 399-407, 2008. F.P. Pach, A. Gyenesei, and J. Abonyi. Compact fuzzy association rule based classifier. Expert Systems with Applications 34(4): 24062416, 2008. Xhaard H, Backström V, Denessiouk K, Johnson MS. (2008). Coordination of Na(+) by monoamine ligands in dopamine, norepinephrine, and serotonin transporters. J Chem Inf Model. 48: 1423-1437. Denessiouk KA, Denesyuk AI, Johnson MS. (2008). Negative modulation of signal transduction via interleukin splice variation. Proteins. 71: 751-770. A. Gyenesei, U. Wagner, S. Barkow-Oesterreicher, E. Stolte, and R. Schlapbach. Mining co-regulated gene profiles for the detection of functional associations in gene expression data. Bioinformatics 23(15): 1927–1935, 2007. De Windt A, Rai M, Bernier L, Thelen K, Soini J, Lefebvre C, Chintawar S, Lavigne J, Saarinen L, Kytömäki L, Munzer JS, Lujohann D, Pandolfo M, Davignon J, Seidah NG, Laaksonen R. Gene set enrichment analysis reveals several globally affected 74 75 Cell fate Principal investigator: Cecilia Sahlgren, Ph.D., Docent in Cell and Development Biology, (Åbo Akademi University). Academy Research Fellow, Turku Centre for Biotechnology, Åbo Akademi and Turku University, BioCity, Tykistokatu 6B, FI-20521 Turku, Finland. Tel. +358-2-3338611, Fax. +358-2-3338000. Email: Biography: Cecilia Sahlgren received her Ph.D. from Turku Centre of Biotechnology, Åbo Akademi University December 2002. She was appointed research fellow at the Department of Biology at Åbo Akademi University from 2003-2005. 2005-2007 she was a postdoctoral fellow in Prof. Urban Lendahls lab at the Department of Cell and Molecular Biology at the Karolinska Institute. 2008 she was appointed senior research fellow (attending the position as professor of Biology) at Åbo Akademi University. In 2009 she founded the Cell fate group at the Turku Centre for Biotechnology. She currently holds an Academy of Finland Research Fellow position Personnel: Senior Scientist: Cecilia Sahlgren, Ph.D. Post-doctoral researcher: Veronika Mamaeva Graduate Students: Marika Hietamäki, M.Sc., Sebastian Landor, M.Sc, Anders Mutvei, M.Sc (KI), Laurel Tabe Bate-Eya, M.Sci Undergraduate Students: Daniel Antfolk, B.Sc, Christian Antila, B.Sc, Cecilia Granqvist, B.Sc, Rasmus Niemi Description of project: Cell-cell communication in development and disease: Targeting the Notch signaling pathway The main focus of our research is directed at elucidating the basic molecular principles of the signaling mechanisms that regulate cell fate choices during stem cell differentiation, and how disturbances in these mechanisms link to cancer. Another important goal is to develop technology to specifically monitor and tune these signals at will in specific cell populations, in order to steer stem cell fate and curtail oncogenic activities. The main focus is the role and regulation of the evolutionary conserved Notch signaling pathway, a key regulator of stem cell function and tumorigenesis. The main objectives of our research are to understand i) how the cellular microenvironment influences Notch signaling activities and how this impinges on cell identity and function, ii) how Notch signaling interlinks with other signaling mechanisms to fine tune and modulate the cellular response, iii) how intracellular temporal and spatial control of Notch signaling activities are achieved and to iv) develop technology platforms to regulate Notch signaling in targeted cell populations and for bioimaging of cellular functions in vivo. We have recently identified a Notch-hypoxia crosstalk of relevance for tumor progression (Sahlgren et al., 2008), and are currently participating in a project aimed at elucidating the Notch-hypoxia transcriptome to gain insight in how such a crosstalk is manifested on the transcriptome level and to obtain a molecular platform to better understand the intersection between the two signaling cascades in normal development and cancer (Main et al., 2010). We have shown that Notch signaling converts hypoxia inherent to the tumor microenvironment into epithelial mesenchymal transition (EMT) 76 required for the hypoxia-induced invasiveness of epithelial tumor cells. The link between Notch and hypoxia in tumor progression highlights the Notch pathway as an interesting therapeutic target in cancer. The identified Hypoxia-Notch-EMT axis also emphasizes the importance of understanding how Notch signaling becomes derailed and how this is manifested in various aspects of tumor progression. Half of the breast cancers have reduced levels of Numb, a Notch antagonist, and Notch expression is associated with poor prognosis. However, the specific role of Notch in breast cancer is yet unclear. We have created 3D cellular models and mouse models of breast cancer via orthotopic xenotransplantation (OX) using breast cancer cells expressing different and tunable levels of Notch activity to address these questions. Interaction between key signaling mechanisms is important to generate the diversity in signaling output required for proper control of cellular differentiation and function. Notch crosstalks with other major signaling pathways that modulate the signaling outcome. We have contributed to working out the relationship between Notch and the Notch antagonist, Numb (Chapman et al 2006) and to establishing data on a PDGF-Notch signaling crosstalk of relevance for vascular smooth muscle cell differentiation and function (Jin et al., 2008). Current research focus includes interactions between Notch receptors and ligands and the intermediate filament cytoskeleton, and how these interactions determine trafficking, signaling activity and Notch-driven stem cell functions. We aim to extend our studies on Notch signaling crosstalk and implement proteomics and mass-spec analysis to identify novel Notch binding proteins and posttranslational modifications of Notch. A systematic attempt to characterize how posttranslational modifications of Notch affect signaling and to identify interacting proteins is likely to reveal novel modes of regulation of Notch signaling. The Notch pathway is highlighted as an interesting therapeutic target, and developing strategies for local containment of Notch inhibitors to the primary tumor would be a productive way towards improving cancer therapy. Specific control over Notch activity is also of interest for regulating stem cell differentiation and regenerative therapy. We have recently described an approach to specifically target cell populations with nanoparticles and this technology is being further developed with the aim to specifically deliver Notch inhibitors to tumor cells to prevent cancer progression (Rosenholm et al 2009 a,b,c, Rosenholm et al., 2010). The developed technology provides a system for efficient and tight dose control of Notch signaling activity in a cell specific manner. In addition to the obvious therapeutic value, such cell-specific action and tight dose control of the Notch pathway should provide a useful tool for analyses of the biological output of the pleiotropic and dose-dependent Notch pathway. Further aims include developing the technology for a precise control of Notch driven stem cells functions and for imaging of stem cell functions and behaviour in vivo. Funding: The Academy of Finland, Åbo Akademi University (CoE in cell stress and ageing), Turku Graduate School of Biomedical Sciences, Magnus Ehrnrooth’s Stiftelse, Sigrid Jusélius Foundation, Finnish Cancer Organizations, the Tor Joe och Pentti Borgs Foundation (Åbo Akademi University). Collaborators: Prof. Milos Pekny (Sahlgrenska Academy at Göteborg University), 77 Prof. John Eriksson (Turku Centre for Biotechnology). Prof. Urban Lendahl (Karolinska Institute), Prof. Lucio Miele (Loyola Medical University, Chigaco), Ph.D Susumu Imanishi (Turku Centre for Biotechnology), Prof. Lea Sistonen (Turku Centre for Biotechnology). Dr.Tech Jessica Rosenholm (Laboratory for Physical Chemistry, Åbo Akademi, Turku), Prof. Mika Linden (Dept of Chemistry, Ulm University, Germany). Sahlgren C and Lendahl U. (2006) Notch, stem cell control and integration with other signaling mechanisms Regenerative Medicine 1 (2):195-20 Selected Publications (#equal contribution): Jessica M. Rosenholm, Emilia Peuhu, Laurel Tabe Bate-Eya, John E. Eriksson, Cecilia Sahlgren#, Mika Lindén#. Cancer-Cell Specific Induction of Apoptosis using Mesoporous Silica Nanoparticles as Drug Delivery Vectors Small 2010 Jun 6;6(11):1234-41.# equal coauthor contribution Aurelie de Thonel, Saima E. Ferraris, Hanna-Mari Pallari, Susumu Y. Imanishi, Vitaly Kochin, Tomohisa Hosokawa, Shin-ichi Hisanga, Cecilia Sahlgren, and John E. Eriksson. PKCζ regulates CDK5/p25 signaling during myogenesis in press (2010) MBC 21:1423-34 Heather Main*, Kian Leong Lee*, Henry Yang, Saija HaapaPaananen, Henrik Edgren, Shaobo Jin, Cecilia Sahlgren, Olli Kallioniemi, Lorenz Poellinger, Bing Lim and Urban Lendahl. Integration between Notch- and hypoxia-induced transcriptomes in embryonic stem cells. (2010) Exp Cell Res. 316:1610-24 Jessica M Rosenholm, Dr., Emilia Peuhu, M.Sc., John Eriksson, Prof. Dr., Cecilia Sahlgren, Dr. #, Mika Linden, Dr#. Targeted Intracellular Delivery of Hydrophobic Agents using Mesoporous Hybrid Silica Nanoparticles as Carrier Systems (2009) Nano Letters 9:3308-11 # equal co-author contribution Jessica Rosenholm, Cecilia Sahlgren, Mika Lindén. Cancer cellspecific targeting of and targeted delivery by mesoporous silica nanoparticles. (2010) Highlight to Journal of Material Chemistry 14:2707-2713 Jessica M Rosenholm, Dr., Annika Meinander, Dr., Emilia Peuhu, M.Sc., Rasmus Niemi, Mr., John Eriksson, Prof. Dr., Cecilia Sahlgren, Dr. #, Mika Linden, Dr#. Targeting of porous hybrid silica nanoparticles to cancer cells. (2009) ACSNano 3:197-206 # equal co-author contribution Shaobo Jin, Emil M. Hansson, Saara Ihalainen, Cecilia Sahlgren, Marc Baumann, Hannu Kalimo and Urban Lendahl. Notch signaling regulates PDGF-receptorβ expression in vascular smooth muscle cells. (2008) Circulation research 102:1483-91 Cecilia Granqvist, Rasmus Niemi, Veronika Mamaeva, Cecilia Sahlgren, Sebastian Landor, Christian Antila, Daniel Antfolk. Missing from the picture: Marika Hietamäki. Sahlgren, C, Gustafsson, M, Jin, S, Poellinger, L and Lendahl, U. Notch signaling mediates hypoxia induced tumor cell migration and invasion. (2008) Proceedings of National Academy of Sciences of the United States of America 105:6392-7. Gavin Chapman#, Lining Liu#, Cecilia Sahlgren, Camilla Dahlqvist, and Urban Lendahl. High levels of Notch signaling downregulate Numb and Numblike. (2006) Journal of Cell Biology, 175(4):53540. # authors contributed equally 78 79 Targeting strategies for gene therapy Principal investigator: Mikko Savontaus, M.D., Ph.D. Address: Turku Centre for Biotechnology, Biocity, Tykistökatu 6B, P.O. Box 123, FI-20521 Turku, Finland. Tel. +358 2 333 8025, Fax +358 2 333 8000. Email: [email protected] Biography: Mikko Savontaus (b. 1970) received his M.D. in 1996 and Ph.D. in 1997 from the University of Turku. He was a postdoctoral fellow at the Institute for Gene Therapy and Molecular Medicine at Mount Sinai School of Medicine in New York during 1999-2002. He is currently a senior scientist at the Turku Centre for Biotechnology as well as a specialist in internal medicine at the Department of Medicine at Turku University Hospital. Personnel: Graduate students: Raine Toivonen, M.Sc., Kim Eerola, M.Sc. Undergraduate student: Minttu Mattila Description of the project: Gene therapy is rapidly developing into a novel biomedical discipline that could have a major impact on health and healthcare in the 21st century. Traditionally gene therapy has been envisioned as a means to cure monogenic diseases with precisely defined genetic defects. However, recent clinical trials have demonstrated that gene therapy for complex multigenic disorders such as cardiovascular diseases and cancer are especially promising and may become a routine treatment modality in the near future. On the other hand, these trials have demonstrated that technical advances in gene therapy vector development are a key issue in developing clinically applicable gene therapy approaches. Our laboratory endeavors to tackle this problem of developing improved gene therapy vectors for cardiovascular diseases and cancer by attempting to meet two objectives: 1. The expression of therapeutic genes must be tightly regulated (transcriptional targeting). 2. The tropism of the gene delivery vector must be restricted to the target tissue (transductional targeting). Such targeted vectors will increase efficacy and diminish the possibility of side effects by limiting transgene expression to the target cell population. In our previous work we have constructed conditionally replicating adenoviruses (CRADs) targeting tumor endothelial cells and have demonstrated that these vectors are able to specifically replicate in dividing endothelial cells and destroy tumor vasculature. In addition, we have used a similar strategy to target tumor cells via the telomerase reverse transcriptase promoter. We have also demonstrated that a hybrid Ad5/35 adenovirus, where the fiber gene of adenovirus serotype 5 has been replaced with the fiber from serotype 35, is highly efficient in infecting endothelial cells. dilated cardiomyopathy. Ultrasound-guided injections are used to analyze the efficacy and toxicity of our targeted vectors after intramyocardial injection. Novel vectors with improved transcriptional and transductional efficiency for target cells will be constructed by combining hybrid serotype vectors with transcriptional targeting. In addition, we are utilizing lentivirus technology for long-term expression of therapeutic genes in the heart for heart failure and hypertension. Our ultimate goal is to develop gene therapy vectors for use in clinical trials by combining these approaches. Funding: Academy of Finland, Finnish Medical Foundation, Turku University Hospital Selected publications: Histochem Cell Biol. 133(3):349-57. Toivonen, R., Suominen, E., Grenman, R. and Savontaus, M. (2009) Retargeting Improves the Efficacy of a TelomeraseDependent Oncolytic Adenovirus for Head and Neck Cancer. Oncology Reports 21: 165-171 Suominen, E., Toivonen, R., Grenman, R. and Savontaus, M. (2006) Head and Neck Cancer Cells are efficiently infected by Ad5/35 Hybrid Virus. Journal of Gene Medicine 8:1223-1231. Shinozaki, K., Suominen, E., Carrick, F., Sauter, B., Kähäri, V.-M., Lieber, A., Woo, S.L.C. and Savontaus, M. (2006). Efficient infection of endothelial cells by a capsid-modified adenovirus. Gene Therapy 13:52-59. Hutter, R., Valdiviezo, C., Sauter, B.V., Savontaus, M., Chereshnev, I., Carrick, F.E., Bauriedel, G., Luderitz, B., Fallon, J.T., Fuster, V. and Badimon, J.J. (2004) Caspase-3 and tissue factor expression in lipid-rich plaque macrophages: evidence for apoptosis as link between inflammation and atherothrombosis. Circulation 27;109(16):2001-8. Ebert, O., Shinozaki, K., Huang, T.-G., Savontaus, M., GarciaSastre, A. and Woo S.L.C. (2003) VSV as oncolytic virus for treatment of orthotopic hepatocellular carcinoma in immunecompetent rats. Cancer Research 63(13):3605-11. Huang, T.-G., Savontaus, M., Shinozaki, K., Sauter, B. and Woo, S.L.C. (2003) Telomerase dependent oncolytic adenovirus for cancer treatment. Gene Therapy 10(15):1241-7. Savontaus, M., Sauter, B.V., Huang, T.-G. and Woo, S.L.C. (2002) Transcriptional Targeting of conditionally Replicating Adenovirus to Dividing Endothelial Cells. Gene Therapy 9(14): 972-979 Currently the main focus of our group is in gene therapy for cardiovascular disease. We are building on our previous findings by analyzing the adenovirus receptor expression and vector transduction efficiency in samples from patients with ischemic or 80 81 TRANSCRIPTIONAL REGULATION OF HEAT SHOCK GENE EXPRESSION Principal Investigator: Lea Sistonen, Ph.D., Professor of Cell and Molecular Biology, Department of Biosciences, Åbo Akademi University. Laboratory address: Centre for Biotechnology, BioCity, Tykistökatu 6, P.O.BOX 123, FI-20521 Turku, Finland. Tel. +358-2-333 8028, 215 3311; Fax +358-2-333 8000; Email: [email protected], [email protected] Biography: Lea Sistonen (b. 1959) completed her undergraduate studies at Åbo Akademi University in 1984 and received her Ph.D. from the University of Helsinki in 1990. She was a post-doctoral fellow at Northwestern University in Dr. Richard I. Morimoto’s laboratory in 1990-1993 (Fogarty International Fellowship 1991-1993). In November 1993 she joined the Centre for Biotechnology as a senior research fellow in molecular biology. In April 2000 she was appointed as Professor of Cell and Molecular Biology at Åbo Akademi University. During the 5-year period 2004-2009 she was Academy Professor, the Academy of Finland. Personnel: Post-doctoral fellows: Julius Anckar, Ph.D., Eva Henriksson, Ph.D., Malin Åkerfelt, Ph.D. Graduate students: Johanna Ahlskog, M.Sc., Johanna Björk, M.Sc., Henri Blomster, M.Sc., Zhanna Chitikova, M.Sc., Alexandra Elsing, M.Sc., Anton Sandqvist, M.Sc., Anniina Vihervaara, M.Sc. Technician: Helena Saarento, M.Sc. Undergraduate students: Anna Aalto, Heidi Bergman, Malin Blom, Marek Budzynski, Henrica Karlberg, Karoliina Rautoma, Jenny Siimes, Aki Vartiainen Description of the Project: The heat shock response is an evolutionarily well-conserved cellular defence mechanism against protein-damaging stresses, such as elevated temperatures or hyperthermia, heavy metals, and viral and bacterial infections. The heat shock proteins (Hsps) function as molecular chaperones to protect cells by binding to partially denatured proteins, dissociating protein aggregates, and regulating the correct folding and intracellular translocation of newly synthesized polypeptides. Hsps are transcriptionally regulated by heat shock factors, HSFs. The mammalian HSF family consists of four members HSF1-4. Although HSFs are best known as inducible transcriptional regulators of genes encoding molecular chaperones and other stress proteins, they are also important for normal developmental processes and longevity pathways. The repertoire of HSF targets has recently expanded well beyond the heat shock genes, and the known functions governed by HSFs span from the heat shock response to development, metabolism, lifespan and disease, especially cancer and neurodegenerative disorders. Our main interest is in elucidating the molecular mechanisms by which the different members of the HSF family are regulated during normal development and under stressful conditions. In particular, we investigate both the expression and activity of HSF1 and HSF2. We have found that HSF1 is ubiquitously expressed and its activity is primarily regulated by various post-translational modifications 82 (PTMs), such as acetylation, phosphorylation and sumoylation. All these PTMs are induced by stress stimuli but their effects on HSF1 vary. While examining the multi-site phosphorylation of HSF1, we observed that in response to stress, HSF1 undergoes phosphorylation-dependent sumoylation within a bipartite motif which we found in many transcription factors and co-factors and gave name PDSM (phosphorylation-dependent sumoylation motif. Stress-inducible hyperphosphorylation and sumoylation of HSF1 occur very rapidly, whereas acetylation of HSF1 increases gradually, indicating a role for acetylation in the attenuation phase of the HSF1 activity cycle. Indeed, we have shown that among multiple lysine residues targeted by acetylation, K80 is located within the DNA-binding domain of HSF1 and its acetylation is required for reducing HSF1 DNA-binding activity. Moreover, the duration of HSF1 DNA-binding activity could be prolonged or diminished by chemical compounds either activating or inhibiting the activity of the longevity factor deacetylase SIRT1. These results suggest that SIRT1-mediated deacetylation of HSF1 could maintain HSF1 in a state competent for DNA-binding, thereby linking our research to HSF1-mediated regulation of lifespan. Currently, our focus is on a complex network of PTMs to decipher the post-translational signature of HSF1. Unlike HSF1, which is a stable protein evenly expressed in most tissues and cell types, HSF2 shows a highly specific spatiotemporal expression pattern during development, and we have demonstrated that the amount of HSF2 is directly linked to its activity. Using mouse spermatogenesis as a model system, we have discovered an inverse correlation between the cell- and stage-specific wavelike expression patterns of HSF2 and a specific microRNA, miR-18, which is a member of the Oncomir-1/miR-17∼92 cluster. Intriguingly, miR-18 was found to repress the expression of HSF2 by directly targeting its 3’UTR. To investigate the in vivo function of miR-18, we developed a novel method T-GIST (Transfection of Germ cells in Intact Seminiferous Tubules) and were able to show that inhibition of miR-18 in intact mouse seminiferous tubules leads to increased HSF2 protein levels and altered expression of HSF2 target genes, including the Y-chromosomal multi-copy genes that we previously have reported as novel HSF2 targets in the testis. Our original finding that miR-18 regulates HSF2 activity in spermatogenesis links miR-18 to HSF2-mediated physiological processes and opens a whole new window of opportunities to elucidate the physiological and stress-related functions of HSF2, either alone or in conjunction with HSF1. Our studies on the formation of heterotrimers between HSF1 and HSF2 and their impact on already established and newly discovered targets genes should also shed light on the roles of HSFs in protein-misfolding disorders, such as neurodegenerative diseases, as well as in aging and cancer progression. So far, the studies have mostly concentrated on HSF1, but it is important to consider the existence of multiple HSFs and interactions between them, especially when searching for potential drugs to modify either expression or activity of these multi-faceted transcriptional regulators. Funding: The Academy of Finland, the Sigrid Jusélius Foundation, the Finnish Cancer Organizations, and Åbo Akademi University (Centre of Excellence in Cell Stress). Collaborators: Elisabeth Christians (University of Toulouse, France), Sampsa Hautaniemi (University of Helsinki), Susumu Imanishi and John 83 Eriksson (Åbo Akademi University), Noora Kotaja and Jorma Toppari (University of Turku), Pia Roos-Mattjus, Tiina Salminen, Peter Slotte and Kid Törnquist (Åbo Akademi University), Valérie Mezger (University of Paris Diderot, France), Jorma Palvimo (University of Eastern Finland), Sandy Westerheide and Rick Morimoto (Northwestern University, USA). Selected Publications: Björk J.K.*, Sandqvist A.*, Elsing A.N., Kotaja N. and Sistonen L. (2010) miR-18, a member of OncomiR-1, targets heat shock transcription factor 2 in spermatogenesis. Development, in press. Åkerfelt M., Morimoto R.I. and Sistonen L. (2010) Heat shock factors: integrators of cell stress, development and lifespan. Nat. Rev. Mol. Cell Biol. 11: 545-555. Blomster H.A.*, Imanishi S.Y.*, Siimes J., Kastu J., Morrice N.A., Eriksson J.E. and Sistonen L. (2010) In vivo identification of sumoylation sites by a signature tag and cysteine-targeted affinity purification. J. Biol. Chem. 285: 19324-19329. Blomster H.A., Hietakangas V., Wu J., Kouvonen P., Hautaniemi S. and Sistonen L. (2009) Novel proteomics strategy brings insight into the prevalence of SUMO-2 target sites. Mol. Cell. Proteomics 8: 1382-1390. Westerheide S.D.*, Anckar J.*, Stevens S.M.Jr., Sistonen L. and Morimoto R.I. (2009) Stress-inducible regulation of heat shock factor 1 by the deacetylase SIRT1. Science 323: 1063-1066. Sandqvist A., Björk J.K., Åkerfelt M., Chitikova Z., Grichine A., Vourc’h C., Jolly C., Salminen T.A., Nymalm Y. and Sistonen L. (2009) Heterotrimerization of heat-shock factors 1 and 2 provides a transcriptional switch in response to distinct stimuli. Mol. Biol. Cell 20: 1340-1347. Åkerfelt M.*, Henriksson E.*, Laiho A., Vihervaara A., Rautoma K., Kotaja N. and Sistonen L. (2008) Promoter ChIP-chip analysis in mouse testis reveals Y chromosome occupancy by HSF2. Proc. Natl. Acad. Sci. USA 105: 11224-11229. Östling P.*, Björk J.K.*, Roos-Mattjus P., Mezger V. and Sistonen L. (2007) HSF2 contributes to inducible expression of hsp genes through interplay with HSF1. J. Biol. Chem. 282: 7077-7086. Chang Y.*, Östling P.*, Åkerfelt M., Trouillet D., Rallu M., Gitton Y., El Fatimy R., Fardeau V., Le Crom S., Morange M., Sistonen L. and Mezger V. (2006) Role of heat shock factor 2 in cerebral cortex formation and as a regulator of p35 expression. Genes Dev. 20: 836-847. From left to right, standing: Lea Sistonen, Eva Henriksson, Johanna Björk, Jenny Siimes, Malin Åkerfelt, Jenni Vasara, Alexsandra Elsing, Malin Blom, Aki Vartiainen, sitting: Johanna Ahlskog, Anton Sandqvist, Mikael Puustinen, Marek Budzynski. Anckar J.*, Hietakangas V.*, Denessiouk K., Thiele D.J., Johnson M.S. and Sistonen L. (2006) Inhibition of DNA binding by differential sumoylation of heat shock factors. Mol. Cell. Biol. 26: 955-964. Hietakangas V.*, Anckar J.*, Blomster H.A., Fujimoto M., Palvimo J.J., Nakai A. and Sistonen L. (2006) PDSM, a motif for phosphorylation-dependent SUMO modification. Proc. Natl. Acad. Sci. USA 103: 45-50 (epub. Dec 21, 2005). *equal contribution 84 85 Cancer Cell Signaling http: http://www.btk.fi/index.php?id=1279 Principal investigator: Jukka Westermarck, M.D., Ph.D., Docent in Molecular Biology (University of Turku). Address: Turku Centre for Biotechnology, BioCity, Tykistökatu 6 B, P.O. Box 123, FIN-20251 Turku, Finland. Tel. +358-2-333 8621, Fax +358-2-333 8000. Email: [email protected], Biography: Jukka Westermarck (b. 1969) received his M.D. in 1996 and Ph.D in 1998 at the University of Turku. He was a postdoctoral fellow at European Molecular Biology Laboratory in Heidelberg, Germany, in Dr. Dirk Bohmann´s laboratory during 1999-2001. He was a Academy of Finland senior scientist during 2002-2007 and 20062009 he was appointed as a Group leader at Institute of Medical Technology (IMT), University of Tampere, Finland. In 2008 he was appointed to a Research Professor position at the Finnish Cancer Institute. 2009 he was appointed to Research director position at Turku Centre for Biotechnology (leave of absence until 2011). Personnel: Seniors scientist: Jukka Westermarck, M.D., Ph.D. Post-doctoral researchers: Christophe Come, Ph.D., Juha Okkeri, Ph.D., Yuba Pokharel, Ph.D., Sami Ventelä, M.D., Ph.D. Graduate students: Antoine Mialon, M.Sc., Minna Niemelä, M.Sc., Anni Laine, M.Sc., Tuuli Halonen, M.Sc., Amanpreet Kaur, M.Sc., Anchit Khanna, M.Sc. (IMT) Technical personnel: Taina Kalevo-Mattila Description of the project : The goal of our research group is to identify novel signaling mechanisms involved in malignant cell growth by isolating protein complexes associated with proteins previously demonstrated to have an important role in cancer progression. To identify protein complexes, we use tandem affinity purification (TAP) and Streptag purification methods, both proven to be suitable for purification of signaling protein complexes from mammalian cells in culture. Identification of novel proteins involved in malignant growth may also reveal novel possibilities for intervention in the therapy of cancer and other hyperproliferative diseases. Based on our recent work, we have identified several novel interacting proteins for signaling proteins such as AP-1 transcription factor c-Jun, MAPK kinase MEK1, and protein phosphatase PP2A. Most of our future work will be focused on characterization of PP2A interaction partner CIP2A, that we have demonstrated to inhibit PP2A in human malignancies. As PP2A inhibition has been recognized as a prerequisite for human cell transformation, it is plausible that further understanding of the function of CIP2A will reveal fundamental novel information about the basic mechanisms of cancer progression. The overall goal of the proposed project is to study the function and importance of CIP2A in cancer progression by using combination of molecular biology, cell biology and functional genetics methods. As our current results suggest that targeting CIP2A could be beneficial in the treatment of cancer, our goal is also to develop research models for evaluating the suitability of CIP2A as a novel drug target for cancer therapies. In addition, our aim is to purify new protein complexes related cancer cell signaling. 86 Funding: The Academy of Finland, Medical Research Fund of Tampere University Hospital, Turku Graduate School of Biomedical Sciences, Tampere Graduate School in Biomedicine and Biotechnology, Emil Aaltonen Foundation, Sigrid Juselius Foundation, Cancer Research Foundation of Finland, Association of International Cancer Research (UK). Collaborators: Tuula Kallunki (Danish Cancer Society), Rosalie Sears (Oregon Health and Science University), Owen Sansom (Beatson Institute for Cancer Research, Glasgow), Kirmo Wartiovaara (University of Helsinki), Sampsa Hautaniemi (University of Helsinki), Ari Ristimäki (University of Oulu), Jorma Toppari (University of Turku), Veli-Matti Kähäri (Turku University Hospital), Reidar Grenman (Turku University Hospital). Selected Publications: Kerosuo L, Fox H, Perälä N, Ahlqvist K, Suomalainen A, Westermarck J, Sariola H, and Wartiovaara K; CIP2A increases self-renewal and is linked to Myc in neural progenitor cells. Differentiation, in press, 2010 Heikkinen PT, Nummela M, Leivonen SK, Westermarck J, Hill CS, Kähäri VM, and Jaakkola PM; Hypoxia activated Smad3-specific dephosphorylation by PP2A. The Journal of Biological Chemistry, 285, 3740-3749, 2010. Come C, Laine A, Chanrion M, Edgren H, Mattila E, Liu X, Jonkers J, Ivaska J, Isola J, Darbon J-M, Kallioniemi O-P, and Thezenas S and Westermarck J; CIP2A is associated with human breast cancer aggressivity. Clinical Cancer Research, 15, 5092-5100, 2009. Khanna A, Böckelman C, Hemmes A, Junttila MR, Wiksten J-P, Lundin P, Junnila S, Murphy D, Evan GI, Haglund C, Westermarck J*, and Ristimäki A*; c-Myc-dependent regulation and prognostic role of CIP2A in gastric cancer. Journal of the National Cancer Institute, 101, 793-805, 2009. *equal contribution Puustinen P, Junttila MR, Vanhatupa S, Sablina AA, Hector ME, Teittinen K, Raheem O, Ketola K, Lin S, Kast J, Haapasalo H, Hahn WC, and Westermarck J; PME-1 Protects ERK Pathway activity from Protein Phosphatase 2A-mediated Inactivation in human malignant glioma. Cancer Research, 69, 2870-2877, 2009. Wu J, Ovaska K, Vallenius T, Westermarck J, Mäkelä TP, and Hautaniemi S; Protein-protein interaction portal for network level analysis. Nature Methods, 6, 75-77, 2009 Westermarck J, Hahn WC; Multiple pathways regulated by the tumor suppressor PP2A in transformation. Trends in Molecular Medicine, 14,152-160, 2008. Junttila MR, Li S-P, Westermarck J; Phosphatase-mediated crosstalk between MAPK signaling pathways in the regulation of cell survival. The FASEB Journal, 22, 954-965, 2008. Holmström TH, Mialon A, Kallio M, Nymalm Y, Mannermaa L, Holm T, Johansson H, Black E, Gillespie DA, Salminen TA, Langel U, Valdez BC, and Westermarck J; c-Jun supports ribosomal RNA processing and nucleolar localization of a RNA helicase DDX21. 87 The Journal of Biological Chemistry, 283, 7046-7053, 2008. Junttila, MR, Puustinen P, Niemelä M, Ahola R, Arnold H, Böttzauw T, Ala-aho R, Nielsen C, Ivaska J, Taya Y, Lu SL, Li S, Chan EKL, Wang X-J, Grenman R, Kast J, Kallunki T, Sears R, Kähäri V-M, Westermarck J; CIP2A Inhibits PP2A in Human Malignancies. Cell, 130, 51–62, 2007. Junttila MR, Ala-aho R, Jokilehto T, Peltonen J, Grenman R, Jaakkola P, Westermarck J, and Kähäri V-M; p38alpha and p38delta mitogen-activated protein kinase isoforms regulate invasion and growth of head and neck squamous carcinoma cells. Oncogene, 26, 5267-5279, 2007. Turku Centre for Biotechnology Ph.D. Theses 2009 1. Mialon, Antoine: Role and function of c-Jun protein complex in cancer cell behavior. University of Turku, p. 120 2. Pellinen, Teijo: Beta1 integrin regulation. University of Turku, p. 109 3. Mattila, Elina: Negative regulation of receptor tyrosine kinases by T-cell protein tyrosine phosphatase. University of Turku, p. 126. 4. Anckar, Julius: Multisite post-translational regulation of heat shock transcription factors. Åbo Akademi University, p. 162. 5. Kochin, Vitaly: Finding and characterizing protein phosphorylation sites that determine cellular decisions and functions. Åbo Akademi University, p. 185. 6. Kaunisto, Aura: Differential regulation of c-FLIP isoforms through post-translation al modifications. University of Turku, p.138. 7. Åkerfelt, Malin: Novel target genes for heat shock factors 1 and 2 in development. Åbo Akademi University, p. 166. Publications 2009 1. Aflakian, N., Ravichandran, S., Sarwar Jamaal, Md. S., Järvenpää, H., Lahesmaa, R. & Rao, K.V.S. 2009. Integration of signals from the B-cell antigen receptor and the IL-4 receptor leads to a cooperative shift in the cellular response axis. Mol. Biosyst., 5:1661-1671. 2. Ahonen, L.J., Kukkonen, A.M., Pouwels, J., Bolton, M.A., Jingle, C.D., Stukenberg, P.T. & Kallio, M.J. 2009. Perturbation of Incenp function impedes anaphase chromatid movements and chromosomal passenger protein flux at centromeres. Chromosoma, 118:71-84. 3. Axarli, I., Dhavala, P., Papageorgiou, A.C. & Labrou, N.E. 2009. Biochem. J., 422:247-256. 4. Axarli, I., Dhavala, P., Papageorgiou, A.C. & Labrou, N.E. 2009. J. Mol. Biol., 385:984-1002. 5. Blomster, H.A., Hietakangas, V., Wu, J., Kouvonen, P., Hautaniemi, S. & Sistonen, L. 2009. Mol. Cell. Proteomics, 8:1382-1390. 6. Brandt, D.T., Baarlink, C., Kitzing, T.M., Kremmer, E., Ivaska, J., Nollau, P. & Grosse, R. 2009. Nat. Cell Biol., 11:557-568. 7. Cheng, F., Weidner-Glunde, M., Varjosalo, M., Rainio, E.M., Lehtonen, A., Schulz, T.F., Koskinen, P.J., Taipale, J. & Ojala, P.M. 2009. From left to right, front row: Jukka Westermarck, Tiina Laiterä, Anna Cvrljevic, Leni Mannermaa, Christophe Côme, Yuba Raj Pokharel, Taina Kalevo-Mattila, Minna Niemelä, Tuuli Halonen, Kowstan Eskandari, Amanpreet Kaur, second row: Juha Okkeri, Anni Laine, Sami Ventelä, Otto Kauko. 88 PLoS Pathog., 5:e1000324. 8. Cho, S.H., Goenka, S., Henttinen, T., Gudapati, P., Reinikainen, A., Lahesmaa, R. & Boothby, M. 2009. PARP-14, a member of the B aggressive lymphoma (BAL) family, transduces survival 89 signals in primary B cells. Blood, 113:2416-2425. 9. Côme, C., Laine, A., Chanrion, M., Edgren, H., Mattila, E., Liu, X., Jonkers, J., Ivaska, J., Isola, J., Darbon, J.M., Kallioniemi, O., Thézenas, S. & Westermarck, J. 2009. Clin. Cancer Res., 15:5092-5100. 10. 11. Elo, L.L., Hiissa, J., Tuimala, J., Kallio, A., Korpelainen, E. & Aittokallio, T. 2009. Optimized detection of differential expression in global profiling experiments: case studies in clinical transcriptomic and quantitative proteomic datasets. Brief. Bioinform., 10:547-555. 12. Filén, J.J., Filén, S., Moulder, R., Tuomela, S., Ahlfors, H., West, A., Kouvonen, P., Kantola, S., Björkman, M., Katajamaa, M., Rasool, O., Nyman, T.A. & Lahesmaa, R. 2009. Quantitative proteomics reveals GIMAP family proteins 1 and 4 to be differentially regulated during human T helper cell differentiation. Mol. Cell. Proteomics, 8:32-44. 13. Hackauf, B., Rudd, S., van der Voort, J.R., Miedaner, T. & Wehling, P. 2009. Theor. Appl. Genet., 118:371-84. 14. Hiissa, J., Elo, L.L., Huhtinen, K., Perheentupa, A., Poutanen, M. & Aittokallio, T. 2009. Resampling reveals sample-level differential expression in clinical genome-wide studies. OMICS, 13:381-396. 15. Holmström, T.H., Rehnberg, J., Ahonen, L.J. & Kallio, M.J. 2009. Mol. Oncol., 3:262-268. 16. Iljin, K., Ketola, K., Vainio, P., Halonen, P., Kohonen, P. Clin. Cancer Res., 15:60706078. 17. Imanishi, S.Y., Kouvonen, P., Smått, J.H., Heikkilä, M., Peuhu, E., Mikhailov, A., Ritala, M., Lindén, M., Corthals, G.L. & Eriksson, J.E. 2009. 18. 19. 23. 24. 25. 26. 27. 28. 29. 30. 31. 2009. Neuropeptide Y polymorphism significantly magnifies diabetes and cardiovascular disease risk in obesity: the Hoorn Study. Eur. J. Clin. Nutr., 63:150-152. BMC Res. Notes, 2:204. 20. Kaunisto, A., Kochin, V., Asaoka, T., Mikhailov, A., Poukkula, M., Meinander, A. & Eriksson, J.E. 2009. Cell Death Differ., 16:1215-1226. 21. Khanna, A., Böckelman, C., Hemmes, A., Junttila, M.R., Wiksten, J.P., Lundin, M., Junnila, S., Murphy, D.J., Evan, 90 22. 32. 33. 34. 91 35. 41. 42. 36. 37. 38. 43. 44. 39. 45. 40. 46. 47. 48. 49. 50. 51. 52. 53. 54. . 92 93 LIFE OUTSIDE THE LAB 94 95 96 TURUN BIOTEKNIIKAN KESKUS ÅBO BIOTEKNIKCENTRUM TURKU CENTRE FOR BIOTECHNOLOGY th 0 2 y p y p r a a s H nniver A Cell Signalling to Systems Biology Research TURKU CENTRE FOR BIOTECHNOLOGY REPORT 2009 TURUN BIOTEKNIIKAN KESKUS Tykistökatu 6 B P.O.BOX 123 FI 20521 Turku, Finland Tel: +358 2 333 8603, Fax 358 2 333 8000
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Turku Centre for Biotechnology Published by: Turku Centre for Biotechnology P.O. Box 123, FI-20521 Turku, Finland Tel. int. +358-2-333 8603, fax int. +358-2-333 8000 http://www.btk.fi Editorial Boa...
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