dasdf

Animal Models
Ad
Advances
in
i Endometrial
E d
i lC
Cancer EEpidemiology
id i l
and
d Bi
Biology
l
Harvard School of Public Health 2014
Diego H. Castrillón, M.D., Ph.D.
Department of Pathology and
Simmons Comprehensive Cancer Center
University of Texas Southwestern Medical Center
D ll Texas
Dallas,
T
DISCLOSURES
• Consultant
/ Scientific Advisor / Inventor on IP licensed to
dasdf
Molecular MD (LKB1 and oncology biomarker diagnostics)
Genetically-engineered
mouse models
d l off human
h
cancer
• Cancer is defined byy complex
p tumor/host
/
cell interactions
• Cancer cannot be fully modelled in vitro
• Diversity of genetic tools and alleles is needed
• Useful for hypothesis testing and hypothesis generation
• Preclinical models / therapy individualization
• TCGA  many new endometrial cancer genes
The uterus: mouse vs. human
Human = “simplex type” uterus
Mouse = “duplex
p type”
yp uterus
Uterus
Cycle Day:
1 2 3 4 5
0.8 Ovary
MG
Uterus
Vagina
Day 4
((Estrus)
s us)
+
E2
Discovery of
Sprr2f as
endometrial specific gene
endometrial-specific
U
te
Te rus
s
Sk tis
i
To n
n
Lu gue
n
H g
ea
Sk rt
e
Sp let
l al
Th een Mu
sc
y
In mu
le
te s
s
Li tin
v e
St er
om
Ki ac
d h
Br n e y
ai
n
0.8 -
3 Wk
-
Lung
Liver
Kidney
Intestine
Heart
Sk. Muscle
Eye
Brain
Thymus
Spleen
B. Marrow
Uterus
Placenta
Adrenal Gland
E11 Embryo
Testis
ES Cells
Cumulus Cells
Eggs
Adult Ovary
Neonatal Ovary
5000
4000
3000
2000
1000
0
Sprr2f RNA ISH
In situ Endometrinin
Contreras et al
Dis Model Mech 2010 3:181
Generation and validation
off Sprr2f-Cre
S 2f C Tg
T allele
ll l
S 2f C
Sprr2f-Cre
x R26R
Contreras et al
Dis Model Mech 2010 3:181
Stereotypical tumor progression in
/ ♀
Sprr2f-Cre; Lkb1L/L
WT
16 weeks
Cre + Lkb1 L/L
6 weeks
0.11g
Cre + Lkb1 L/L
20 weeks
0.46g
1.17g
*
*
bladder
Cre + Lkb1 L/L
16 weeks
0.19g
0.14g
* *
cervix
Cre + Lkb1 L/L
12 weeks
o ar
ovary
extra uterine fat
extra-uterine
*
peritoneum
*
100
% surv
vival
Cre; Lkb1L/L (n=25)
80
Cre; Lkb1L/+ (n=12)
60
+/+
(n=10)
40
20
p < 0.0001
SSprr2f-Cre;
2f C Lkb1L/L
tumors are
highly lethal
0
0
200
400
600
800
1000
days
2.5
1.5
wt
1.0
L /L
Cre; Lkb1
0.5
days
19
6
16
8
14
0
11
2
84
56
0.0
30
weight (g)
2.0
mTOR inhibitors for cancer therapy
py
Rapamycin
•
•
•
•
•
Temsirolimus
Everolimus
(CCI-779)
(RAD001)
Mixed results
No benefit: melanoma, GBM, etc.
T
Temsirolimus:
i li
FDA approved
d ffor RCC
Responses in endometrial ca
N predictive
No
di i tests
Rapamycin sensitivity in
endometrial cancer cell lines
A
B
Cell line
Rank
Sensitivity (10 µM)
AN3CA
MFE-319
MFE-296
HEC-1
Ishikawa
Heraklio 02 ERSNG-M
EN
EFE-184
MFE-280
4
11
16
24
86
103
113
149
155
374
0 086
0.086
0.157
0.195
0.228
0.395
0.419
0.428
0.474
0.48
0.635
p=3x10
-5
C
# off responding cell lines off a particular tumor type
e
28
24
20
16
12
endometrial
cancer
8
4
0
0
10
20
30
40
50
60
70
80
% of responding cell lines of a particular tumor type
90
100
Jeff Settleman
mTOR inhibitor trial: earlyy disease
Sprr2f Cre; Lkb1L/L ♀
Sprr2f-Cre;
age: 12 weeks
rapamycin
(2mg/kg qD)
4 weeks
autopsy
vehicle
1.0
Lkb1
p<0.0001
ute
erine weight (g)
0.8
0.6
Lkb1+Rap
pa
0.4
0.2
0.0
wt
Lkb1
Lkb1 + Rapa
p=0.004
300
p<0.0001
250
200
200
100
0
200
150
p-S6K p
positivity
TUNEL p
positivity
Ki67 po
ositivity
p=0.013
100
50
Lkb1 Lkb1+Rapa
100
50
0
0
wt
150
wt
Lkb1 Lkb1+Rapa
wt
Lkb1 Lkb1+Rapa
p
p-S6K
Rapamycin halts
tumor progression
wt + Rapa
wt
Lkb1
Lkb1 + Rapa
mTOR inhibitor trial: advanced disease
Sprr2f-Cre;
p
; Lkb1L/L ♀
age >20 wks (advanced disease)
rapamycin
(2mg/kg qD)
serial
se
a MRI
M (q 2 wks)
w s)
Contreras et al
Dis Model Mech 2010 3:181
Rapamycin results in
tumor regression
1800
↓
2.5
†
2.0
900
fold chang
ge
1200
†
↓
600
1.5
1.0
↓
↓
0.5
0.0
10
8
6
4
2
300
0
volume (m
mm 3 )
1500
0
2
2 wks Rapa
4 wks Rapa
6
8
10
weeks
weeks
Pre-tx
Pre
tx (0 wks)
4
6 wks Rapa
8 wks (2 wks off)
Contreras et al
Dis Model Mech 2010 3:181
Oophorectomy prolongs survival in
Lkb1-driven endometrial cancer
100
Lkb1 (n=11)
survival (%)
Lkb1 + oophorectomy (n=13)
50
p = 0.0004
0
0
50
100
150
200
250
300
350
400
450
Age (days)
Dennis Ruder
Validation of Lkb1 mAB
(Cell Signalling clone #D60C5)
in mosaic mice (6 weeks)
100
80
60
40
20
0
3wks
Lkb1L/L
6 weeks
6wks
12wks
20wks
Sprr2f-Cre; Lkb1L/L
6 weeks
Nakada et al
PLOS ONE 2013 8:e73449
Telomere shortening in Type II cancers?
TELO-CISH
Type II << Type I
Akbay et al
Am J Pathol 2008 173:536
Endometrial cancer in Pot1a/p53 mice
Akbay et al
Oncogene 2013 32:2219
Type II features in Pot1a/p53
/
tumors
Akbay et al
Oncogene 2013 32:2219
Tetraploidy in
Pot1a/p53
tumors
Akbay et al
Oncogene 2013 32:2219
Summary/outlook
• Mouse models rich potential resource for
understanding biology of endometrial carcinogenesis
(Type I vs. Type II)
• Untapped potential in the ultimate goal of validation
and individualization of cancer therapy
Acknowledgments
•
•
Castrillon Lab
– Cristina Contreras
– Shana Wingo
– Esra Akbay***
– Teresa Gallardo
– Marshall Haynie
– George John
– Meredith Shidler
– Lane Shirley
– Jose Michel
– Yuji Nakada
– Chris Peña
– Dennis Ruder
– Ileana Cuevas
•
•
•
•
•
Support
•
– NCI MMHCC UO1
– NCI RO1
– CPRIT Individual Investigator
g
Award
– NIH Mouse Pathobiologist Award (K26)
– Sidney Kimmel Foundation
– ACS
MGH Cancer Center Harvard
– Nabeel Bardeesy
– Jeff Settleman
– John Schorge
DFCI
– Kwok Kin-Wong
– David Kwiatkowski
Y l School
Yale
S h l off Medicine
M di i
– Sandy Chang
UNC Lineberger
– Norman Sharpless
– Neil Hayes
MD Anderson
– Russel Broaddus
UTSW Advanced Imaging Center
– Masaya Takahashi