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Animal ModelsAd i E d i l C E id i l d Bi lAdvances in Endometrial Cancer Epidemiology and Biology
Harvard School of Public Health 2014
Diego H. Castrillón, M.D., Ph.D.
Department of Pathology andDepartment of Pathology and Simmons Comprehensive Cancer Center
University of Texas Southwestern Medical CenterD ll TDallas, Texas
DISCLOSURES
dasdf• Consultant / Scientific Advisor / Inventor on IP licensed to dasdf / /Molecular MD (LKB1 and oncology biomarker diagnostics)
Genetically-engineeredd l f hmouse models of human cancer
• Cancer is defined by complex tumor/host cell interactionsy p /
• 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
Mouse = “duplex type” uterusHuman = “simplex type” uterus p yp
4000
5000
1000
2000
3000
scle
0
Neo
nata
lOva
ryAd
ultO
vary
Eggs
Cum
ulus
Cells
ESCe
lls
Test
isE1
1Em
bryo
Adre
nalG
land
Plac
enta
Uter
usB.
Mar
row
Sple
enTh
ymus
Bra
inEy
eSk
.Mus
cle
Hea
rtIn
test
ine
Kidn
eyLi
ver
Lung
Discovery of Sprr2f as
endometrial specific geneSkinTo
ngue
Lung
Heart
Skelet
al Mus
c
Spleen
Thymus
Intes
tine
Liver
Stomac
hKidn
eyBrai
n
Testi
s
Uterus
0.8 -endometrial-specific gene
Day 4(Estrus)
Ova
ryM
GU
teru
sVa
gina
E2
3 WkUterusCycle Day:
0.8 -
1 2 3 4 5
( s us)
+ -
Sprr2f RNA ISHSprr2f RNA ISH
In situ Endometrinin
Contreras et al Dis Model Mech 2010 3:181
Generation and validationf S 2f C T ll lof Sprr2f-Cre Tg allele
S 2f C
x R26R
Sprr2f-Cre
x R26R
Contreras et al Dis Model Mech 2010 3:181
Stereotypical tumor progression inSprr2f-Cre; Lkb1L/L ♀Sprr2f-Cre; Lkb1 / ♀
WT Cre + Lkb1 L/L Cre + Lkb1 L/L Cre + Lkb1 L/L Cre + Lkb1 L/L 16 weeks 6 weeks 12 weeks 16 weeks 20 weeks
0.46g0.19g0.14g 1.17g
* ***0.11g
bladder o arcervix extra uterine fat
* **
peritoneumbladder ovary cervix extra-uterine fat peritoneum
*
60
80
100Cre; Lkb1L/L (n=25)
+/+ (n=10)
Cre; Lkb1L/+ (n=12)
viva
l
S 2f C Lkb1L/L 0
20
40
p < 0.0001
% s
urv
Sprr2f-Cre; Lkb1L/L
tumors arehighly lethal 2.0
2.5
0 200 400 600 800 10000
days
highly lethal
1.0
1.5
wtCre; Lkb1L/Lw
eigh
t (g)
30 56 84 112
140
168
196
0.0
0.5Cre; Lkb1
daysdays
mTOR inhibitors for cancer therapypy
• Mixed results
Everolimus
(RAD001)
Temsirolimus
(CCI-779)
Rapamycin
• Mixed results• No benefit: melanoma, GBM, etc.• T i li FDA d f RCC• Temsirolimus: FDA approved for RCC• Responses in endometrial ca
N di i• No predictive tests
Rapamycin sensitivity inendometrial cancer cell linesA B
Cell line Rank Sensitivity (10 µM) AN3CA 4 0 086
endometrial cancer cell lines
AN3CA 4 0.086MFE-319 11 0.157 MFE-296 16 0.195 HEC-1 24 0.228 Ishikawa 86 0.395 Heraklio 02 ER- 103 0.419 SNG-M 113 0.428 EN 149 0.474
p=3x10-5
C 28
e
EN 149 0.474EFE-184 155 0.48 MFE-280 374 0.635
16
20
24
f a p
artic
ular
tum
or ty
pe
4
8
12
f res
pond
ing
cell
lines
of
endometrial cancer
00 10 20 30 40 50 60 70 80 90 100
% of responding cell lines of a particular tumor type
# of
Jeff Settleman
mTOR inhibitor trial: early diseasey
Sprr2f-Cre; Lkb1L/L ♀Sprr2f Cre; Lkb1 ♀age: 12 weeks
rapamycin vehiclerapamycin vehicle (2mg/kg qD)
4 weeks
autopsy
1.0 p<0.0001
0.4
0.6
0.8
erin
e w
eigh
t (g) Lk
b1pa
wt wt + Rapa Lkb1 Lkb1 + Rapa0.0
0.2ute
Lkb1
+Rap
150
200
250
posi
tivity
100
150
200
posi
tivity
p=0.013
200
300
ositi
vity
p=0.004 p<0.0001
Rapamycin haltstumor progression
wt Lkb1 Lkb1+Rapa0
50
100
p-S6
K p
wt Lkb1 Lkb1+Rapa0
50
TUNE
L p
wt Lkb1 Lkb1+Rapa0
100Ki
67 p
op-
S6K
p
wt Lkb1 Lkb1 + Rapa
mTOR inhibitor trial: advanced disease
Sprr2f-Cre; Lkb1L/L ♀p ; ♀age >20 wks (advanced disease)
rapamycin(2mg/kg qD)
serial MRI (q 2 wks)se a M (q w s)Contreras et al Dis Model Mech 2010 3:181
Rapamycin results intumor regression
1800
1200
1500
1800
†↓
mm
3 )
2.0
2.5
ge
600
900 †↓
volu
me
(m
0.5
1.0
1.5↓
↓fold
cha
ng
0 2 4 6 8 10
300
weeks
Pre-tx (0 wks) 2 wks Rapa 4 wks Rapa 6 wks Rapa 8 wks (2 wks off)
0 2 4 6 8 100.0
weeks
Pre tx (0 wks) 2 wks Rapa 4 wks Rapa 6 wks Rapa 8 wks (2 wks off)
Contreras et al Dis Model Mech 2010 3:181
Oophorectomy prolongs survival inLkb1-driven endometrial cancer
100
Lkb1 (n=11)Lkb1 + oophorectomy (n=13)
50ival
(%)
50
surv
0 50 100 150 200 250 300 350 400 4500
p = 0.0004
Age (days)
Dennis Ruder
Validation of Lkb1 mAB(Cell Signalling clone #D60C5)
in mosaic mice (6 weeks)
40
60
80
100
0
20
3wks 6wks 12wks 20wks
Lkb1L/L
6 weeksSprr2f-Cre; Lkb1L/L
6 weeks6 weeks 6 weeks
Nakada et alPLOS ONE 2013 8:e73449
Telomere shortening in Type II cancers?TELO-CISH
Type II << Type IAkbay et alAm J Pathol 2008 173:536
Type II << Type I
Endometrial cancer in Pot1a/p53 mice
Akbay et alOncogene 2013 32:2219
Type II features in Pot1a/p53 tumors/
Akbay et alOncogene 2013 32:2219
Tetraploidy in Pot1a/p53Pot1a/p53
tumors
Akbay et alOncogene 2013 32:2219
Summary/outlookSummary/outlook
• Mouse models rich potential resource for understanding biology of endometrial carcinogenesisunderstanding biology of endometrial carcinogenesis (Type I vs. Type II)
• Untapped potential in the ultimate goal of validation and individualization of cancer therapyand individualization of cancer therapy
Acknowledgments• Castrillon Lab • MGH Cancer Center Harvard
– Cristina Contreras– Shana Wingo– Esra Akbay***
Teresa Gallardo
– Nabeel Bardeesy– Jeff Settleman– John Schorge
– Teresa Gallardo– Marshall Haynie – George John – Meredith Shidler
• DFCI– Kwok Kin-Wong– David Kwiatkowski
Y l S h l f M di i– Lane Shirley– Jose Michel– Yuji Nakada– Chris Peña
• Yale School of Medicine– Sandy Chang
• UNC LinebergerNorman SharplessChris Peña
– Dennis Ruder– Ileana Cuevas
– Norman Sharpless– Neil Hayes
• MD Anderson – Russel Broaddus
• Support– NCI MMHCC UO1– NCI RO1– CPRIT Individual Investigator Award
Russel Broaddus• UTSW Advanced Imaging Center
– Masaya Takahashi
g– NIH Mouse Pathobiologist Award (K26)– Sidney Kimmel Foundation– ACS
