Animal Models Ad Advances in Endometrial Cancer Epidemiology and Biology i E d i l C E id i l d Bi l Harvard School of Public Health 2014 Diego H. Castrillón, M.D., Ph.D. Department of Pathology and Department of Pathology and Simmons Comprehensive Cancer Center University of Texas Southwestern Medical Center D ll Dallas, Texas T
DISCLOSURES • Consultant / Scientific Advisor / Inventor on IP licensed to / / dasdf dasdf Molecular MD (LKB1 and oncology biomarker diagnostics)
Genetically-engineered mouse models of human cancer d l f h • Cancer is defined by complex tumor/host cell interactions y 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 Human = “simplex type” uterus Mouse = “duplex type” uterus p yp
endometrial-specific gene endometrial specific gene Discovery of Sprr2f as 1000 2000 3000 4000 5000 0.8 - 0 0.8 - Neonatal Ovary Adult Ovary U t e 1 2 3 4 5 Eggs r Cycle Day: T u e s Uterus s Cumulus Cells t i s S k ES Cells i n T Testis o n g E11 Embryo L u e u n Adrenal Gland O v a r y g H e Placenta a (Estrus) ( s us) MG r S t Day 4 k Uterus e Uterus l S e t B. Marrow p a Vagina l e l M e T Spleen n h u y s s + - 3 Wk c c m Thymus I E2 l e n u t s e Brain s L t i i n v Eye e e r S t Sk. Muscle o m K Heart a i c d h n Intestine B e r y a Kidney i n Liver Lung
Sprr2f RNA ISH Sprr2f RNA ISH In situ Endometrinin Contreras et al Dis Model Mech 2010 3:181
Generation and validation of Sprr2f-Cre Tg allele f S 2f C T ll l S Sprr2f-Cre 2f C x R26R x R26R Contreras et al Dis Model Mech 2010 3:181
Stereotypical tumor progression in Sprr2f-Cre; Lkb1 L/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 0.11g 0.19g 0.46g 1.17g 0.14g * * * * * * * cervix cervix bladder bladder extra uterine fat extra-uterine fat peritoneum peritoneum ovary o ar *
100 Cre; Lkb1 L/L (n=25) 80 Cre; Lkb1 L/+ (n=12) vival 60 +/+ (n=10) % surv 40 20 p < 0.0001 Sprr2f-Cre; Lkb1 L/L Lkb1 L/L S 2f C 0 0 0 200 400 600 800 1000 tumors are days 2.5 highly lethal highly lethal 2.0 weight (g) 1.5 wt 1.0 Cre; Lkb1 L/L Cre; Lkb1 0.5 0.0 0 6 4 2 0 8 6 1 4 6 9 3 5 8 1 1 1 1 days days
mTOR inhibitors for cancer therapy py Temsirolimus Rapamycin Everolimus (CCI-779) (RAD001) • Mixed results • Mixed results • No benefit: melanoma, GBM, etc. • Temsirolimus: FDA approved for RCC • T i li FDA d f RCC • Responses in endometrial ca • No predictive tests N di i
Rapamycin sensitivity in endometrial cancer cell lines endometrial cancer cell lines A B Cell line Rank Sensitivity (10 µM) AN3CA AN3CA 4 4 0 086 0.086 MFE-319 11 0.157 MFE-296 16 0.195 HEC-1 24 0.228 p=3x10 -5 Ishikawa 86 0.395 Heraklio 02 ER- 103 0.419 SNG-M 113 0.428 EN EN 149 149 0.474 0.474 EFE-184 155 0.48 MFE-280 374 0.635 C 28 f a particular tumor type e 24 20 16 f responding cell lines of 12 endometrial cancer 8 4 # of 0 0 10 20 30 40 50 60 70 80 90 100 Jeff Settleman % of responding cell lines of a particular tumor type
mTOR inhibitor trial: early disease y Sprr2f-Cre ; Lkb1 L/L ♀ ♀ Sprr2f Cre ; Lkb1 age: 12 weeks rapamycin rapamycin vehicle vehicle (2mg/kg qD) 4 weeks autopsy
1.0 p<0.0001 Lkb1 0.8 erine weight (g) 0.6 0.4 pa ute Lkb1+Rap 0.2 0.0 wt wt + Rapa Lkb1 Lkb1 + Rapa p=0.004 p<0.0001 Rapamycin halts 250 300 200 p=0.013 200 150 tumor progression positivity positivity ositivity 200 150 100 Ki67 po TUNEL p p-S6K p 100 100 50 50 0 0 0 wt Lkb1 Lkb1+Rapa wt Lkb1 Lkb1+Rapa wt Lkb1 Lkb1+Rapa p-S6K p wt Lkb1 Lkb1 + Rapa
mTOR inhibitor trial: advanced disease Sprr2f-Cre ; Lkb1 L/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 in tumor regression 1800 1800 ↓ 2.5 1500 † mm 3 ) 2.0 1200 volume (m ge fold chang 1.5 ↓ 900 † 1.0 ↓ ↓ 600 0.5 0.0 300 0 2 4 6 8 10 0 0 2 4 6 8 1 weeks weeks Pre-tx (0 wks) Pre tx (0 wks) 2 wks Rapa 2 wks Rapa 4 wks Rapa 4 wks Rapa 6 wks Rapa 6 wks Rapa 8 wks (2 wks off) 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) Lkb1 + oophorectomy (n=13) ival (%) 50 50 surv 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 6wks 12wks 20wks Lkb1 L/L Sprr2f-Cre; Lkb1 L/L 6 weeks 6 weeks 6 weeks 6 weeks Nakada et al PLOS ONE 2013 8:e73449
Telomere shortening in Type II cancers? TELO-CISH Type II << Type I 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 Pot1a / p53 tumors Akbay et al Oncogene 2013 32:2219
Summary/outlook Summary/outlook • Mouse models rich potential resource for understanding biology of endometrial carcinogenesis understanding biology of endometrial carcinogenesis (Type I vs. Type II) • Untapped potential in the ultimate goal of validation and individualization of cancer therapy and individualization of cancer therapy
Acknowledgments • Castrillon Lab • MGH Cancer Center Harvard – Cristina Contreras – Nabeel Bardeesy – Shana Wingo – Jeff Settleman – Esra Akbay*** – John Schorge – Teresa Gallardo Teresa Gallardo • DFCI – Marshall Haynie – Kwok Kin-Wong – George John – David Kwiatkowski – Meredith Shidler • Y l S h Yale School of Medicine l f M di i – Lane Shirley – Jose Michel – Sandy Chang – Yuji Nakada • UNC Lineberger – Chris Peña Chris Peña – Norman Sharpless Norman Sharpless – Dennis Ruder – Neil Hayes – Ileana Cuevas • MD Anderson – Russel Broaddus Russel Broaddus • Support • UTSW Advanced Imaging Center – NCI MMHCC UO1 – Masaya Takahashi – NCI RO1 – CPRIT Individual Investigator Award g – NIH Mouse Pathobiologist Award (K26) – Sidney Kimmel Foundation – ACS
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