Genetically Engineered Murine Models of RCC William Y. Kim, M.D.
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1 Genetically Engineered Murine Models of RCC William Y. Kim, M.D. Co-director, Mouse Phase 1 Unit (MP1U) Departments of Medicine, Genetics, and Urology Lineberger Comprehensive Cancer Center University of North Carolina Chapel Hill, NC
2 ccrcc Genetically Engineered Murine Models: it took a while Circa 2001 Circa 2014
3 VHL associated Kidney Cancer: a model VHL +/- Renal Tubular Epithelial Cell VHL -/- Renal Cyst VHL -/- Renal Cell Carcinoma HIF? HIF * VHL VHL VHL Kim and Kaelin, Journal of Clinical Oncology, 2004 Hit 1 Hit 2 * Hit 3 * Oncogene activation or tumor suppressor gene loss
4 NextGen sequencing gives insights
5
6 Vhl inactivation is not sufficient for tumorigenesis Haase et al. PNAS, 2001 Rankin, Can Res, 2006 (Vhl L/L ) Ma, Can Res, 2003 (Vhl L/L ) Kleymenova, Carcinogenesis, 2004 (Vhl +/- ) Pritchett, Oncogene, 2014 (Vhl L/L ) Lee, Oncogene, 2009 (Vhl R167Q ) Frew, EMBO J, 2008 (Vhl; Pten) Lehmann, JASN, 2015 (Vhl L/L ; Kif3a L/L ) Wang, PNAS, 2014 (Vhl L/L ; Bap1 L/L ) Albers, EMBO Mol Med, 2013 (Vhl L/L ; Tp53 L/L ) Fu, Can Res, 2011 (HIF1a) Fu, Can Res, 2013 (HIF2a)
7 MYC is amplified and CDKN2A (INK4a/ARF) is lost in ccrcc Beroukhim et. al., Can Res, 2009 Sato et. al., Nat Gen, 2011 TCGA KIRC, Nat, 2013 VHL VHL VHL Myc Ink4a/Arf Ink4a/ARF Myc Ink4a/ARF Myc
8 Alleles Vhl - conditional knock-out Vhl KspCad-CreER T2 - tamoxifen-inducible expression Ksp CreER T2 Cdkn2a - germline knock-out 1b 1a 2 3 KspCad-rtTA conditional, doxycycline Ksp rtta Tet-O-MYC doxycycline dependent expression TetO MYC
9 Brave graduate student: Sean Bailey V (Vhl F/F ) M (TetO-Myc) I (Ink4a/Arf) Vhl Ksp rtta 1b 1a 2 3 Ksp CreER T2 TetO MYC
10 MYC has relevance to type II prcc
11 Overexpression of c-myc results in papillary-like RCC
12 Human prcc is enriched in gene signatures of MYC activation TCGA: KIRP
13 Growth M cell lines MYC dependent
14 H&E Vhl inactivation with c-myc overexpression results in kidney tumors with clear cell features
15 Growth VM and VIM cell lines are MYC dependent
16 VM and VIM cells have distinct gene expression patterns MYC inducible gene sets Cell cycle progression Ribosome biogenesis VIM-specific DNA methylation
17 VIM tumors are metastatic
18 VIM cells have higher levels of EMT and genes affecting ECM remodeling
19 VIM cells demonstrate increased matrigel invasion
20 Vhl deficient cells express Hif1alpha HIF2alpha?
21 pvhl restoration does not alter in vitro growth IB: actin IB: HA
22 Conclusions MYC is sufficient for kidney tumors that resemble prcc Vhl inactivation combined with MYC overexpression gives clear cell changes Vhl inactivation combined with MYC overexpression and Cdkn2a inactivation is sufficient for bona fide ccrcc with a subset of tumors developing liver metastases. Much work is left to be done.
23 Kim Lab Sean Bailey Jordan Kardos Sara Wobker Ryoichi Saito Tracy Rose Aleisha Smith Bhavani Krishnan Janet Leung Harper Wilson MP1U Dave Darr Chuck Perou Ned Sharpless Jose Roques Daniel Roth Kyle Stewart Michael Trinkler Volker Haase (Vanderbilt) Dorien Peters (Leiden University) Thomas Carroll (UTSW) Dean Felsher (Stanford) Katie Hoadley (UNC) Joel Parker (UNC) Matt Milowsky (UNC) UNC Urologic Oncology Raj Pruthi Urology Residents and Attendings Research Staff FUNDING SOURCES BCAN NIH/NCI Department of Defense Damon Runyon Foundation University Cancer Research Fund (UCRF) AACR Kure It American Cancer Society
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