Abstract 2537: CDK4/6 inhibition synergizes with KRAS-MAPK pathway targeting in pancreatic cancer

Abstract

Abstract Introduction: Pancreatic ductal adenocarcinoma (PDAC) is one of the most difficult human malignancies to treat due to its innate and acquired therapeutic resistance. Our hypothesis is that cyclin-dependent kinase 4 (CDK4) mediates therapeutic resistance to targeting of the KRAS-MAPK pathway in PDAC. Experimental Procedure: We characterized the expression of total and phosphorylated Retinoblastoma (Rb) and MEK protein levels in KRAS wild-type (BxPC3) and KRAS mutant (PANC1, MiaPaca2) human PDAC cell lines at baseline and with MEK and CDK4 inhibition alone or in combination. We then assessed the effects of combined therapy on cell-cycle progression and tumorigenicity (in vitro and in vivo). Finally, using the Ptf1a cre/+;LSL-Kras G12D/+;Tgfbr2 flox/flox (PKT) mouse model of PDAC, we assessed the in vivo overall tumor growth and survival after combined treatment with CDK4/6 and MEK inhibitors. Results: Rb functions as a tumor suppressor, and it is inactivated when phosphorylated by CDK4-Cyclin-D1. The CDK4/6 inhibitor (LEE011) effectively inhibits phosphorylation of Rb in cell lines regardless of KRAS mutational status. Combined inhibition of CDK4/6 and MEK (MEK162) decreased phosphorylation of RB and MAPK expression synergistically in the KRAS mutant cell lines, but not the KRAS wild-type BxPC3 cell line. Cell cycle progression was delayed effectively with MEK inhibition alone in the KRAS wild-type cell line, yet only combined CDK4/6 and MEK inhibition effectively delayed cell cycle progression in the KRAS mutant cell lines. Colony formation and invasion were also significantly decreased when Kras mutant cells were treated with combined CDK4/6 and MEK inhibition compared to all monotherapy and control groups. Finally, treatment of PKT mice resulted in a modest increase in OS with MEK inhibition alone, but mice receiving combined CDK4/6 and MEK inhibition exhibited a four-fold increase in OS. Conclusions: Combined inhibition of CDK4/6 and MEK results in significantly enhanced therapeutic efficacy and prolonged survival in the aggressive PKT mouse model of PDAC. This study suggests that concurrent inhibition of CDK4/6 and MEK may be an effective treatment for PDAC. Citation Format: Jason A. Castellanos, Nagaraj Nagathihalli, Michael N. Van Saun, Cameron Kasmai, Yanhua Xiong, Nipun Merchant. CDK4/6 inhibition synergizes with KRAS-MAPK pathway targeting in pancreatic cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 2537. doi:10.1158/1538-7445.AM2015-2537