Abstract While mutant (mt) KRas has proven to be difficult to target directly, mt KRas-driven cancer cells may gain dependencies on other signal transduction pathways. Therefore, exploring these dependencies may lead to the discovery of pathways and targets whose inhibition may induce synthetic lethality in cancer cells that depend on mt KRas. We have screened a kinase inhibitor library against 7 human cancer cell lines that harbor mt KRas; 4 mt KRas-dependent (A427 and H358 (lung), SW620 and SK-CO-1 (colon)); and 3 mt KRas-independent (HCT116, DLD-1 and HCT-8 (colon)) to identify kinase inhibitors that selectively inhibit the survival of mt KRas-dependent cancer cells. Our results show that 13 of the kinase inhibitors suppressed potently the survival of all 7 human cancer cell lines, 31 inhibited very weakly and 84 inhibited differentially each cell line, and the remaining compounds showed varied activities across cell lines. Importantly, 7 kinase inhibitors inhibited preferentially (p< 0.01) the viability of mt KRas-dependent over -independent human cancer cell lines. These results suggested that the 7 compounds may target kinases whose inhibition preferentially affects mt KRas-dependent cells. Analysis of the kinases that are targeted by these inhibitors revealed that these compounds target predominantly tyrosine kinases suggesting that at least in these cell lines these compounds may disrupt mt KRas “addiction” by inhibiting tyrosine kinase-dependent signaling. Citation Format: Norbert Berndt, Xiaolei Zhang, Rays H.Y. Jiang, Said M. Sebti. Kinome inhibitor screen identifies kinase inhibitors that inhibit selectively the survival of mutant K-Ras-dependent human cancer cells. [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 2153. doi:10.1158/1538-7445.AM2015-2153