Systematic shRNA screens for the analysis of critical signaling pathways in KRAS-mutant multiple myeloma

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Mutations in KRAS are driver mutations in multiple myeloma (MM) yet no small moleculesthat target the encoded protein (KRAS) are available. Mutant KRAS activates multiple signal transduction pathways that act in unknown and potentially highly cell-specific combinations to drive growth of MM cells. In the previous funding period, we have established systematic loss-of-function (shRNA) screens as lentiviral pool screens coupled with next generation sequencing to a complexity of 10,000 shRNAs. We found that these screens can identify therapeutically relevant co-operations between downstream effector pathways of mutant KRAS. The applied-for project aims to firmly establish and validate this concept using different MM cell lines. Comparison with genome sequence data will establish whether specific biomarkers (for example mutations) reliably indicate co-operativity between critical signaling pathways. In parallel, we propose to design and clone a shRNA library that targets critical effector pathways with limited complexity in order to allow the analysis of KRASdependent signaling pathways in small cell populations (for example, in in vivo models of tumorigenesis).