Abstract PR03: Linking AP/MS-driven protein-protein interaction networks and combination CRISPR/sgRNA screens defines new Kras effectors and target candidates for non-small cell lung cancer

The Kras pathway is a central driver in prevalent and deadly cancers including pancreas, colon and lung. Despite decades of work studying the Raf and PI3 kinase pathways downstream of Kras, therapies targeted to these pathways have shown varied responses in tumorsTo better understand whether important Kras dependences have been overlooked, we used tandem affinity purification of Kras, Hras, and Nras interactors and a set of baits representing ~50 interacting pathway regulators, and have compared these interactions in Kras transformed A549 NSCLC lines versus an isogenic shRNA Kras knockdown line, to identify key protein-protein interactions (PPIs) linked to Kras effector activity. These interactions were curated and compared to public PPI, genetic susceptibility, and other multiomics data to assemble a physical PPI map annotated with important functional determinants. This functional map was used to construct a library of 114 selected sgRNAs (10 guides per gene) and screened in a 1140 x 1140 dual sgRNA vector system to look for quantitative changes in single and dual genetic dependencies in A549 cell growth. We discovered a considerable number of new KRAS GTP-specific effectors important in NSCLC cancer, many with strong mutations patterns supported by TCGA data and validated by in vitro biochemical assay. We will present these data and additional validation studies on two pathways including the Radil-Rap1-Kif14 pathway for cell migration and an extensive interaction of the Kr...
Source: Molecular Cancer Therapeutics - Category: Cancer & Oncology Authors: Tags: New Technology and Bioinformatics: Oral Presentations - Proffered Abstracts Source Type: research