Abstract C159: Identification of potential cancer drug targets using HT RNAi screening with pooled shRNA libraries.

Abstract

Genes modulating proliferation and survival in oncogenic cells have been identified using pooled lentiviral-based libraries expressing many thousands of shRNAs. Viability assays with human isogenic mammary epithelial cells (HMEC) and blood cancer cell lines transduced with these shRNA libraries targeting thousands of genes identified hundreds of essential genes for each panel of cells. Subsequent validation using single shRNA-expressing constructs showed that about 80% of the shRNAs identified in each complex library screen did in fact lead to cell death when transduced in cells. Analysis of the identified essential genes for known biological interactions revealed non-random clusters of interacting proteins that provide some insight into signaling pathways and protein networks specific to these cancers. Also, we have recently adapted the approach to combinatorially screen shRNA sequences targeting hundreds of genes to discover additive and synergistic combinations that generate a synthetic-lethal phenotype. Analysis of the lethal combinations indicates redundant, complementary, and compensatory responses in cancer cells. This presentation will provide an overview of the screening platform, our approach to shRNA library design, and results from the screens for cancer cell-specific lethal and synthetic-lethal genes. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C159.