Abstract A36: Genome-scale ORF screening to identify modulators of the epithelial-to-mesenchymal transition

Epithelial-mesenchymal transition (EMT) is an essential developmental program and is often reactivated during tumor initiation and progression. EMT is a reversible reprogramming of the cells. It not only promotes cell morphology alterations, but also provokes a profound cell state change in multiple regulatory circuits in global cell signaling, transcriptional and post-transcriptional modifications. Among all breast cancer subtypes, basal-like breast cancer displayed a high degree of mesenchymal and stem-like cell properties. To systematically interrogate the modulators of epithelial-to-mesenchymal transition, we performed a genome-scale ORF screen in human mammary epithelial cells. We used CD44 cell surface protein as a marker for cells reside in mesenchymal and stem-like cell state. CD44 low cells were presorted and induced with a barcoded genome-scale ORF library. After seven days of propagation, cells converted to CD44 high state were sorted by flow cytometry, and the barcodes enriched in the CD44 high population compared to the cells in the CD44 low population were evaluated with next-generation sequencing. Interestingly, 68 genes scored 3 standard deviations above the mean, including transcriptional factors, RNA splicing factors, epigenetic regulators, kinases/phosphatases, spermatogenesis regulators and amide metabolic modifiers. Strikingly, gene ontology and gene set enrichment analysis showed that RNA splicing process was the most significantly enriched biological pr...
Source: Molecular Cancer Therapeutics - Category: Cancer & Oncology Authors: Tags: Finding Synthetic Lethal Interactions through Functional Genomics: Poster Presentations - Proffered Abstracts Source Type: research