The development of resistance to chemotherapies represents a significant barrier to successful cancer treatment. Resistance mechanisms are complex, can involve diverse and often unexpected cellular processes, and can vary with both the underlying genetic lesion and the origin or type of tumor. For these reasons developing experimental strategies that could be used to understand, identify and predict mechanisms of resistance in different malignant cells would be a major advance. Here we describe a novel gain-of-function forward genetic approach for identifying potential mechanisms of resistance. This system uses a modified piggyBac transposon to generate libraries of mutagenized cells, each containing transposon insertions that randomly activates nearby gene expression. By combining this with high-throughput sequencing, we have performed genome-wide screens for paclitaxel resistance mechanism in a variety of cancer cell lines. Validating this approach, we identified the multidrug transporter ABCB1 as the major contributor to resistance. In addition, we identified several potential novel resistance genes. Therefore we have developed a powerful new pipeline tool to systematically discover common and tumor specific pathways of resistance in human cancers. This cost-effective approach can be readily applied to different cell lines. Moreover, its ability to identify gene interactions and to probe complex genetic context provides a significant advantage over RNAi- or cDNA-based screens.
Citation Information: Mol Cancer Ther 2013;12(11 Suppl):C201.
Citation Format: Li Chen, Anahita Dastur, Cyril Benes. Transposon activation mutagenesis as a screening tool for identifying resistance to cancer therapeutics. [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2013 Oct 19-23; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2013;12(11 Suppl):Abstract nr C201.
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