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L1000 Dataset -small molecule, CRISPR perturbagens- LINCS Phase 2 (March 2017)

ABSTRACT: Transcriptional profiles of multiple cell and perturbation types: cells are treated with chemical perturbagens and CRISPR reagents. The expression level for 978 representative genes is measured.

PROVIDER: LDS-1372 | LINCS | 2016-03-06

REPOSITORIES: LINCS

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			Action	DRS
	BroadT_LINCS&file=LDS-1372.tar.gz	Other
	BroadT_LINCS&file=LDS-1482.tar.gz	Other
	BroadT_LINCS&file=LDS-1483.tar.gz	Other
	BroadT_LINCS&file=LDS-1484.tar.gz	Other

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Project description:High-throughput phenotype-based screening of large libraries of novel compounds without known targets can identify small molecules that elicit a desired cellular response, but additional approaches are required to find and characterize their targets and mechanisms of action. Here we show that a compound termed lung cancer screen 3 (LCS3), previously selected for its ability to impair the growth of human lung adenocarcinoma (LUAD) cell lines, but not normal lung cells, induces oxidative stress and activates the NRF2 signaling pathway by generating reactive oxygen species (ROS) in sensitive LUAD cell lines. To identify the target that mediates this effect, we applied thermal proteome profiling (TPP) and uncovered the disulfide reductases GSR and TXNRD1 as LCS3 targets. Through enzymatic assays using purified protein, we confirmed that LCS3 inhibits disulfide reductase activity through a reversible, uncompetitive mechanism. Further, we demonstrate that LCS3-sensitive LUAD cells are correspondingly sensitive to the synergistic inhibition of glutathione and thioredoxin pathways. Lastly, a genome-wide CRISPR knockout screen identified the loss of NQO1 as a mechanism of LCS3 resistance. This work highlights the ability of TPP to uncover targets of small molecules identified by high-throughput screens and demonstrates the potential utility of inhibiting disulfide reductases as a therapeutic strategy for LUAD.

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