期刊
MOLECULAR CELL
卷 68, 期 1, 页码 210-+出版社
CELL PRESS
DOI: 10.1016/j.molcel.2017.09.012
关键词
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资金
- NIH [P50 GM102706, U01 CA168370, R01 DA036858]
- post-doctoral fellowship [F32 GM116331]
- Swiss National Science Foundation [31003A_166608]
- European Research Council [ERC-STG 677936-RNAREG]
- Dutch Cancer Society (KWF)
- NWO CW ECHO [711.015.005]
- NIH/NCI [K99 CA204602, K99 CA181494]
- Stand Up To Cancer Innovative Research Grant
Chemical libraries paired with phenotypic screens can now readily identify compounds with therapeutic potential. A central limitation to exploiting these compounds, however, has been in identifying their relevant cellular targets. Here, we present a two-tiered CRISPR-mediated chemical-genetic strategy for target identification: combined genome-wide knockdown and overexpression screening as well as focused, comparative chemical-genetic profiling. Application of these strategies to rigosertib, a drug in phase 3 clinical trials for high-risk myelodysplastic syndrome whose molecular target had remained controversial, pointed singularly to microtubules as rigosertib's target. We showed that rigosertib indeed directly binds to and destabilizes microtubules using cell biological, in vitro, and structural approaches. Finally, expression of tubulin with a structure-guided mutation in the rigosertib-binding pocket conferred resistance to rigosertib, establishing that rigosertib kills cancer cells by destabilizing microtubules. These results demonstrate the power of our chemical-genetic screening strategies for pinpointing the physiologically relevant targets of chemical agents.
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