Identification of regulators of poly-ADP-ribose polymerase inhibitor response through complementary CRISPR knockout and activation screens

被引：38

作者：

Clements, Kristen E. ^{[1
]}

Schleicher, Emily M. ^{[1
]}

Thakar, Tanay ^{[1
]}

Hale, Anastasia ^{[1
]}

Dhoonmoon, Ashna ^{[1
]}

Tolman, Nathanial J. ^{[1
]}

Sharma, Anchal ^{[2
]}

Liang, Xinwen ^{[3
]}

Kawasawa, Yuka Imamura ^{[1
,4
,5
]}

Nicolae, Claudia M. ^{[1
]}

Wang, Hong-Gang ^{[3
,4
]}

De, Subhajyoti ^{[2
]}

Moldovan, George-Lucian ^{[1
]}

机构：

[1] Penn State Univ, Dept Biochem & Mol Biol, Coll Med, Hershey, PA 17033 USA

[2] Rutgers State Univ, Rutgers Canc Inst New Jersey, New Brunswick, NJ 08901 USA

[3] Penn State Univ, Dept Pediat, Coll Med, Hershey, PA 17033 USA

[4] Penn State Univ, Dept Pharmacol, Coll Med, Hershey, PA 17033 USA

[5] Penn State Univ, Inst Personalized Med, Coll Med, Hershey, PA 17033 USA

来源：

NATURE COMMUNICATIONS | 2020年 / 11卷 / 01期

关键词：

CYTOMETRY-BASED METHOD; HOMOLOGOUS RECOMBINATION; DAMAGED-CHROMATIN; END RESECTION; MUTANT-CELLS; 53BP1; ACETYLATION; POLY(ADP-RIBOSE); RESISTANCE; TIP60;

D O I：

10.1038/s41467-020-19961-w

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Inhibitors of poly-ADP-ribose polymerase 1 (PARPi) are highly effective in killing cells deficient in homologous recombination (HR); thus, PARPi have been clinically utilized to successfully treat BRCA2-mutant tumors. However, positive response to PARPi is not universal, even among patients with HR-deficiency. Here, we present the results of genome-wide CRISPR knockout and activation screens which reveal genetic determinants of PARPi response in wildtype or BRCA2-knockout cells. Strikingly, we report that depletion of the ubiquitin ligase HUWE1, or the histone acetyltransferase KAT5, top hits from our screens, robustly reverses the PARPi sensitivity caused by BRCA2-deficiency. We identify distinct mechanisms of resistance, in which HUWE1 loss increases RAD51 levels to partially restore HR, whereas KAT5 depletion rewires double strand break repair by promoting 53BP1 binding to double-strand breaks. Our work provides a comprehensive set of putative biomarkers that advance understanding of PARPi response, and identifies novel pathways of PARPi resistance in BRCA2-deficient cells.

引用

页数：14

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