A Genome-wide CRISPR Screen Identifies CDC25A as a Determinant of Sensitivity to ATR Inhibitors

Sergio Ruiz; Cristina Mayor-Ruiz; Vanesa Lafarga; Matilde Murga; Maria Vega-Sendino; Sagrario Ortega; Oscar Fernandez-Capetillo

doi:10.1016/j.molcel.2016.03.006

A Genome-wide CRISPR Screen Identifies CDC25A as a Determinant of Sensitivity to ATR Inhibitors

Mol Cell. 2016 Apr 21;62(2):307-313. doi: 10.1016/j.molcel.2016.03.006. Epub 2016 Apr 7.

Authors

Sergio Ruiz¹, Cristina Mayor-Ruiz², Vanesa Lafarga², Matilde Murga², Maria Vega-Sendino², Sagrario Ortega³, Oscar Fernandez-Capetillo⁴

Affiliations

¹ Genomic Instability Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain. Electronic address: sruizm@cnio.es.
² Genomic Instability Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain.
³ Transgenics Unit, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain.
⁴ Genomic Instability Group, Spanish National Cancer Research Centre (CNIO), Madrid 28029, Spain; Science for Life Laboratory, Division of Translational Medicine and Chemical Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, 171 21 Stockholm, Sweden. Electronic address: ofernandez@cnio.es.

Abstract

One recurring theme in drug development is to exploit synthetic lethal properties as means to preferentially damage the DNA of cancer cells. We and others have previously developed inhibitors of the ATR kinase, shown to be particularly genotoxic for cells expressing certain oncogenes. In contrast, the mechanisms of resistance to ATR inhibitors remain unexplored. We report here on a genome-wide CRISPR-Cas9 screen that identified CDC25A as a major determinant of sensitivity to ATR inhibition. CDC25A-deficient cells resist high doses of ATR inhibitors, which we show is due to their failure to prematurely enter mitosis in response to the drugs. Forcing mitotic entry with WEE1 inhibitors restores the toxicity of ATR inhibitors in CDC25A-deficient cells. With ATR inhibitors now entering the clinic, our work provides a better understanding of the mechanisms by which these compounds kill cells and reveals genetic interactions that could be used for their rational use.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Antineoplastic Agents / pharmacology*
Ataxia Telangiectasia Mutated Proteins / antagonists & inhibitors
Ataxia Telangiectasia Mutated Proteins / genetics
Ataxia Telangiectasia Mutated Proteins / metabolism
CRISPR-Cas Systems*
Cell Cycle Proteins / antagonists & inhibitors
Cell Cycle Proteins / metabolism
Cell Line
Dose-Response Relationship, Drug
Drug Resistance, Neoplasm / genetics*
Embryonic Stem Cells / drug effects*
Embryonic Stem Cells / enzymology
Embryonic Stem Cells / pathology
Genome-Wide Association Study
Humans
Mitosis / drug effects
Molecular Targeted Therapy
Nuclear Proteins / antagonists & inhibitors
Nuclear Proteins / metabolism
Protein Kinase Inhibitors / pharmacology*
Protein-Tyrosine Kinases / antagonists & inhibitors
Protein-Tyrosine Kinases / metabolism
RNA Interference
Signal Transduction / drug effects
Transfection
cdc25 Phosphatases / genetics
cdc25 Phosphatases / metabolism*

Substances

Antineoplastic Agents
Cell Cycle Proteins
Nuclear Proteins
Protein Kinase Inhibitors
Atr protein, mouse
Protein-Tyrosine Kinases
Wee1 protein, mouse
Ataxia Telangiectasia Mutated Proteins
Cdc25a protein, mouse
cdc25 Phosphatases

Abstract

Publication types

MeSH terms

Substances

Grants and funding