A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor

Nicholas C Turner; Christopher J Lord; Elizabeth Iorns; Rachel Brough; Sally Swift; Richard Elliott; Sydonia Rayter; Andrew N Tutt; Alan Ashworth

doi:10.1038/emboj.2008.61

A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor

Nicholas C Turner, Christopher J Lord, Elizabeth Iorns, Rachel Brough, Sally Swift, Richard Elliott, Sydonia Rayter, Andrew N Tutt, Alan Ashworth

Research output: Contribution to journal › Article › peer-review

Abstract

Inhibitors of poly (ADP-ribose)-polymerase-1 (PARP) are highly lethal to cells with deficiencies in BRCA1, BRCA2 or other components of the homologous recombination pathway. This has led to PARP inhibitors entering clinical trials as a potential therapy for cancer in carriers of BRCA1 and BRCA2 mutations. To discover new determinants of sensitivity to these drugs, we performed a PARP-inhibitor synthetic lethal short interfering RNA (siRNA) screen. We identified a number of kinases whose silencing strongly sensitised to PARP inhibitor, including cyclin-dependent kinase 5 (CDK5), MAPK12, PLK3, PNKP, STK22c and STK36. How CDK5 silencing mediates sensitivity was investigated. Previously, CDK5 has been suggested to be active only in a neuronal context, but here we show that CDK5 is required in non-neuronal cells for the DNA-damage response and, in particular, intra-S and G(2)/M cell-cycle checkpoints. These results highlight the potential of synthetic lethal siRNA screens with chemical inhibitors to define new determinants of sensitivity and potential therapeutic targets.

Original language	English
Pages (from-to)	1368-77
Number of pages	10
Journal	EMBO Journal
Volume	27
Issue number	9
DOIs	https://doi.org/10.1038/emboj.2008.61
Publication status	Published - 7 May 2008

Keywords

Blotting, Western
Cell Cycle/drug effects
Cell Line, Tumor
Cyclin-Dependent Kinase 5/genetics
DNA Damage
DNA Repair
Flow Cytometry
Fluorescent Antibody Technique
HeLa Cells
Humans
Hydrogen Peroxide/pharmacology
Mitogen-Activated Protein Kinase 12/genetics
Poly(ADP-ribose) Polymerase Inhibitors
Poly(ADP-ribose) Polymerases/metabolism
Protein-Serine-Threonine Kinases/genetics
RNA, Small Interfering/genetics
Transfection

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10.1038/emboj.2008.61

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title = "A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor",

abstract = "Inhibitors of poly (ADP-ribose)-polymerase-1 (PARP) are highly lethal to cells with deficiencies in BRCA1, BRCA2 or other components of the homologous recombination pathway. This has led to PARP inhibitors entering clinical trials as a potential therapy for cancer in carriers of BRCA1 and BRCA2 mutations. To discover new determinants of sensitivity to these drugs, we performed a PARP-inhibitor synthetic lethal short interfering RNA (siRNA) screen. We identified a number of kinases whose silencing strongly sensitised to PARP inhibitor, including cyclin-dependent kinase 5 (CDK5), MAPK12, PLK3, PNKP, STK22c and STK36. How CDK5 silencing mediates sensitivity was investigated. Previously, CDK5 has been suggested to be active only in a neuronal context, but here we show that CDK5 is required in non-neuronal cells for the DNA-damage response and, in particular, intra-S and G(2)/M cell-cycle checkpoints. These results highlight the potential of synthetic lethal siRNA screens with chemical inhibitors to define new determinants of sensitivity and potential therapeutic targets.",

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author = "Turner, {Nicholas C} and Lord, {Christopher J} and Elizabeth Iorns and Rachel Brough and Sally Swift and Richard Elliott and Sydonia Rayter and Tutt, {Andrew N} and Alan Ashworth",

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AU - Turner, Nicholas C

AU - Lord, Christopher J

AU - Iorns, Elizabeth

AU - Brough, Rachel

AU - Swift, Sally

AU - Elliott, Richard

AU - Rayter, Sydonia

AU - Tutt, Andrew N

AU - Ashworth, Alan

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AB - Inhibitors of poly (ADP-ribose)-polymerase-1 (PARP) are highly lethal to cells with deficiencies in BRCA1, BRCA2 or other components of the homologous recombination pathway. This has led to PARP inhibitors entering clinical trials as a potential therapy for cancer in carriers of BRCA1 and BRCA2 mutations. To discover new determinants of sensitivity to these drugs, we performed a PARP-inhibitor synthetic lethal short interfering RNA (siRNA) screen. We identified a number of kinases whose silencing strongly sensitised to PARP inhibitor, including cyclin-dependent kinase 5 (CDK5), MAPK12, PLK3, PNKP, STK22c and STK36. How CDK5 silencing mediates sensitivity was investigated. Previously, CDK5 has been suggested to be active only in a neuronal context, but here we show that CDK5 is required in non-neuronal cells for the DNA-damage response and, in particular, intra-S and G(2)/M cell-cycle checkpoints. These results highlight the potential of synthetic lethal siRNA screens with chemical inhibitors to define new determinants of sensitivity and potential therapeutic targets.

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KW - Humans

KW - Hydrogen Peroxide/pharmacology

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KW - Protein-Serine-Threonine Kinases/genetics

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JO - EMBO Journal

JF - EMBO Journal

SN - 0261-4189

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ER -

A synthetic lethal siRNA screen identifying genes mediating sensitivity to a PARP inhibitor

Abstract

Keywords

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