title: Chemogenetic profiling identifies RAD17 as synthetically lethal with checkpoint kinase inhibition.
abstract: Chemical inhibitors of the checkpoint kinases have shown promise in the treatment of cancer, yet their clinical utility may be limited by a lack of molecular biomarkers to identify specific patients most likely to respond to therapy. To this end, we screened 112 known tumor suppressor genes for synthetic lethal interactions with inhibitors of the CHEK1 and CHEK2 checkpoint kinases. We identified eight interactions, including the Replication Factor C (RFC)-related protein RAD17. Clonogenic assays in RAD17 knockdown cell lines identified a substantial shift in sensitivity to checkpoint kinase inhibition (3.5-fold) as compared to RAD17 wild-type. Additional evidence for this interaction was found in a large-scale functional shRNA screen of over 100 genotyped cancer cell lines, in which CHEK1/2 mutant cell lines were unexpectedly sensitive to RAD17 knockdown. This interaction was widely conserved, as we found that RAD17 interacts strongly with checkpoint kinases in the budding yeast Saccharomyces cerevisiae. In the setting of RAD17 knockdown, CHEK1/2 inhibition was found to be synergistic with inhibition of WEE1, another pharmacologically relevant checkpoint kinase. Accumulation of the DNA damage marker γH2AX following chemical inhibition or transient knockdown of CHEK1, CHEK2 or WEE1 was magnified by knockdown of RAD17. Taken together, our data suggest that CHEK1 or WEE1 inhibitors are likely to have greater clinical efficacy in tumors with RAD17 loss-of-function.
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.|Moores Cancer Center, University of California San Diego, La Jolla, CA, USA.
authors.forename: John Paul
authors.initials: JP
authors.lastname: Shen
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.|Department of Genetics, Stanford University School of Medicine, Stanford, CA, USA.
authors.forename: Rohith
authors.initials: R
authors.lastname: Srivas
authors.affiliation: Bioinformatics and Systems Biology Program, University of California San Diego, La Jolla, CA, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Gross
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.|University of Pittsburgh Cancer Institute, Hillman Cancer Center, Pittsburgh, PA, USA.|University of South Alabama Mitchell Cancer Institute, Mobile, AL, USA.
authors.forename: Jianfeng
authors.initials: J
authors.lastname: Li
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.|Ludwig Institute for Cancer Research, University of California San Diego, La Jolla, CA, USA.
authors.forename: Eric J
authors.initials: EJ
authors.lastname: Jaehnig
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Einthovenweg, Leiden, The Netherlands.
authors.forename: Su Ming
authors.initials: SM
authors.lastname: Sun
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Ana
authors.initials: A
authors.lastname: Bojorquez-Gomez
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Katherine
authors.initials: K
authors.lastname: Licon
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Vignesh
authors.initials: V
authors.lastname: Sivaganesh
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Jia L
authors.initials: JL
authors.lastname: Xu
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Kristin
authors.initials: K
authors.lastname: Klepper
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Huwate
authors.initials: H
authors.lastname: Yeerna
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Pekin
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.
authors.forename: Chu Ping
authors.initials: CP
authors.lastname: Qiu
authors.affiliation: Department of Toxicogenetics, Leiden University Medical Center, Einthovenweg, Leiden, The Netherlands.
authors.forename: Haico
authors.initials: H
authors.lastname: van Attikum
authors.affiliation: Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.|University of Pittsburgh Cancer Institute, Hillman Cancer Center, Pittsburgh, PA, USA.|University of South Alabama Mitchell Cancer Institute, Mobile, AL, USA.
authors.forename: Robert W
authors.initials: RW
authors.lastname: Sobol
authors.affiliation: Department of Medicine, University of California San Diego, La Jolla, CA, USA.|Moores Cancer Center, University of California San Diego, La Jolla, CA, USA.|Bioinformatics and Systems Biology Program, University of California San Diego, La Jolla, CA, USA.
authors.forename: Trey
authors.initials: T
authors.lastname: Ideker
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D054316:Biomarkers, Pharmacological / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects / Q000235:genetics; D018797:Cell Cycle Proteins / Q000235:genetics / Q000378:metabolism*; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2 / Q000235:genetics / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D055808:Drug Discovery; D006367:HeLa Cells; D006801:Humans; D058990:Molecular Targeted Therapy; D009154:Mutation / Q000235:genetics; D009369:Neoplasms / Q000175:diagnosis / Q000188:drug therapy*; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism; D011505:Protein-Tyrosine Kinases / Q000235:genetics / Q000378:metabolism; D034741:RNA, Small Interfering / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000472:pathogenicity* / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*; D013876:Thiophenes / Q000494:pharmacology*; D014508:Urea / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: C532363:3-(carbamoylamino)-5-(3-fluorophenyl)-N-(3-piperidyl)thiophene-2-carboxamide; D000970:Antineoplastic Agents; D054316:Biomarkers, Pharmacological; D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; C501395:RAD17 protein, S cerevisiae; D034741:RNA, Small Interfering; C113575:Rad17 protein, human; D029701:Saccharomyces cerevisiae Proteins; D013876:Thiophenes; D014508:Urea; D011494:Protein Kinases; C099685:SWE1 protein, S cerevisiae; D064447:Checkpoint Kinase 2; D011505:Protein-Tyrosine Kinases; C000605867:CHEK1 protein, human; D000071877:Checkpoint Kinase 1
keywords: DNA damage; RAD17; biomarker; checkpoint kinase inhibitor; synthetic lethal