title: Parallel in vivo and in vitro melanoma RNAi dropout screens reveal synthetic lethality between hypoxia and DNA damage response inhibition.
abstract: To identify factors preferentially necessary for driving tumor expansion, we performed parallel in vitro and in vivo negative-selection short hairpin RNA (shRNA) screens. Melanoma cells harboring shRNAs targeting several DNA damage response (DDR) kinases had a greater selective disadvantage in vivo than in vitro, indicating an essential contribution of these factors during tumor expansion. In growing tumors, DDR kinases were activated following hypoxia. Correspondingly, depletion or pharmacologic inhibition of DDR kinases was toxic to melanoma cells, including those that were resistant to BRAF inhibitor, and this could be enhanced by angiogenesis blockade. These results reveal that hypoxia sensitizes melanomas to targeted inhibition of the DDR and illustrate the utility of in vivo shRNA dropout screens for the identification of pharmacologically tractable targets.
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Patricia A
authors.initials: PA
authors.lastname: Possik
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Judith
authors.initials: J
authors.lastname: Müller
authors.affiliation: Division of Immunology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Carmen
authors.initials: C
authors.lastname: Gerlach
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Juliana C N
authors.initials: JC
authors.lastname: Kenski
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Xinyao
authors.initials: X
authors.lastname: Huang
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Aida
authors.initials: A
authors.lastname: Shahrabi
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Oscar
authors.initials: O
authors.lastname: Krijgsman
authors.affiliation: Division of Experimental Animal Pathology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Ji-Ying
authors.initials: JY
authors.lastname: Song
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Marjon A
authors.initials: MA
authors.lastname: Smit
authors.affiliation: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Gerritsen
authors.affiliation: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Cor
authors.initials: C
authors.lastname: Lieftink
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Kristel
authors.initials: K
authors.lastname: Kemper
authors.affiliation: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Magali
authors.initials: M
authors.lastname: Michaut
authors.affiliation: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Roderick L
authors.initials: RL
authors.lastname: Beijersbergen
authors.affiliation: Division of Molecular Carcinogenesis, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Lodewyk
authors.initials: L
authors.lastname: Wessels
authors.affiliation: Division of Immunology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands.
authors.forename: Ton N
authors.initials: TN
authors.lastname: Schumacher
authors.affiliation: Division of Molecular Oncology, The Netherlands Cancer Institute, 1066 CX Amsterdam, the Netherlands. Electronic address: d.peeper@nki.nl.
authors.forename: Daniel S
authors.initials: DS
authors.lastname: Peeper
mesh_terms: D000818:Animals; D015687:Cell Hypoxia / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2 / Q000378:metabolism; D004249:DNA Damage; D004260:DNA Repair / Q000187:drug effects; D005820:Genetic Testing; D057809:HEK293 Cells; D006801:Humans; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000378:metabolism; D008545:Melanoma / Q000235:genetics* / Q000473:pathology*; D051379:Mice; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D011494:Protein Kinases / Q000378:metabolism; D055550:Protein Stability / Q000187:drug effects; D034622:RNA Interference / Q000187:drug effects; D034741:RNA, Small Interfering / Q000378:metabolism; D015203:Reproducibility of Results; D015398:Signal Transduction / Q000187:drug effects; D014407:Tumor Cells, Cultured; D023041:Xenograft Model Antitumor Assays
chemical_list: D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D047428:Protein Kinase Inhibitors; D034741:RNA, Small Interfering; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; D000071877:Checkpoint Kinase 1