title: High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma.
abstract: Renal cell carcinoma (RCC) accounts for 85% of primary renal neoplasms, and is rarely curable when metastatic. Approximately 70% of RCCs are clear-cell type (ccRCC), and in >80% the von Hippel-Lindau (VHL) gene is mutated or silenced. We developed a novel, high-content, screening strategy for the identification of small molecules that are synthetic lethal with genes mutated in cancer. In this strategy, the screen and counterscreen are conducted simultaneously by differentially labeling mutant and reconstituted isogenic tumor cell line pairs with different fluorochromes and using a highly sensitive high-throughput imaging-based platform. This approach minimizes confounding factors from sequential screening, and more accurately replicates the in vivo cancer setting where cancer cells are adjacent to normal cells. A screen of ~12,800 small molecules identified homoharringtonine (HHT), an FDA-approved drug for treating chronic myeloid leukemia, as a VHL-synthetic lethal agent in ccRCC. HHT induced apoptosis in VHL-mutant, but not VHL-reconstituted, ccRCC cells, and inhibited tumor growth in 30% of VHL-mutant patient-derived ccRCC tumorgraft lines tested. Building on a novel screening strategy and utilizing a validated RCC tumorgraft model recapitulating the genetics and drug responsiveness of human RCC, these studies identify HHT as a potential therapeutic agent for a subset of VHL-deficient ccRCCs.
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine - Hematology-Oncology Division, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Nicholas C
authors.initials: NC
authors.lastname: Wolff
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine - Hematology-Oncology Division, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Andrea
authors.initials: A
authors.lastname: Pavía-Jiménez
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine - Hematology-Oncology Division, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Vanina T
authors.initials: VT
authors.lastname: Tcheuyap
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine - Hematology-Oncology Division, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Shane
authors.initials: S
authors.lastname: Alexander
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|BioTek Instruments, Winooski, VT, USA.
authors.forename: Mridula
authors.initials: M
authors.lastname: Vishwanath
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Alana
authors.initials: A
authors.lastname: Christie
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Xian-Jin
authors.initials: XJ
authors.lastname: Xie
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Noelle S
authors.initials: NS
authors.lastname: Williams
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Pathology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Payal
authors.initials: P
authors.lastname: Kapur
authors.affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Bruce
authors.initials: B
authors.lastname: Posner
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: Renée M
authors.initials: RM
authors.lastname: McKay
authors.affiliation: Kidney Cancer Program, Simmons Comprehensive Cancer Center, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Internal Medicine - Hematology-Oncology Division, University of Texas Southwestern Medical Center, Dallas, Texas, USA.|Department of Developmental Biology, University of Texas Southwestern Medical Center, Dallas, Texas, USA.
authors.forename: James
authors.initials: J
authors.lastname: Brugarolas
mesh_terms: D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D002292:Carcinoma, Renal Cell / Q000235:genetics*; D045744:Cell Line, Tumor; D004354:Drug Screening Assays, Antitumor / Q000379:methods; D005260:Female; D006248:Harringtonines / Q000494:pharmacology*; D057166:High-Throughput Screening Assays / Q000379:methods; D000077863:Homoharringtonine; D006801:Humans; D007680:Kidney Neoplasms / Q000235:genetics*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D051794:Von Hippel-Lindau Tumor Suppressor Protein / Q000235:genetics*; D023041:Xenograft Model Antitumor Assays
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D006248:Harringtonines; D000077863:Homoharringtonine; D051794:Von Hippel-Lindau Tumor Suppressor Protein; C497446:VHL protein, human
keywords: high-content drug screen; omacetaxine mepesuccinate; patiend-derived xenografts; tumorgrafts