High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma

Nicholas C. Wolff, Andrea Pavía-Jiménez, Vanina T. Tcheuyap, Shane Alexander, Mridula Vishwanath, Alana Christie, Xian Jin Xie, Noelle S. Williams, Payal Kapur, Bruce Posner, Renée M. McKay, James Brugarolas

Research output: Contribution to journalArticle

11 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)16951-16962
Number of pages12
JournalOncotarget
Volume6
Issue number19
StatePublished - 2015

Fingerprint

Renal Cell Carcinoma
Neoplasms
Lethal Genes
Synthetic Genes
Kidney Neoplasms
Genetic Models
Leukemia, Myelogenous, Chronic, BCR-ABL Positive
Tumor Cell Line
Fluorescent Dyes
Pharmaceutical Preparations
Apoptosis
homoharringtonine
Growth
Genes
Therapeutics

Keywords

  • High-content drug screen
  • Omacetaxine mepesuccinate
  • Patiend-derived xenografts
  • Tumorgrafts

ASJC Scopus subject areas

  • Oncology

Cite this

Wolff, N. C., Pavía-Jiménez, A., Tcheuyap, V. T., Alexander, S., Vishwanath, M., Christie, A., ... Brugarolas, J. (2015). High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma. Oncotarget, 6(19), 16951-16962.

High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma. / Wolff, Nicholas C.; Pavía-Jiménez, Andrea; Tcheuyap, Vanina T.; Alexander, Shane; Vishwanath, Mridula; Christie, Alana; Xie, Xian Jin; Williams, Noelle S.; Kapur, Payal; Posner, Bruce; McKay, Renée M.; Brugarolas, James.

In: Oncotarget, Vol. 6, No. 19, 2015, p. 16951-16962.

Research output: Contribution to journalArticle

Wolff NC, Pavía-Jiménez A, Tcheuyap VT, Alexander S, Vishwanath M, Christie A et al. High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma. Oncotarget. 2015;6(19):16951-16962.
@article{9e67eb437a60415fad7ec408ca80b819,
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.",
keywords = "High-content drug screen, Omacetaxine mepesuccinate, Patiend-derived xenografts, Tumorgrafts",
author = "Wolff, {Nicholas C.} and Andrea Pav{\'i}a-Jim{\'e}nez and Tcheuyap, {Vanina T.} and Shane Alexander and Mridula Vishwanath and Alana Christie and Xie, {Xian Jin} and Williams, {Noelle S.} and Payal Kapur and Bruce Posner and McKay, {Ren{\'e}e M.} and James Brugarolas",
year = "2015",
language = "English (US)",
volume = "6",
pages = "16951--16962",
journal = "Oncotarget",
issn = "1949-2553",
publisher = "Impact Journals",
number = "19",

}

TY - JOUR

T1 - High-throughput simultaneous screen and counterscreen identifies homoharringtonine as synthetic lethal with von Hippel-Lindau loss in renal cell carcinoma

AU - Wolff, Nicholas C.

AU - Pavía-Jiménez, Andrea

AU - Tcheuyap, Vanina T.

AU - Alexander, Shane

AU - Vishwanath, Mridula

AU - Christie, Alana

AU - Xie, Xian Jin

AU - Williams, Noelle S.

AU - Kapur, Payal

AU - Posner, Bruce

AU - McKay, Renée M.

AU - Brugarolas, James

PY - 2015

Y1 - 2015

N2 - 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.

AB - 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.

KW - High-content drug screen

KW - Omacetaxine mepesuccinate

KW - Patiend-derived xenografts

KW - Tumorgrafts

UR - http://www.scopus.com/inward/record.url?scp=84938248489&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84938248489&partnerID=8YFLogxK

M3 - Article

VL - 6

SP - 16951

EP - 16962

JO - Oncotarget

JF - Oncotarget

SN - 1949-2553

IS - 19

ER -