Use of isogenic human cancer cells for high-throughput screening and drug discovery

Christopher J. Torrance, Vijai Agrawal, Bert Vogelstein, Kenneth W Kinzler

Research output: Contribution to journalArticle

Abstract

Cell-based screening for novel tumor-specific drugs has been compromised by the lack of appropriate control cells. We describe a strategy for drug screening based on isogenic human cancer cell lines in which key tumorigenic genes have been deleted by targeted homologous recombination. As a test case, a yellow fluorescent protein (YFP) expression vector was introduced into the colon cancer cell line DLD-1, and a blue fluorescent protein (BFP) expression vector was introduced into an isogenic derivative in which the mutant K-Ras allele had been deleted. Co-culture of both cell lines allowed facile screening for compounds with selective toxicity toward the mutant Ras genotype. Among 30,000 compounds screened, a novel cytidine nucleoside analog was identified that displayed selective activity in vitro and inhibited tumor xenografts containing mutant Ras. The present data demonstrate a broadly applicable approach for mining therapeutic agents targeted to the specific genetic alterations responsible for cancer development.

Original languageEnglish (US)
Pages (from-to)940-945
Number of pages6
JournalNature Biotechnology
Volume19
Issue number10
DOIs
StatePublished - 2001

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Drug Discovery
Screening
Cells
Throughput
Tumors
Cell Line
Proteins
Cytidine
Neoplasms
Nucleosides
Cell culture
Heterografts
Pharmaceutical Preparations
Preclinical Drug Evaluations
Toxicity
Homologous Recombination
Coculture Techniques
Genes
Colonic Neoplasms
Derivatives

ASJC Scopus subject areas

  • Microbiology

Cite this

Use of isogenic human cancer cells for high-throughput screening and drug discovery. / Torrance, Christopher J.; Agrawal, Vijai; Vogelstein, Bert; Kinzler, Kenneth W.

In: Nature Biotechnology, Vol. 19, No. 10, 2001, p. 940-945.

Research output: Contribution to journalArticle

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