Use of Isogenic Human Cancer Cells for High-throughput Screening and Drug Discovery
A novel high-throughput drug screening strategy is described using isogenic human cancer cell lines that are genetically identical except for the presence or absence of a key cancer causing gene. Isogenic cell lines are ?tagged? with two different colored fluorescent proteins so that the growth and survival of each cell type in a co-culture format can be monitored independently and in real time. In this way, thousands of test compounds/week can be screened for selective activity towards human cells containing a tumorigenic vs normal genotype. In a test case, a blue fluorescent protein was stably introduced into the colon cancer cell line DLD-1, which harbors the cancer causing gene mutant K-Ras, and a yellow fluorescent protein expression vector was introduced into an isogenic derivative of this line in which the mutant c-Ki-Ras oncogene had been deleted. Co-culture of both cell lines in a high-density plating format allowed facile screening for compounds with selective toxicity towards cancer cells containing the mutant Ras genotype. Among 30,000 compounds screened, two novel compounds were identified. In addition to the potential therapeutic value of these lead compounds, the present data demonstrate a broadly applicable approach for mining therapeutic agents targeted to the specific genetic alterations responsible for cancer developmentInventor(s): Torrance, Chris
Type of Offer: Licensing
« More Medical Patents