Anti-Cancer Drug Discovery Using Synthetic Lethal Chemogenetic (SLC) Analysis

摘要

I am developing a novel cell-based small-molecule screening approach that can identify inhibitors of any non-essential protein function through a surrogate synthetic lethal phenotype in the baker's yeast, Saccharomyces cerevisiae. Synthetic lethality (SL) is a form of genetic enhancement in which two mutations are lethal in combination, but the corresponding individual mutants are viable. Thus, a sensitized yeast strain carrying a mutation that is synthetic lethal with a gene of interest will be inviable in the presence of a chemical inhibitor of the target protein. Systematic genome-wide genetic screens can simultaneously determine all the synthetic lethal genetic interactions for a given gene deletion in yeast. I will adopt this strategy to determine the SL partners for the yeast genes SCH9, the yeast homologue of the human oncogene AKT, and TEP1, the yeast equivalent of the human tumor suppressor PTEN. Selected confirmed synthetic lethal mutants will be used as sensitized strains to screen a commercial small-molecule library for inhibitors of the corresponding proteins. I have developed a yeast-based high-throughput screening platform to screen the Maybridge small-molecule library. Compounds derived from the initial chemical genetic screen will be validated biochemically and, ultimately, tested on mammalian cells for activity against the human homologues.