A Cell-Based Screening Targeting the Human Kinome Reveals a Novel Synthetic Lethal Interaction with BRCA1

CARBAJOSA S.; PANSA M.F.; PAVIOLO N.S.; CAMPODÓNICO P.B; ANDINO D.; GARCÍA A.; FEDERICO B.; BOCCO J.L.; GOTTIFREDI V.; SORIA G.

Buenos Aires

Congreso; Reunión Conjunta de Sociedades de Biociencias 2017; 2017

Sociedad Argentina de Investigación en Bioquímica y Biología Molecular

One of the current challenges in the fight against cancer is the development of therapeutic approaches with selective tumor cytotoxicity.A precision strategy of this kind is the induction of synthetic lethality, which takes advantage of tumor mutations to induce cytotoxicity in tumor cells with a reduced impact on normal cells.Mutations in BRCA1 and BRCA2 genes have been established as drivers of breast and ovarian cancer development, and therefore constitute an interesting niche to search for novel synthetic lethal interactions. Herein, we developed a high-throughput screening platform based on flow cytometry to identify new synthetic lethality inducers in BRCA1 and BRCA2-deficient contexts. This platform involves the co-culture of BRCA1/2-deficient and proficient cells from isogenic origin that are tagged with different fluorescent proteins.The isogenic culture is exposed to the compounds from the libraries and the relative cell survival of each cell population is assessed at the end of the experiment. Using this platform, we screened a library of 684 protein kinases inhibitors and we identified a mitotic kinase that is synthetic lethal with BRCA1. We describe here the screening platform, the early validation phase using different cell lines and commercial inhibitors, and the preliminary study of the mechanism that triggers the synthetic lethality induction in BRCA1-deficient cells.