A cell-based screening targeting the human kinome reveals a novel synthetic lethal interaction with BRCA1.

SOFÍA CARBAJOSA; MARÍA F PANSA; NATALIA PAVIOLO; PAOLA CAMPODÓNICO; DIEGO ANDINO; IRIS A. GARCÍA; BELÉN FEDERICO; JOSÉ LUIS BOCCO; VANESA GOTTIFREDI; GASTÓN SORIA

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 thedevelopment of therapeutic approaches with selective tumor cytotoxicity.A precision strategy of this kind is the induction of syntheticlethality, which takes advantage of tumor mutations to induce cytotoxicityin tumor cells with a reduced impact on normal cells.Mutations in BRCA1 and BRCA2 genes have been establishedas drivers of breast and ovarian cancer development, and thereforeconstitute an interesting niche to search for novel synthetic lethalinteractions. Herein, we developed a high-throughput screeningplatform based on flow cytometry to identify new synthetic lethalityinducers in BRCA1 and BRCA2-deficient contexts. This platform involvesthe co-culture of BRCA1/2-deficient and proficient cells fromisogenic origin that are tagged with different fluorescent proteins.The isogenic culture is exposed to the compounds from the librariesand the relative cell survival of each cell population is assessed atthe end of the experiment. Using this platform, we screened a libraryof 684 protein kinases inhibitors and we identified a mitotic kinasethat is synthetic lethal with BRCA1. We describe here the screeningplatform, the early validation phase using different cell lines andcommercial inhibitors, and the preliminary study of the mechanismthat triggers the synthetic lethality induction in BRCA1-deficientcells.