Identification Of Novel Essential Genes For Prostate Cancer Metastasis By Genome Scale Crispr Approaches.

Abstract Background: Thanks to the availability of new biomarkers, Prostate cancer (PCa) can now be diagnosed earlier and, as a result, PCa patients have a high 5-year survival rate. Nonetheless, the situation changes completely for patients who develop metastasis, for whom 5-year survival rate decreases to 29%. Recent candidate-gene approaches have identified new candidates, but there are still no drugs to prevent or cure metastatic PCa. Hence, there is an urgent need to find effective therapeutic targets. Methodology: In this study we have conducted a high-throughput CRISPR/Cas9 screening in highly metastatic PCa cell lines PC3 and DU145, using the CRISPR/Cas9 knock-out (GeCKO) library, that targets 18,080 genes with 64,751 unique gRNAs. To assess whether each gene loss-of-function may be critical for metastasis development, invasion assays in Matrigel-coated Boyden Chamber membranes were used. Next-generation sequencing and bioinformatic MaGECK analysis were then carried out to identify novel key genes in the development of PCa metastasis. siRNA approach was used to further validate the best candidates. Results: We found 29 candidates and several signaling pathways capable of significatively impair the invasion of, both, PC3 and DU145 cell lines when knocked-out, being PRMT7 the strongest regulator. PRMT7 gene encodes a type II methyltransferase involved in several cellular processes such as mRNA splicing or DNA repair. There are evidences that indicate it regulates EMT through methylation of E-cadherin promoter and other targets. Moreover, its overexpression correlates with higher levels of expression of MMP9 and MMP2 that lead to metastasis appearance in other types of tumors. Our results inhibiting PRMT7 expression with siRNA validated our high-throughput CRISPR/Cas9 screening results and confirmed the implication of PRMT7 in PCa invasion. Conclusions: We have conducted a novel high-throughput CRISPR-Cas9 screening, finding promising gene candidates, such as PRMT7, which, in the future could be used as therapeutical targets to prevent metastasis development in PCa patients. Furthermore, in this study we have also found genes that had been previously associated to PCa metastasis in other candidate-gene studies, giving a proof-of-concept for the use of high performance of CRISPR-Cas9 screenings for the search of candidate genes involved in PCa metastasis. Citation Format: Maria Rodrigo Faus, Sara Manzano Figueroa, Jingchun Qu, Huiqi Qu, Renata Pellegrino, Hakon Hakonarson, Paloma Bragado, Almudena Porras, Alvaro Gutierrez-Uzquiza. Identification of novel essential genes for prostate cancer metastasis by genome scale CRISPR approaches [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2310.