CRISPR/CAS9 Methodologies as a tool for investigating Fanconi Anemia pathway in cancer: perspective on pancreatic cancer treatment

The concept of precision medicine is advancing the discovery of new therapeutic targets for cancer treatment, offering tailored approaches based on the genetic profile of individual patients. One such approach is the paradigm of Synthetic Lethality, where cancer cells with specific genetic mutations are targeted by therapies that spare normal, healthy cells. This concept has been successfully applied in the treatment of various cancers, including ovarian, breast, and pancreatic cancers. The first synthetic lethality-based treatment to gain approval was for patients with BRCA1/2 mutations, using PARP inhibitors. These drugs disrupt the DNA repair machinery of cancer cells by inhibiting single-strand break repair, while the BRCA1/2 mutations prevent double-strand break repair. However, a major limitation of this approach is the development of resistance to PARP inhibitors, and the fact that not all cancer patients carry BRCA1/2 mutations. Therefore, there is a need to identify new synthetic lethal partners to expand the therapeutic potential of PARP inhibitors. The Fanconi Anemia pathway is involved in DNA repair and has a well-established connection with BRCA2-mediated repair processes. This PhD thesis aims to explore and validate advanced genomic methodologies to investigate the genes involved in the FA pathway, with a particular emphasis on their relationship with PARP inhibitor treatment. The ultimate goal is to identify therapeutic targets that could improve patient stratification for personalized cancer therapies, with a specific focus on pancreatic cancer cells. This study primarily focuses on utilizing genomic tools, specifically CRISPRSelect and CRISPR interference (CRISPRi), to investigate the Fanconi Anemia pathway and its relationship with sensitivity to the PARP inhibitor Talazoparib. The key genes explored include FANCA, FANCC, FANCD2, and FANCM.