Disruption of RNA Splicing Increases Vulnerability of Cells to DNA-PK Inhibitors

• Published in: International journal of molecular sciences 2024-11, Vol.25 (21), p.11810
• Main Authors: Kovina, Anastasia P, Luzhin, Artem V, Tatarskiy, Victor V, Deriglazov, Dmitry A, Petrova, Natalia V, Petrova, Nadezhda V, Kondratyeva, Liya G, Kantidze, Omar L, Razin, Sergey V, Velichko, Artem K
• Format: Article
• Language: English
• Subjects: Cancer; Cancer therapies; Care and treatment; Cell cycle; Cell growth; Cell Line, Tumor; Cell Survival - drug effects; Cell Survival - genetics; Chemotherapy; CRISPR; CRISPR-Cas Systems; Cyclin-dependent kinases; DNA; DNA damage; DNA replication; DNA Replication - drug effects; DNA-Activated Protein Kinase - antagonists & inhibitors; DNA-Activated Protein Kinase - genetics; DNA-Activated Protein Kinase - metabolism; Genes; Genetic aspects; Genomes; Health aspects; Humans; Kinases; Medical prognosis; Medical research; Medicine, Experimental; Messenger RNA; Metabolism; Metastasis; Neoplasms - drug therapy; Neoplasms - genetics; Neoplasms - metabolism; Neoplasms - pathology; Protein Kinase Inhibitors - pharmacology; Protein kinases; Proteins; Regulation; RNA Splicing - drug effects; Scientific equipment and supplies industry
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms252111810

DNA-dependent protein kinase (DNA-PK) is a key effector of non-homologous end joining (NHEJ)-mediated double-strand break (DSB) repair. Since its identification, a substantial body of evidence has demonstrated that DNA-PK is frequently overexpressed in cancer, plays a critical role in tumor development and progression, and is associated with poor prognosis in cancer patients. Recent studies have also uncovered novel functions of DNA-PK, shifting the paradigm of the role of DNA-PK in oncogenesis and renewing interest in targeting DNA-PK for cancer therapy. To gain genetic insight into the cellular pathways requiring DNA-PK activity, we used a CRISPR/Cas9 screen to identify genes in which defects cause hypersensitivity to DNA-PK inhibitors. We identified over one hundred genes involved in DNA replication, cell cycle regulation, and RNA processing that promoted cell survival when DNA-PK kinase activity was suppressed. This gene set will be useful for characterizing novel biological processes that require DNA-PK activity and identifying predictive biomarkers of response to DNA-PK inhibition in the clinic. We also validated several genes from this set and reported previously undescribed genes that modulate the response to DNA-PK inhibitors. In particular, we found that compromising the mRNA splicing pathway led to marked hypersensitivity to DNA-PK inhibition, providing a possible rationale for the combined use of splicing inhibitors and DNA-PK inhibitors for cancer therapy.