CRISPR-mediated modeling and functional validation of candidate tumor suppressor genes in small cell lung cancer

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 117; no. 1; pp. 513 - 521
• Main Authors: Ng, Sheng Rong, Rideout, William M., Akama-Garren, Elliot H., Bhutkar, Arjun, Mercer, Kim L., Schenkel, Jason M., Bronson, Roderick T., Jacks, Tyler
• Format: Journal Article
• Language: English
• Published: United States National Academy of Sciences 07.01.2020
• Subjects: Animal models; Animals; Apoptosis; Apoptosis - genetics; Biological Sciences; Cell Line; Cell Proliferation - genetics; CRISPR; CRISPR-Cas Systems - genetics; Disease Models, Animal; Disease Progression; Feasibility Studies; Gene Editing - methods; Gene Expression Regulation, Neoplastic; Gene sequencing; Genes; Genes, Tumor Suppressor; Humans; Loss of Function Mutation; Lung - pathology; Lung cancer; Lung Neoplasms - genetics; Lung Neoplasms - pathology; Metastases; Mice; Mice, Transgenic; Mutation; Neoplasm Staging; Retinoblastoma-Like Protein p107 - genetics; Retinoblastoma-Like Protein p130 - genetics; Small cell lung carcinoma; Small Cell Lung Carcinoma - genetics; Small Cell Lung Carcinoma - pathology; Solid tumors; Tumor Burden - genetics; Tumor suppressor genes; Tumor Suppressor Protein p53 - genetics; Tumors; CRISPR; p107; small cell lung cancer; GEMM
• ISSN: 0027-8424; 1091-6490; 1091-6490
• DOI: 10.1073/pnas.1821893117

Small cell lung cancer (SCLC) is a highly aggressive subtype of lung cancer that remains among the most lethal of solid tumor malignancies. Recent genomic sequencing studies have identified many recurrently mutated genes in human SCLC tumors. However, the functional roles of most of these genes remain to be validated. Here, we have adapted the CRISPR-Cas9 system to a well-established murine model of SCLC to rapidly model loss-of-function mutations in candidate genes identified from SCLC sequencing studies. We show that loss of the gene p107 significantly accelerates tumor progression. Notably, compared with loss of the closely related gene p130, loss of p107 results in fewer but larger tumors as well as earlier metastatic spread. In addition, we observe differences in proliferation and apoptosis as well as altered distribution of initiated tumors in the lung, resulting from loss of p107 or p130. Collectively, these data demonstrate the feasibility of using the CRISPR-Cas9 system to model loss of candidate tumor suppressor genes in SCLC, and we anticipate that this approach will facilitate efforts to investigate mechanisms driving tumor progression in this deadly disease.