Genome-wide synthetic lethal screening for vulnerabilities in a cell model of succinate dehydrogenase-loss paraganglioma

全基因组合成致死筛查琥珀酸脱氢酶缺失副神经节瘤细胞模型中的漏洞

• 批准号: 10572019
• 负责人: LOUIS JAMES MAHER
• 金额: $ 22.3万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-08 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10572019
• 关键词: Address; Allografting; Alzheimer' s Disease; Automobile Driving; Bar Codes; Breast Cancer Cell; CRISPR interference; CRISPR screen; Candidate Disease Gene; Cell Line; Cell model; Cells; Chromaffin Cells; Citric Acid Cycle; Clustered Regularly Interspaced Short Palindromic Repeats; DNA; Data Analyses; Defect; Diabetes Mellitus; Dioxygenases; Disease; Electron Transport; Enzymes; Estrogen Receptor alpha; Estrogen Receptor beta; Family; Gastrointestinal Stromal Tumors; Gene Expression; Gene Targeting; Genes; Genomic DNA; Growth; Guide RNA; Heart Diseases; Hereditary Paraganglioma; Histones; Hypoxia; Inherited; Lethal Genes; Link; Malignant Neoplasms; Measures; Metabolic; Methods; Mitochondria; Modeling; Molecular; Monitor; Mus; Mutation; Paraganglioma; Pathologic; Pathway interactions; Patients; Pheochromocytoma; Positioning Attribute; Pre-Clinical Model; RNA library; Recipe; Renal carcinoma; Resistance; Running; Signal Transduction; Succinate Dehydrogenase; Succinates; Synthetic Genes; Testing; Therapeutic; Time; Tissues; Transcription Repressor; Tumor Suppressor Genes; Validation; Work; candidate identification; cell growth; cell type; clinical application; deep sequencing; demethylation; experimental study; gene repression; genome-wide; hormone therapy; insight; knock-down; molecular pathology; neoplastic cell; neuroendocrine cancer; novel; novel therapeutics; promoter; rare cancer; screening; targeted treatment; tumor; tumorigenesis

Abstract: Genome-wide synthetic lethal screening for vulnerabilities in a cell model of succinate dehydrogenase-loss paraganglioma This proposal focuses on the molecular pathology of familial paraganglioma (PGL). Paradoxically, this remarkable neuroendocrine cancer is caused by inherited mutations that inactivate succinate dehydrogenase (SDH), an enzyme of the mitochondrial tricarboxylic acid (TCA) cycle. A fundamental scientific understanding of the molecular basis of this tumor is lacking, limiting powerful approaches that might be identified to exploit unique vulnerabilities due to the fundamental metabolic defect in this cancer. The links between SDH loss and tissue-specific tumorigenesis remain unknown, and conventional preclinical models have been unavailable. The current hypothesis for PGL tumorigenesis invokes loss or inactivation of both parental copies of any of the four SDH subunit genes (A-D). The subsequent accumulation of succinate competitively inhibits the activities of several dioxygenase enzymes that normally suppress hypoxic signaling and demethylate histones and DNA. Despite this general mechanistic model, other mechanisms are possible and it is unclear what unique vulnerabilities may be present in SDH-loss cells that could permit targeted therapies. We hypothesize that an unbiased genome-wide lentiviral CRISPR screen will identify genes whose loss displays synthetic lethality with SDH loss. This hypothesis is supported by previous successful work in our labs identifying genes whose loss confers resistance to multiple forms of endocrine therapy in ERα+ and ERβ+ breast cancer cells. Important new PGL cell models have become available in the form of Sdhblox/lox and Sdhb-/- immortalized mouse chromaffin cells (imCC). This positions us ideally to conduct and analyze an unbiased genome-wide synthetic lethal screen using an available CRISPR single guide RNA (sgRNA) library. Aim 1 will undertake a paired unbiased lentiviral CRISPR screen in matched normal and SDH-loss imCC lines. Aim 2 will complete data analysis to identify candidate synthetic lethal genes. Aim 3 will complete independent validation of representative synthetic lethal genes and pathways. Finally, Aim 4 will implement a mouse allograft tumor model to monitor selective inhibitory effects of nominated target gene knockdown in Sdhb-/- vs. Sdhblox/lox imCC. This project is unique in being the first unbiased synthetic lethal screen addressing the unmet cancer needs of SDH-loss familial PGL patients. Because SDH genes are tumor suppressors in other cancers, including gastrointestinal stromal tumor (GIST) and some kidney cancers, identifying potential vulnerabilities in SDH-loss cells may be broadly applicable clinically.

琥珀酸盐细胞模型的全基因组合成致死筛选