Investigating Mechanisms of Cancer Redox Homeostasis with Metabolic CRISPR-Based Screens

通过基于代谢 CRISPR 的筛选研究癌症氧化还原稳态机制

• 批准号: 10216160
• 负责人: Karthik Vasan
• 金额: $ 5.1万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10216160
• 关键词: A549; Adenocarcinoma Cell; Antioxidants; Binding Sites; Biology; Broccoli - dietary; CRISPR screen; Cancer Etiology; Cancer cell line; Cell Death; Cell Respiration; Cells; Cessation of life; ChIP-seq; Chemoprotective Agent; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Complex; Consumption; Data; Development; Environment; Equilibrium; Fatty Acids; Fellowship; Foundations; Gene Expression; Genes; Genetic Screening; Glucose; Goals; Guide RNA; Homeostasis; Human; Hypoxia; In Vitro; Individual; Isothiocyanates; Lead; Lipids; Lung; Lung Adenocarcinoma; Malignant Neoplasms; Malignant neoplasm of lung; Measurement; Mentors; Metabolic; Metabolic stress; Metabolism; Mitochondria; Molecular; Monounsaturated Fatty Acids; Mus; Mutation; National Research Service Awards; Nutrient; Oxidants; Oxidation-Reduction; Oxidative Regulation; Oxidative Stress; Paraquat; Patient-Focused Outcomes; Patients; Pharmaceutical Preparations; Phase II Clinical Trials; Phenethyl Isothiocyanate; Physicians; Production; Property; RNA library; Radiation; Reactive Oxygen Species; Regulation; Research; Risk; Role; Saturated Fatty Acids; Scientist; Signal Pathway; Stress; Testing; Therapeutic; Training; Woman; base; biological adaptation to stress; cancer cell; cancer risk; career; cell behavior; cell transformation; combinatorial; cruciferous vegetable; demethylation; design; dietary; effective therapy; genetic selection; histone modification; improved; in vivo; inhibitor/antagonist; insight; interest; lipidomics; lung cancer cell; men; neoplastic cell; novel; overexpression; oxidative damage; repaired; response; screening; side effect; skills; small molecule inhibitor; success; targeted cancer therapy; therapeutic target; transcription factor; translational cancer research; tumor; tumor metabolism

Project Summary and Abstract The broad objectives of this NRSA Individual Fellowship are two-fold: 1) to facilitate the development of essential skills that will allow the candidate to become an effective physician-scientist, and 2) to investigate mechanisms of cancer redox homeostasis uncovered by CRISPR-based screens. The candidate and his mentor have designed a specific training plan to achieve these objectives. The plan includes a rigorous research component that lays the foundation for a successful career. This proposal concerns lung adenocarcinoma and regulation of oxidative stress. Lung cancer remains the leading cause of cancer death among both men and women. The recent success of metabolic drugs, such as complex I inhibitors, have revitalized interest in targeting cancer metabolism. However, many of these drugs have unacceptable side effects. A more thorough understanding of cancer metabolic reprogramming is necessary for the development of effective and safer, metabolically targeted cancer therapies. Recent studies have uncovered that the dietary consumption of cruciferous vegetables, such as broccoli, is associated with a decreased risk of cancer. Their chemoprotective properties are attributed to isothiocyanates such as phenethyl isothiocyanate (PEITC). Although it known that PEITC disrupts the delicate cancer redox balance, the mechanism by which this occurs is unknown. The candidate has preliminary data utilizing a metabolism focused CRISPR-based screen revealing KDM5C as required for PEITC-induced lung adenocarcinoma cell death and SCD1 as synthetic lethal with PEITC. The focus of this proposal is to validate these gene hits and examine the mechanisms by which KDM5C and SCD1 regulate the cellular antioxidant response. The long-term goal of this proposal is to identify therapeutic metabolic targets that may lead to more effective and safer therapies for patients with lung cancer. In Specific Aim 1, the candidate will determine whether KDM5C is required for PEITC-induced lung adenocarcinoma cell death by repressing antioxidant genes. Analysis of KDM5C regulation of histone modifications and their downstream effects on gene expression will be used to determine the extent to which KDM5C is a negative regulator of cellular antioxidant response. In Specific Aim 2, the candidate will determine the molecular mechanism by which inhibition of SCD1 is synthetic lethal with PEITC to induce lung adenocarcinoma cell death. Lipidomics will quantify the extent to which SCD1 regulates cellular lipid levels. Measurements of ER and mitochondrial stress will provide insight into the mechanism by which SCD1 is synthetic lethal with PEITC. The proposed studies will identify and causally validate previously unknown mechanisms of cellular redox homeostasis in cancer and provide evidence to devise effective combinatorial therapies to target this balance.

项目概要和摘要 该 NRSA 个人奖学金的广泛目标有两个：1）促进 使候选人成为一名有效的医师科学家的基本技能，以及 2) 进行调查 基于 CRISPR 的筛选揭示了癌症氧化还原稳态机制。候选人和他的 导师设计了具体的培训计划来实现这些目标。该计划包括严格的 研究部分为成功的职业生涯奠定了基础。该提案涉及肺 腺癌和氧化应激的调节。肺癌仍然是癌症死亡的主要原因 男性和女性之间。代谢药物（例如复合物 I 抑制剂）最近取得的成功 重新激发了人们对针对癌症代谢的兴趣。然而，许多这些药物都有不可接受的副作用 影响。更深入地了解癌症代谢重编程对于开发是必要的 有效且更安全的代谢靶向癌症疗法。 最近的研究发现，十字花科蔬菜（例如西兰花）的饮食摄入量 与癌症风险降低有关。它们的化学保护特性归因于异硫氰酸盐 例如异硫氰酸苯乙酯（PEITC）。尽管众所周知 PEITC 会破坏微妙的癌症氧化还原 平衡，但发生这种情况的机制尚不清楚。候选人利用以下方法获得了初步数据 基于代谢的 CRISPR 筛选揭示了 PEITC 诱导肺所需的 KDM5C 腺癌细胞死亡和 SCD1 作为 PEITC 的合成致死细胞。该提案的重点是验证 这些基因命中并检查 KDM5C 和 SCD1 调节细胞抗氧化剂的机制 回复。该提案的长期目标是确定可能导致更多的治疗代谢目标 为肺癌患者提供有效且安全的治疗方法。 在具体目标 1 中，考生将确定 PEITC 诱导的肺病是否需要 KDM5C 通过抑制抗氧化基因导致腺癌细胞死亡。 KDM5C对组蛋白的调控分析 修饰及其对基因表达的下游影响将用于确定修饰的程度 KDM5C 是细胞抗氧化反应的负调节因子。在具体目标 2 中，候选人将确定 抑制SCD1与PEITC合成致死诱导肺损伤的分子机制 腺癌细胞死亡。脂质组学将量化 SCD1 调节细胞脂质水平的程度。 ER 和线粒体应激的测量将有助于深入了解 SCD1 的机制 PEITC 合成致死剂。拟议的研究将识别并因果验证以前未知的 癌症中细胞氧化还原稳态的机制，并为设计有效的组合提供证据 针对这种平衡的疗法。