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

项目概要和摘要 这个NRSA个人奖学金的广泛目标是双重的：1）促进发展， 基本技能，使候选人成为一个有效的医生，科学家，和2）调查 基于CRISPR的筛选揭示了癌症氧化还原稳态的机制。候选人及其 导师们设计了一个具体的培训计划，以实现这些目标。该计划包括严格的 研究的组成部分，奠定了成功的职业生涯的基础。这件事跟龙先生有关 腺癌和氧化应激的调节。肺癌仍然是癌症死亡的主要原因 在男性和女性中。最近成功的代谢药物，如复合物I抑制剂， 重新激发了人们对靶向癌症代谢的兴趣。然而，这些药物中有许多都有令人难以接受的一面 方面的影响.更深入地了解癌症代谢重编程对于癌症的发展是必要的。 更有效更安全的代谢靶向癌症疗法。 最近的研究发现，十字花科蔬菜的饮食消费，如西兰花， 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酯 例如异硫氰酸苯乙酯（PEITC）。尽管已知PEITC破坏了微妙的癌症氧化还原过程， 平衡，其发生的机制是未知的。候选人有初步的数据利用一个 基于代谢聚焦的CRISPR的筛选揭示了PEITC诱导的肺疾病所需的KDM5C 腺癌细胞死亡和SCD 1作为与PEITC的合成致死。本提案的重点是验证 这些基因命中并检查KDM5C和SCD 1调节细胞抗氧化剂的机制， 反应这项提案的长期目标是确定治疗代谢目标，可能导致更多的 肺癌的治疗方法有哪些？ 在具体目标1中，候选人将确定PEITC诱导的肺 通过抑制抗氧化基因导致腺癌细胞死亡。KDM5C对组蛋白调控的分析 修饰及其对基因表达的下游影响将用于确定 KDM5C是细胞抗氧化反应的负调节剂。在具体目标2中，候选人将确定 SCD1的抑制是与PEITC合成致死以诱导肺的分子机制 腺癌细胞死亡。脂质组学将量化SCD 1调节细胞脂质水平的程度。 ER和线粒体应激的测量将提供对SCD 1被激活的机制的深入了解。 用PEITC合成致死。拟议的研究将确定和因果验证以前未知的 癌症细胞氧化还原稳态机制并为设计有效的组合疗法提供证据 针对这种平衡的疗法。

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.