Ferroptosis in drug resistant glioma

耐药神经胶质瘤中的铁死亡

• 批准号: 10808297
• 负责人: Deanna Marie Tiek
• 金额: $ 12.87万
• 依托单位: NORTHWESTERN UNIVERSITY AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-19 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10808297
• 关键词: Active Sites; Advisory Committees; Affect; Animal Model; Animals; Automobile Driving; Biological; Biology; Blood - brain barrier anatomy; Brain; Brain Neoplasms; CRISPR screen; Caring; Cell Death; Cell Line; Cells; Chemicals; Clinical; Coin; Cyst; Cysteine; Cystine; DNA; DNA Damage; Data; Dependence; Disease; Disease Resistance; Drops; Drug Targeting; Drug resistance; EWSR1 gene; Electron Microscopy; Environment; Enzymes; Evaluation; Future; Glioblastoma; Glioma; Goals; Hypoxia; Image; Immunocompetent; In Vitro; Intervention; Iron; Knowledge; Learning; Lipids; Longevity; Malignant Neoplasms; Malignant neoplasm of brain; Mediating; Mentors; Metabolic; Methods; Microscopy; Mission; Mitochondria; Modeling; Monitor; Mutation; Neuroprotective Agents; Newly Diagnosed; Outcome; Oxidative Stress; Pathway interactions; Patients; Persons; Phenotype; Postdoctoral Fellow; Proteins; Proteomics; RNA; RNA Splicing; Recycling; Research; Resistance; Resources; Role; Running; Scientist; Selenium; Selenium Compounds; Structure; Sulfur; Techniques; Testing; Therapeutic Effect; Therapeutically Targetable; Training; Universities; Validation; Work; analog; cancer drug resistance; cancer type; candidate identification; cell type; chemotherapeutic agent; design; ebselen; frontier; genome sequencing; improved; in vivo; insight; lipidomics; model design; neuroprotection; novel; oxidation; patient screening; potential biomarker; response; side effect; skills; small molecule; standard of care; temozolomide; tumor xenograft; whole genome

Project Abstract Cancer drug resistance is the last frontier in extending patient overall survival, especially in the deadly brain tumor – glioblastoma (GBM). GBM currently has one standard of care chemotherapeutic agent – temozolomide (TMZ) – which extends median overall survival to ~14-16 months. However, TMZ resistance is rapid and uniformly fatal. Yet, because there are some long-term responders (~5% at 5 years), almost all newly diagnosed GBMs will receive TMZ, making TMZ-resistance a major clinical unmet need, and the focus of my previous, current, and future work. The central goal of my current work is to better understand the distinct changes in the metabolic states between TMZ-Sensitive (TMZ-S) and -Resistant (TMZ-R) disease, allowing for the discovery of therapeutically targetable vulnerabilities. To this end, I have found that TMZ-R cells have a greater dependence on cystine import, increase of intracellular iron, and a maintained oxidative state that primes these TMZ-R models for cyst(e)ine depletion-induced ferroptosis with a known neuroprotective agent – ebselen. I further show that this form of ferroptosis – or iron-dependent cell death – could be induced by the selenium in ebselen or TMZ-Selenium but not a TMZ-Sulfur analog in TMZ-R cells, which will no longer have a response to TMZ alone. Therefore, the objective of this project is to meticulously characterize this cyst(e)ine depletion- induced ferroptosis phenotype in TMZ-R models where it will both help in our understanding of TMZ-R GBM and develop a framework where small molecule selenium-containing compounds could be of use in other diseases. To accomplish this goal, I will take full advantage of my excellent mentoring/advisory committee as well as the exceptional resources and training environment available to me at Northwestern University. The expected outcomes of this project include a thorough understanding of the metabolic landscape and changes between both TMZ-S and -R models, as well as shifts post cyst(e)ine depletion-induced ferroptosis (Aim 1). A characterization and validation of the protein players that are necessary to mediate cyst(e)ine depletion-induced ferroptosis in TMZ-R models, as well as how the increased oxidative state in TMZ-R may affect enzymatic active site status that would not be discovered via sequencing methods but is essential for protein activity (Aim 2). Finally, the mechanistic knowledge gained from a meticulous understanding of both the metabolic readouts and proteomic players will allow for a smarter animal model design to answer and/or confirm the findings in Aim 1 & 2 (Aim 3). As I already have a strong background in RNA biology/splicing and DNA damage/structure in TMZ-R disease, this proposal will strengthen my knowledge in the metabolic differences and learn proteomic and metabolic techniques. In this way, I will have a well-rounded understanding of TMZ-R disease which will help me to design smarter and more complete studies in the future when I run my own independent lab.

项目摘要 癌症耐药性是延长患者总体生存期的最后前沿，尤其是在致命的情况下 脑肿瘤——胶质母细胞瘤（GBM）。 GBM 目前有一种护理标准化疗药物 – 替莫唑胺 (TMZ) – 将中位总生存期延长至约 14-16 个月。然而，TMZ 耐药性是 迅速且一致致命。然而，由于存在一些长期反应者（5 年时约 5%），几乎所有新近反应者 诊断出的 GBM 将接受 TMZ，这使得 TMZ 耐药成为主要的临床未满足需求，也是我的重点 以前、现在和将来的工作。我当前工作的中心目标是更好地理解不同的 TMZ 敏感 (TMZ-S) 和耐药 (TMZ-R) 疾病之间代谢状态的变化，从而允许 发现治疗上可靶向的脆弱性。为此，我发现 TMZ-R 细胞具有 对胱氨酸输入的更大依赖性、细胞内铁的增加以及维持的氧化状态 这些 TMZ-R 模型用于使用已知的神经保护剂依布硒啉 (ebselen) 进行半胱氨酸耗竭诱导的铁死亡。 我进一步表明，这种形式的铁死亡（或铁依赖性细胞死亡）可以由硒诱导 依布硒啉或 TMZ-硒，但不是 TMZ-R 细胞中的 TMZ-硫类似物，后者将不再对 TMZ一个人。因此，该项目的目标是仔细表征这种半胱氨酸消耗- 在 TMZ-R 模型中诱导铁死亡表型，这将有助于我们理解 TMZ-R GBM 和 开发一个框架，使小分子含硒化合物可用于其他疾病。 为了实现这一目标，我将充分利用我优秀的指导/顾问委员会以及 西北大学为我提供了卓越的资源和培训环境。预期的 该项目的成果包括对代谢格局和之间的变化的透彻了解 TMZ-S 和 -R 模型，以及半胱氨酸耗尽后诱导的铁死亡（目标 1）。一个 介导半胱氨酸耗竭所必需的蛋白质参与者的表征和验证 TMZ-R 模型中的铁死亡，以及 TMZ-R 中氧化状态的增加如何影响酶活性 位点状态无法通过测序方法发现，但对于蛋白质活性至关重要（目标 2）。 最后，通过对代谢读数和代谢读数的细致理解而获得的机械知识 蛋白质组参与者将允许更智能的动物模型设计来回答和/或确认目标 1 和中的发现 2（目标 3）。因为我已经在 TMZ-R 的 RNA 生物学/剪接和 DNA 损伤/结构方面拥有深厚的背景 疾病，这个建议将加强我对代谢差异的了解，并学习蛋白质组学和 代谢技术。这样，我将对TMZ-R疾病有一个全面的了解，这将有助于我 当我将来经营自己的独立实验室时，可以设计更智能、更完整的研究。