Leveraging evolutionary adaptations to uncover mechanisms of oxidative stress resistance

利用进化适应揭示氧化应激抵抗机制

• 批准号: 10785198
• 负责人: JAMES A OLZMANN
• 金额: $ 41.45万
• 依托单位: UNIVERSITY OF CALIFORNIA BERKELEY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-30 至 2025-09-29
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10785198
• 关键词: Address; Aerobic; Age; Aging; Animals; Applied Genetics; Asphyxia; Biological; Biological Models; Biology; CRISPR library; CRISPR screen; CRISPR/Cas technology; Cell Culture System; Cell Death; Cells; Cellular Assay; Chemicals; Custom; Cytoprotection; Dangerousness; Data; Degenerative Disorder; Development; Disease; Disease model; Diving; Endothelial Cells; Etiology; Excision; Foundations; Future; Gene Delivery; Gene Expression; Generations; Genes; Genetic; Genetic Screening; Glutathione; Hour; Human; Hydrogen Peroxide; Injury; Interdisciplinary Study; Iron; Ischemia; Knock-out; Knowledge; Libraries; Life; Lipid Peroxidation; Lipid Peroxides; Lipids; Mammals; Measures; Mediating; Methods; Mirounga; Molecular; Molecular Biology; Nerve Degeneration; Oxidants; Oxidative Stress; Oxygen; Pathologic; Pathway interactions; Phospholipids; Physiological; Physiological Adaptation; Process; Reactive Oxygen Species; Reperfusion Injury; Research; Resistance; Resources; Role; System; Testing; Therapeutic; Tissues; Translating; age related; cell type; combat; comparative; deep sequencing; genetic analysis; genetic manipulation; genome-wide; insight; loss of function; macromolecule; novel; novel therapeutic intervention; oxidant stress; oxidative damage; pressure; prevent; reproductive; resistance mechanism; seal; stressor; therapeutic target; tissue degeneration; tissue injury; tool

PROJECT SUMMARY / ABSTRACT Imbalances in reactive oxygen species generation and removal can lead to irreversible damage to macromolecules, such as lipids. The accumulation of oxidatively damage lipids (i.e., lipid peroxides) is a hallmark feature of ferroptosis, an iron-dependent regulated form of cell death. Lipid peroxidation and ferroptosis have been implicated in the etiology of aging, age-related diseases (e.g., neurodegeneration), and other forms of tissue degeneration such as ischemia-reperfusion injury. However, the mechanisms by which cells combat lipid peroxidation and ferroptosis, as well as how these processes can be therapeutically targeted, remain poorly understood. To address this gap in knowledge, our proposed research aims to develop cultured elephant seal cells as a non-traditional comparative model system to identify new mechanisms that suppress oxidative stress and ferroptosis. Elephant seals provide an exceptional model system for questions related to oxidative stress because they have evolved a remarkable resistance to oxidative tissue damage. During dives up to 2-hours, elephant seals undergo progressive and repeated tissue ischemia, but in contrast to the ischemia-reperfusion injury that occurs in humans, elephant seal tissues are protected from damage. Furthermore, our preliminary data demonstrate that cultured elephant seal cells are resistant to a variety of oxidative stressors (e.g., ferroptosis inducers), indicating that elephant seals have evolved cell autonomous mechanisms to suppress oxidative damage and ferroptosis. Elucidating the mechanisms that provide elephant seal cells with this unique ability to prevent oxidative damage provides an extraordinary opportunity to uncover unexpected pathways that will provide insights into aging and diseases associated with oxidative stress and will reveal new potential therapeutic opportunities. Here, we propose to develop genetic editing methods and genome-wide CRISPR-Cas9 libraries to study known lipid peroxidation and ferroptosis in cultured elephant seal and human cells (Aim 1). Furthermore, we will perform genome-wide synthetic lethal screens to systematically uncover the mechanisms of ferroptosis resistance in cultured elephant seal cells (Aim 2). Completion of these studies will provide the first genome-wide CRISPR-Cas9 library for elephant seal cells and will identify novel genetic modifiers of ferroptosis, launching a new model system into the molecular era, potentially yielding insights into the etiology of degenerative diseases, and providing a foundation for the development of novel therapeutic strategies.

项目概要/摘要 活性氧产生和去除的不平衡可能会导致不可逆转的损害 大分子，例如脂质。氧化损伤脂质（即脂质过氧化物）的积累是一个标志 铁死亡的特征，铁依赖性细胞死亡的调节形式。脂质过氧化和铁死亡 与衰老、与年龄相关的疾病（例如神经退行性疾病）和其他形式的病因有关 组织退化，例如缺血再灌注损伤。然而，细胞对抗脂质的机制 过氧化和铁死亡，以及如何针对这些过程进行治疗，仍然知之甚少。 明白了。为了解决这一知识差距，我们提出的研究旨在开发养殖象海豹 细胞作为非传统比较模型系统来识别抑制氧化应激的新机制 和铁死亡。海象为氧化应激相关问题提供了一个特殊的模型系统 因为它们已经进化出对氧化组织损伤的显着抵抗力。在长达 2 小时的潜水过程中， 象海豹经历进行性和反复的组织缺血，但与缺血再灌注相反 当人类发生伤害时，象海豹组织可以免受损伤。此外，我们初步 数据表明，培养的象海豹细胞能够抵抗多种氧化应激源（例如，铁死亡） 诱导剂），表明象海豹已经进化出细胞自主机制来抑制氧化 损伤和铁死亡。阐明象海豹细胞具有这种独特能力的机制 防止氧化损伤提供了一个绝佳的机会来发现意想不到的途径， 提供对衰老和与氧化应激相关的疾病的见解，并将揭示新的潜在治疗方法 机会。在这里，我们建议开发基因编辑方法和全基因组CRISPR-Cas9文库 研究培养象海豹和人类细胞中已知的脂质过氧化和铁死亡（目标 1）。此外， 我们将进行全基因组合成致死筛选，系统地揭示铁死亡的机制 培养的象海豹细胞的耐药性（目标 2）。这些研究的完成将提供第一个全基因组 用于象海豹细胞的 CRISPR-Cas9 文库将鉴定铁死亡的新型基因修饰剂，推出 进入分子时代的新模型系统，有可能深入了解退行性疾病的病因学， 并为开发新的治疗策略提供基础。