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.

项目总结/摘要 活性氧物质产生和去除的不平衡可导致不可逆的损伤， 大分子，如脂质。氧化损伤脂质的积累（即，脂质过氧化物）是一个标志 铁凋亡的特征，铁依赖性调节形式的细胞死亡。脂质过氧化和铁缺乏 与衰老的病因学有关，与年龄有关的疾病（例如，神经变性），以及其他形式的 组织变性如缺血-再灌注损伤。然而，细胞对抗脂质的机制 过氧化和铁凋亡，以及如何治疗这些过程，仍然很差， 明白为了解决这一知识差距，我们提出的研究旨在开发养殖的象海豹 细胞作为一个非传统的比较模型系统，以确定新的机制，抑制氧化应激 和铁性下垂。象海豹为与氧化应激相关的问题提供了一个特殊的模型系统 因为它们进化出了对氧化性组织损伤的显著抵抗力。在长达2小时的潜水中， 海象经历进行性和反复的组织缺血，但与缺血-再灌注相反， 在人类身上发生的伤害中，海象的组织受到保护，免受伤害。此外，我们的初步 数据证明培养的象海豹细胞对多种氧化应激源（例如，铁死亡 诱导剂），表明海象已经进化出细胞自主机制来抑制氧化应激， 损伤和铁性下垂。阐明了为海象细胞提供这种独特能力的机制， 防止氧化损伤提供了一个非凡的机会，发现意想不到的途径， 为衰老和与氧化应激相关的疾病提供见解，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， 研究已知的在培养的海象和人类细胞中的脂质过氧化和铁凋亡（目的1）。此外，委员会认为， 我们将进行全基因组合成致死筛选，系统地揭示铁凋亡的机制， 培养的海象细胞中的抗性（目的2）。这些研究的完成将提供第一个全基因组的 CRISPR-Cas9文库的大象海豹细胞，并将确定新的遗传修饰剂的ferroptosis，推出一项新的研究。 进入分子时代的新模型系统，可能会深入了解退行性疾病的病因， 并为新型治疗策略的开发提供基础。