Stress tolerant annual killifish: a new model for the cellular stress response

耐压一年生鳉鱼：细胞应激反应的新模型

• 批准号: 10570347
• 负责人: Claire L Riggs
• 金额: $ 12.47万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-22 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10570347
• 关键词: Anoxia; Automobile Driving; Biogenesis; Biological Assay; Biology; Career Mobility; Cell Survival; Cell model; Cell physiology; Cells; Cellular Stress; Cellular biology; Cytoplasm; Data; Disease; Embryo; Environment; Equilibrium; Female; Fishes; Fostering; G3BP1 gene; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Genomics; Goals; Hospitals; Human; Human Cell Line; Killifishes; Life; Membrane; Mentors; Messenger RNA; MicroRNAs; Mitochondria; Mitochondrial Diseases; Modeling; Mutation; Myocardial Infarction; Neurodegenerative Disorders; Nuclear; Organ Transplantation; Organelles; Organism; Oxygen; Phase; Protein Biosynthesis; Protein Synthesis Inhibition; Proteins; RNA; Regulator Genes; Research; Research Personnel; Resources; Scientist; Stress; Stroke; Study models; Testing; Therapeutic; Training; Transfer RNA; Translational Regulation; Translational Repression; Translations; Untranslated RNA; Up-Regulation; Venezuela; Vertebrates; Woman; Work; biological adaptation to stress; career development; higher education; human disease; innovation; insight; medical schools; metabolic depression; novel; novel strategies; novel therapeutics; overexpression; prevent; programs; response; ribosome profiling; selective expression; sodium arsenite; stress granule; stress tolerance; stressor; success

Project Summary/Abstract When faced with environmental stress, cells and organisms must conserve resources by decreasing global translation and simultaneously upregulating stress-responsive genes. How cells do this has eluded researchers yet promises insight into basic cell biology and novel therapeutics. This proposal takes an innovative approach to the problem by developing a naturally stress-tolerant fish as a model for the cellular stress response and using it to examine the contribution of membraneless organelles (in particular stress granules (SGs)) and small non-coding RNAs (ncRNAs) to gene expression regulation and stress tolerance. SGs are highly conserved stress-induced assemblies of RNA and protein which may regulate gene expression by selective sequestration of RNAs and proteins, however their function remains unknown. Small ncRNAs, such as microRNAs, can regulate the gene expression of particular mRNAs by preventing their translation into protein, yet identifying adaptive ncRNAs that function in the stress response remains challenging. The annual killifish Austrofundulus limnaeus inhabits temporary ponds in Venezuela that expose developing fish embryos to harsh environmental conditions, including anoxia. A. limnaeus embryos can survive over 100 days without oxygen and cells derived from A. limnaeus embryos can survive two weeks without oxygen. A. limnaeus display unique SG and ncRNA biology. Our preliminary data show that A. limnaeus cells treated with sodium arsenite do not form SGs, but still shut down translation. This surprising finding suggests that SG avoidance may be adaptive and support anoxia tolerance in killifish cells. Small ncRNA studies in anoxia-tolerant vertebrates identified mitochondria-derived small ncRNAs (mitosRNAs) as putative regulators of gene expression driving anoxia-induced metabolic depression in A. limnaeus embryos. These mitosRNAs may therefore underpin organismal anoxia tolerance and represent a novel stress response mechanism. This proposal will address the following questions: 1. How do killifish cells decouple translation inhibition from stress granule formation?; 2. How do mitosRNAs regulate protein synthesis under stress?; and 3. How do SG avoidance and mitosRNA expression support anoxia tolerance? Understanding the mechanism and consequences of these cellular phenomena will offer unique insight into fundamental cell biology that may inform treatments for heart attack and stroke, as well as mitochondrial and neurodegenerative diseases. This proposal will facilitate the career development and transition to independence of a female scientist committed to promoting and supporting diversity in STEM. The mentored phase will be conducted in the rich training environment of Brigham and Women’s Hospital/Harvard Medical School to facilitate her career development. Transitioning to independence will poise her to advance inclusive excellence in higher education and STEM, including fostering the success of diverse trainees her in lab.

项目总结/摘要 当面临环境压力时，细胞和生物体必须通过减少全球变暖来保护资源。 翻译并同时上调应激反应基因。细胞是如何做到这一点的， 研究人员还承诺深入了解基础细胞生物学和新的治疗方法。这项建议需要一个 创新的方法来解决这个问题，通过开发一个自然的压力耐受鱼作为模型的细胞 应激反应，并用它来检查无膜细胞器（特别是应激）的贡献 颗粒（SGs））和小的非编码RNA（ncRNA）对基因表达调控和胁迫耐受性的影响。 SGs是高度保守的胁迫诱导的RNA和蛋白质组装体，可能调节基因表达 通过RNA和蛋白质的选择性螯合，然而它们的功能仍然未知。小ncRNA， 如microRNA，可以通过阻止它们翻译成 然而，鉴定在应激反应中起作用的适应性ncRNA仍然具有挑战性。年度 在委内瑞拉的临时池塘里，一条南极鱼栖息在那里，露出了正在发育的鱼胚胎。 恶劣的环境条件，包括缺氧。A. Limnaeus胚胎可以存活超过100天， 氧和细胞来源于A. Limnaeus胚胎可以在没有氧气的情况下存活两周。A.利姆奈 显示独特的SG和ncRNA生物学。初步数据表明，A.用钠处理的limnaeus细胞 亚砷酸盐不形成SGs，但仍然关闭翻译。这一令人惊讶的发现表明，SG回避 可能是适应性的，并支持缺氧耐受性的细胞。耐缺氧小鼠中的小ncRNA研究 脊椎动物鉴定出了来自于哺乳动物的小ncRNA（mitosRNA）作为基因的假定调节因子， 表达驱动缺氧诱导的A. limnaeus胚胎这些线粒体RNA可能 因此支撑了生物体的缺氧耐受性，并代表了一种新型的应激反应机制。这 建议将解决以下问题：1. Killifish细胞如何将翻译抑制与压力分离 颗粒形成？2.应激条件下线粒体RNA如何调节蛋白质合成和3. SG怎么样 回避和mitosRNA表达支持缺氧耐受？了解机制和 这些细胞现象的结果将提供对基本细胞生物学的独特见解， 为心脏病发作和中风以及线粒体和神经退行性疾病的治疗提供信息。这 该提案将促进致力于实现职业发展和向独立过渡的女科学家 促进和支持STEM领域的多样性。指导阶段将在丰富的培训中进行 布里格姆妇女医院/哈佛医学院的环境，以促进她的职业发展。 过渡到独立将使她能够在高等教育和STEM领域推进包容性卓越， 包括在实验室中培养不同学员的成功。