Molecular Mechanisms of FUNDC1-Mediated Mitophagy

FUNDC1介导的线粒体自噬的分子机制

• 批准号: 10461452
• 负责人: Jose M Delgado
• 金额: $ 4.68万
• 依托单位: DARTMOUTH COLLEGE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10461452
• 关键词: Adenoviruses; Autophagocytosis; BCL2/Adenovirus E1B 19kd Interacting Protein 3-Like; BNIP3L gene; Bypass; CRISPR screen; CRISPR/Cas technology; Cardiovascular Diseases; Cells; Communication; Data; Development; Disease; Ensure; Event; Experimental Designs; Fibrinogen; Future; Genes; Genetic; Genetic Screening; Goals; Homeostasis; Human Pathology; Hypoxia; Laboratories; Lipids; Lysosomes; Malignant Neoplasms; Mediating; Membrane; Mentorship; Mitochondria; Molecular; Monitor; Mutate; Organelles; Outcomes Research; Outer Mitochondrial Membrane; Oxygen; Oxygen Consumption; PINK1 gene; Parkin; Pathology; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological; Proteins; Receptor Activation; Regulation; Reperfusion Injury; Reporter; Research; Research Personnel; Research Project Grants; Role; Science; Site; Stress; Technical Expertise; Therapeutic; Training; base; career; college; experience; experimental study; genome-wide; high throughput technology; insight; interest; mutant; novel; programs; receptor; response; skills; success; therapeutic target; tool; ubiquitin-protein ligase

ABSTRACT Autophagy is a lysosome-mediated pathway that selectively targets and degrades cytoplasmic content. Identification of autophagy targets is a critical facet of cellular homeostasis and is mediated by selective autophagy receptors. Immense progress has been made in characterizing soluble autophagy receptors. However, mechanisms of recently discovered membrane-embedded receptor Fun14 domain-containing 1 (FUNDC1) remains widely unknown. Located on the outer mitochondrial membrane, FUNDC1 mediates autophagic turnover of mitochondria, termed mitophagy. FUNDC1-mediated mitophagy is induced in diverse pathologies and developmental programs. For example, previous studies have demonstrated a physiological role for FUNDC1-mediated mitophagy in hypoxia-related pathologies, including cancer and ischemia-reperfusion injury. Preliminary results suggest that FUNDC1-mediated mitophagy is molecularly distinct from other forms of mitophagy. The overall objective of this proposal is to elucidate the molecular mechanisms of hypoxia-induced selective autophagy pathways through the characterization of FUNDC1 function. In Aim 1, a domain analysis approach will be performed to identify functionally important regions of FUNDC1 required for hypoxia-induced mitophagy. These studies will dissect potential activation events that enable activation of FUNDC1. In Aim 2, CRISPR-Cas9 technology for high-throughput genetic screens will be used to identify modulators of FUNDC1 turnover. Novel genetic factors that modulate FUNDC1-mediated mitophagy will be characterized for their function. Taken together, the proposed experiments will provide insight to elucidate mechanisms of hypoxia- induced selective autophagy and expand putative therapeutic targets for autophagy in hypoxic-related human pathologies. With the proposed training plan, I will enhance the skills needed to progress my scientific research career including laboratory technical skills, experimental design, science communication, and mentorship. Dartmouth College and my mentorship team are well-equipped to ensure success of my research project and progression to the next step of my academic career as a postdoctoral researcher.

抽象的 自噬是一种溶酶体介导的途径，选择性地靶向并降解细胞质内容。 自噬靶标的识别是细胞稳态的一个关键方面，由选择性介导 自噬受体。在表征可溶性自噬受体方面已经取得了巨大进展。 然而，最近发现的膜嵌入受体 Fun14 结构域的机制 1 (FUNDC1) 仍然广为人知。 FUNDC1 位于线粒体外膜，介导 线粒体的自噬周转，称为线粒体自噬。 FUNDC1 介导的线粒体自噬在多种细胞中被诱导 病理学和发育计划。例如，之前的研究已经证明了生理学 FUNDC1 介导的线粒体自噬在缺氧相关病理（包括癌症和缺血再灌注）中的作用 受伤。初步结果表明 FUNDC1 介导的线粒体自噬在分子上与其他形式的线粒体自噬不同。 线粒体自噬。该提案的总体目标是阐明缺氧诱导的分子机制 通过 FUNDC1 功能表征选择性自噬途径。在目标 1 中，域分析 将进行方法来识别缺氧诱导所需的 FUNDC1 功能重要区域 线粒体自噬。这些研究将剖析能够激活 FUNDC1 的潜在激活事件。在目标 2 中， 用于高通量遗传筛选的 CRISPR-Cas9 技术将用于鉴定 FUNDC1 的调节剂 周转。调节 FUNDC1 介导的线粒体自噬的新遗传因子将被表征为 功能。总而言之，所提出的实验将为阐明缺氧机制提供见解。 诱导选择性自噬并扩大缺氧相关人类自噬的假定治疗靶点 病理学。通过拟议的培训计划，我将提高开展科学研究所需的技能 职业生涯包括实验室技术技能、实验设计、科学传播和指导。 达特茅斯学院和我的导师团队装备精良，可以确保我的研究项目取得成功， 作为博士后研究员，我将进入学术生涯的下一步。