Molecular Mechanisms of FUNDC1-Mediated Mitophagy

FUNDC1介导的线粒体自噬的分子机制

• 批准号: 10625338
• 负责人: Jose M Delgado
• 金额: $ 4.77万
• 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/10625338
• 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; Cytoplasm; Data; Development; Disease; Ensure; Event; Experimental Designs; Future; Genes; Genetic; Genetic Screening; Goals; Homeostasis; Human Pathology; Hypoxia; Laboratories; 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; Postdoctoral Fellow; Proteins; Receptor Activation; Regulation; Reperfusion Injury; Reporter; Research; Research Personnel; Research Project Grants; Role; Science; Site; Stress; Technical Expertise; Therapeutic; Training; career; college; experience; experimental study; genome-wide; high throughput technology; insight; interest; mutant; novel; programs; protein oligomer; 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，有选择地靶向和降解细胞质含量。 自噬靶标的识别是细胞稳态的关键方面，并由选择性介导 自噬受体。在表征可溶性自噬受体方面取得了巨大进展。 然而，最近发现的膜包裹的受体Fun14域14的机制1 （Fundc1）仍然未知。 Fundc1位于线粒体外膜上介导 线粒体的自噬周转，称为线粒体。基金C1介导的线粒体被诱导 病理和发展计划。例如，以前的研究证明了生理 Fundc1介导的线粒体在与缺氧相关的病理中的作用，包括癌症和缺血 - 再灌注 受伤。初步结果表明，基金C1介导的线索与其他形式的分子不同 线粒体。该提案的总体目的是阐明缺氧诱导的分子机制 选择性自噬途径通过Fundc1函数的表征。在AIM 1中，域分析 将执行方法以识别缺氧引起的基金C1的功能重要区域 线粒体。这些研究将剖析能够激活Fundc1的潜在激活事件。在AIM 2中， 用于高通量遗传筛查的CRISPR-CAS9技术将用于识别Fundc1的调节剂 周转。调节基金C1介导的线粒体的新型遗传因素将以其特征为特征 功能。综上所述，提出的实验将为阐明缺氧的机制提供见识。 诱导的选择性自噬并扩展了与缺氧相关的人类自噬的假定治疗靶标 病理。通过拟议的培训计划，我将增强进步我的科学研究所需的技能 职业包括实验室技术技能，实验设计，科学沟通和指导。 达特茅斯学院和我的指导团队有能力确保我的研究项目的成功 成为博士后研究员的学术生涯的下一步。