Molecular Mechanisms of FUNDC1-Mediated Mitophagy

摘要 自噬是溶酶体介导的选择性靶向和降解细胞质内容物的途径。 自噬靶点的鉴定是细胞稳态的关键方面，并且由选择性自噬介导。 自噬受体在表征可溶性自噬受体方面已经取得了巨大的进展。 然而，最近发现的膜包埋受体Fun 14结构域的机制， （FUNDC 1）仍然鲜为人知。位于线粒体外膜，FUNDC 1介导 线粒体的自噬周转，称为线粒体自噬。FUNDC 1介导的线粒体自噬在多种细胞中诱导 病理学和发展计划。例如，以前的研究表明， FUNDC 1介导的线粒体自噬在缺氧相关病理学（包括癌症和缺血再灌注）中的作用 损伤初步结果表明，FUNDC 1介导的线粒体自噬在分子上不同于其他形式的线粒体自噬。 线粒体自噬这一建议的总体目标是阐明缺氧诱导的细胞凋亡的分子机制。 选择性自噬途径通过表征FUNDC 1功能。在目标1中，域分析 方法将执行，以确定功能上重要的区域FUNDC 1所需的缺氧诱导 线粒体自噬这些研究将剖析能够激活FUNDC 1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FUNDC 1的调节剂 周转调节FUNDC 1介导的线粒体自噬的新型遗传因子将表征其特征 功能总之，拟议的实验将提供深入了解阐明缺氧的机制- 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 病理学通过建议的培训计划，我将提高我的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，科学交流和指导。 达特茅斯学院和我的导师团队装备精良，以确保我的研究项目的成功， 作为博士后研究员，我的学术生涯的下一步。

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.