Molecular Mechanisms of FUNDC1-Mediated Mitophagy

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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.