PT  - JOURNAL ARTICLE
AU  - Simone Cristina da Silva Rosa
AU  - Matthew D. Martens
AU  - Jared T. Field
AU  - Lucas Nguyen
AU  - Stephanie M. Kereliuk
AU  - Yan Hai
AU  - Donald Chapman
AU  - William Diehl-Jones
AU  - Michel Aliani
AU  - Adrian R. West
AU  - James Thliveris
AU  - Saeid Ghavami
AU  - Christof Rampitsch
AU  - Vernon W. Dolinsky
AU  - Joseph W. Gordon
TI  - Nix induced mitochondrial fission, mitophagy, and myocyte insulin resistance are abrogated by PKA phosphorylation
AID  - 10.1101/825828
DP  - 2019 Jan 01
TA  - bioRxiv
PG  - 825828
4099  - http://biorxiv.org/content/early/2019/11/04/825828.short
4100  - http://biorxiv.org/content/early/2019/11/04/825828.full
AB  - Lipotoxicity is a form of cellular stress caused by the accumulation of lipids resulting in mitochondrial dysfunction and insulin resistance in muscle. Previously, we demonstrated that the mitophagy receptor Nix is responsive to lipotoxicity and accumulates in response to diacylglycerols induced by high-fat (HF) feeding. In addition, previous studies have implicated autophagy and mitophagy in muscle insulin sensitivity. To provide a better understanding of these observations, we undertook gene expression array and shot-gun metabolomics studies in soleus muscle from rodents on an HF diet. Interestingly, we observed a modest reduction in several autophagy-related genes including Beclin-1, ATG3, and −5. Moreover, we observed alterations in the fatty acyl composition of cardiolipins and phosphatidic acids. Given the previously reported roles of these phospholipids and Nix in mitochondrial dynamics, we investigated aberrant mitochondrial fission and turn-over as a mechanism of myocyte insulin resistance. In a series of gain-of-function and loss-of-function experiments in rodent and human myotubes, we demonstrate that Nix accumulation triggers mitochondrial depolarization, fragmentation, calcium-dependent activation of DRP1, and mitophagy. In addition, Nix-induced mitochondrial fission leads to myotube insulin resistance through activation of mTOR-p70S6 kinase inhibition of IRS1, which is contingent on phosphatidic acids and Rheb1. Finally, we demonstrate that Nix-induced mitophagy and insulin resistance can be reversed by direct phosphorylation of Nix by PKA, leading to the translocation of Nix from the mitochondria and sarcoplasmic reticulum to the cytosol. These findings provide insight into the role of Nix-induced mitophagy and myocyte insulin resistance during an overfed state when overall autophagy-related gene expression is reduced. Furthermore, our data suggests a mechanism by which exercise or pharmacological activation of PKA may overcome myocyte insulin resistance.