Abstract
Parkin, an E3 ubiquitin ligase, plays an essential role in mitochondrial quality control. However, the mechanisms by which Parkin connects mitochondrial homeostasis with cellular metabolism in adipose tissue remain unclear. Here, we demonstrate that Park2 gene (encodes Parkin) deletion specifically from adipose tissue protects mice against high-fat diet and aging-induced obesity. Despite a mild reduction in mitophagy, mitochondrial DNA content and mitochondrial function are increased in Park2 deficient white adipocytes. Moreover, Park2 gene deletion elevates mitochondrial biogenesis by increasing Pgc1α protein stability through mitochondrial superoxide-activated NAD(P)H quinone dehydrogenase 1 (Nqo1). Both in vitro and in vivo studies show that Nqo1 overexpression elevates Pgc1α protein level and mitochondrial DNA content and enhances mitochondrial activity in mouse and human adipocytes. Taken together, our findings indicate that Parkin regulates mitochondrial homeostasis by balancing mitophagy and Pgc1α-mediated mitochondrial biogenesis in white adipocytes, suggesting a potential therapeutic target in adipocytes to combat obesity and obesity-associated disorders.
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Introduction
Mitochondrial dysfunction contributes to the pathogenesis of metabolic disorders such as obesity1. Mitochondria are highly dynamic organelles that play essential roles in energy metabolism, heat production, generation of oxygen radicals, calcium signaling, and apoptosis2,3,4,5,6. To maintain metabolic health, mitochondrial selective autophagy (mitophagy), mitochondrial biogenesis, and various mitochondrial protein degradation processes coordinately control the quality of mitochondria6,7.
Mitophagy involves the recognition of damaged mitochondria by the autophagosome through microtubule-associated protein 1 light chain 3 (LC3) adapters in ubiquitin-dependent and independent mechanisms8,9. PTEN-induced putative kinase 1 (Pink1, encoded by Park6) and E3 ubiquitin-protein ligase Parkin are known to function in ubiquitin-dependent mitophagy, while other proteins such as TBC1 domain family member 15 (Tbc1d15) and Nip3-like protein X (Nix) contribute to ubiquitin-independent mitophagy9,10,11,12.
Emerging evidence has shown that low mitochondrial content and activity contribute to adipocyte hypertrophy and metabolic disorders13,14,15,S1 for a list of the primers used.
RNA sequencing and analysis
A homogenous portion of frozen adipose samples were homogenized using a tissue homogenizer in Trizol. RNA was isolated using the Qiagen RNeasy Mini QIAcube Kit following the manufacturer’s instructions. Isolated RNA was checked for concentration using a NanoDrop and purity using the Agilent TapeStation. Only samples with RIN > 7.0 were used. Libraries were prepared using the KAPA mRNA HyperPrep Kit following the manufacturer’s instructions. The resulting libraries were combined into two pools and sequenced on an Illumina HiSeq 3000 within the UCLA TCGB core facility following in-house established protocols. Raw reads were checked for quality using FastQC, aligned to the Mus musculus GRCm38, and then counted using Mus musculus GRCm38 version 97. Alignment and counting occurred using Rsubread v 2.4.263. Raw counts were then analyzed for differential gene expression using the DESeq2 v1.30.064.
Statistics
Values presented are expressed as means ± SEM unless otherwise indicated. Statistical analyses were performed using Student’s t test when comparing two groups of samples or Two-way analysis of variance (ANOVA) for identification of significance within and between groups using GraphPad Prism9.4.1 (GraphPad Software). Significance was set a priori at P < 0.05. For RNA Sequencing analysis, the significance was set as FDR < 0.05.
Reporting summary
Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.
Data availability
Source data are provided as a Source Data file. METSIM adipose array data are available from in the Gene Expression Omnibus database under accession code GSE70353, and additional clinical data were obtained from previously published papers60,65. The mouse RNA-seq results from eWAT of HFD-fed ParkinAdi vs. Control f/f mice and AAV8-Nqo1 vs. AAV8-Control injected eWAT have been deposited in the Gene Expression Omnibus database under accession code GSE207496. Human PARK2 gene (encodes Parkin protein) association is available from PhenoScanner v2 (http://www.phenoscanner.medschl.cam.ac.uk). The authors declare that all data supporting the findings of this study are available within the paper and its Supplementary Information files.
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Acknowledgements
We would like to thank Dr. Ted M. Dawson at Johns Hopkins University for providing us Parkin flox mice. We thank undergraduate students Julia Zhou at the University of California San Diego for analyzing the adipocyte size. We thank Wenjuan Ren for immunoblotting and Calvin Pan for HMDP data analysis. Z.Z. was supported by an NIH grant (DK125354). A.L.H. was supported by NIH grants (DK109724, P30DK063491). A.J.L. was supported by NIH grants (U54 DK120342 and DK117850). T.M.M. was supported by the UCLA Intercampus Medical Genetics Training Program (T32GM008243), NRSA predoctoral fellowship (F31DK108657), Carl V. Gisolfi Memorial Research grant from the American College of Sports Medicine, and a predoctoral graduate student award from the Dornsife College at the University of Southern California. D.M.W. is supported by an EMBO long-term fellowship ALTF 828-2021.
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Z.Z., T.M.M., and A.L.H conceived and designed the experiments. Z.Z., T.M.M., A.R.S., and X.Z. performed all in vivo studies. Z.Z., T.M.M., X.Z., A.R.S., Y.C., A.M., and P.S. conducted the sample collection and subsequent experimental analysis. B.L.C., and T.Q.d.A.V. performed indirect calorimetry studies. Z.Z. and T.M.M. performed all adipocyte culture studies and mtDNA content analysis. L.C. performed all AAV injection studies. D.M.W., J.N., M.S. conducted studies related to mitochondrial respirometry and confocal microscopy. Z.Z., T.M.M., L.C., X.Z., A.R.S., Y.C., B.L.C., A.M., P.S. T.Q.d.A.V., M.L., O.S.S., A.L.H, and A.J.L. analyzed the data. T.M.M., L.C., A.L.H, A.J.L., and Z.Z. wrote the paper.
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Moore, T.M., Cheng, L., Wolf, D.M. et al. Parkin regulates adiposity by coordinating mitophagy with mitochondrial biogenesis in white adipocytes. Nat Commun 13, 6661 (2022). https://doi.org/10.1038/s41467-022-34468-2
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DOI: https://doi.org/10.1038/s41467-022-34468-2
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