PMID- 34473251 OWN - NLM STAT- MEDLINE DCOM- 20220110 LR - 20220219 IS - 1945-7170 (Electronic) IS - 0013-7227 (Print) IS - 0013-7227 (Linking) VI - 162 IP - 12 DP - 2021 Dec 1 TI - The Role of Mitochondria-Linked Fatty-Acid Uptake-Driven Adipogenesis in Graves Orbitopathy. LID - 10.1210/endocr/bqab188 [doi] LID - bqab188 AB - CONTEXT: Depot-specific expansion of orbital adipose tissue (OAT) in Graves orbitopathy (GO; an autoimmune condition producing proptosis, visual impairment and reduced quality of life) is associated with fatty acid (FA)-uptake-driven adipogenesis in preadipocytes/fibroblasts (PFs). OBJECTIVE: This work sought a role for mitochondria in OAT adipogenesis in GO. METHODS: Confluent PFs from healthy OAT (OAT-H), OAT from GO (OAT-GO) and white adipose tissue in culture medium compared with culture medium containing a mixed hormonal cocktail as adipogenic medium (ADM), or culture-medium containing FA-supplementation, oleate:palmitate:linoleate (45:30:25%) with/without different concentration of mitochondrial biosubstrate adenosine 5'-diphosphate/guanosine 5'-diphosphate (ADP/GDP), AICAR (adenosine analogue), or inhibitor oligomycin-A for 17 days. Main outcome measures included oil-red-O staining and foci count of differentiated adipocytes for in vitro adipogenesis, flow cytometry, relative quantitative polymerase chain reaction, MTS-assay/106 cells, total cellular-ATP detection kit, and Seahorse-XFe96-Analyzer for mitochondria and oxidative-phosphorylation (OXPHOS)/glycolysis-ATP production analysis. RESULTS: During early adipogenesis before adipocyte formation (days 0, 4, and7), we observed OAT-specific cellular ATP production via mitochondrial OXPHOS in PFs both from OAT-H and OAT-GO, and substantially disrupted OXPHOS-ATP/glycolysis-ATP production in PFs from OAT-GO, for example, a 40% reduction in OXPHOS-ATP and trend-increased glycolysis-ATP production on days 4 and 7 compared with day 0, which contrasted with the stable levels in OAT-H. FA supplementation in culture-medium triggered adipogenesis in PFs both from OAT-H and OAT-GO, which was substantially enhanced by 1-mM GDP reaching 7% to 18% of ADM adipogenesis. The FA-uptake-driven adipogenesis was diminished by oligomycin-A but unaffected by treatment with ADP or AICAR. Furthermore, we observed a significant positive correlation between FA-uptake-driven adipogenesis by GDP and the ratios of OXPHOS-ATP/glycolysis-ATP through adipogenesis of PFs from OAT-GO. CONCLUSION: Our study confirmed that FA uptake can drive OAT adipogenesis and revealed a fundamental role for mitochondria-OXPHOS in GO development, which provides potential for therapeutic interventions. CI - (c) The Author(s) 2021. Published by Oxford University Press on behalf of the Endocrine Society. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com. FAU - Zhang, Lei AU - Zhang L AUID- ORCID: 0000-0003-3536-8692 AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. FAU - Rai, Pavandeep AU - Rai P AD - Wellcome Centre for Mitochondrial Research, Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Newcastle, NE2 4HH, UK. FAU - Miwa, Satomi AU - Miwa S AD - Biosciences Institute, Newcastle University, Newcastle Upon Tyne, NE4 5PL, UK. FAU - Draman, Mohd Shazli AU - Draman MS AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. FAU - Rees, D Aled AU - Rees DA AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. FAU - Haridas, Anjana S AU - Haridas AS AD - Department of Ophthalmology, Cardiff & Vale University Health Board, Heath Park Hospital , Cardiff CF14 4XW, UK. FAU - Morris, Daniel S AU - Morris DS AD - Department of Ophthalmology, Cardiff & Vale University Health Board, Heath Park Hospital , Cardiff CF14 4XW, UK. FAU - Tee, Andrew R AU - Tee AR AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. FAU - Ludgate, Marian AU - Ludgate M AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. FAU - Turnbull, Doug M AU - Turnbull DM AD - Wellcome Centre for Mitochondrial Research, Translational and Clinical Research Institute, Faculty of Medical Sciences, Newcastle University, Newcastle, NE2 4HH, UK. FAU - Dayan, Colin M AU - Dayan CM AD - School of Medicine, Cardiff University, Heath Park Hospital, Cardiff, CF14 4XN, UK. LA - eng PT - Journal Article PL - United States TA - Endocrinology JT - Endocrinology JID - 0375040 RN - 0 (Fatty Acids) SB - IM MH - Adipocytes/metabolism MH - Adipogenesis/*physiology MH - Adipose Tissue/metabolism/pathology MH - Cell Differentiation MH - Cells, Cultured MH - Fatty Acids/*metabolism MH - Fibroblasts/metabolism/pathology MH - Graves Ophthalmopathy/*metabolism/pathology MH - Humans MH - Lipid Metabolism/physiology MH - Mitochondria/*physiology MH - Orbit MH - Oxidative Phosphorylation PMC - PMC8848742 OTO - NOTNLM OT - ATP production OT - Graves orbitopathy OT - adipogenesis OT - fatty acid uptake OT - mitochondria OXPHOS OT - orbital adipose tissue EDAT- 2021/09/03 06:00 MHDA- 2022/01/11 06:00 PMCR- 2021/09/02 CRDT- 2021/09/02 12:24 PHST- 2021/07/01 00:00 [received] PHST- 2021/09/03 06:00 [pubmed] PHST- 2022/01/11 06:00 [medline] PHST- 2021/09/02 12:24 [entrez] PHST- 2021/09/02 00:00 [pmc-release] AID - 6362764 [pii] AID - bqab188 [pii] AID - 10.1210/endocr/bqab188 [doi] PST - ppublish SO - Endocrinology. 2021 Dec 1;162(12):bqab188. doi: 10.1210/endocr/bqab188.