{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T20:05:12Z","timestamp":1790193912241,"version":"4.1.0"},"posted":{"date-parts":[[2026,9,18]]},"group-title":"Immunology","reference-count":82,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T00:00:00Z","timestamp":1789689600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/www.biorxiv.org\/about\/FAQ#license"}],"funder":[{"name":"NIH","award":["GM-147713"],"award-info":[{"award-number":["GM-147713"]}]},{"name":"NIH","award":["GM-139763"],"award-info":[{"award-number":["GM-139763"]}]},{"name":"NIH","award":["GM-137921"],"award-info":[{"award-number":["GM-137921"]}]},{"name":"NIH","award":["GM-144103"],"award-info":[{"award-number":["GM-144103"]}]},{"name":"NIH","award":["S10OD028479"],"award-info":[{"award-number":["S10OD028479"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2026,9,18]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:p>Invariant natural killer T (iNKT) cells require mitochondrial metabolism for terminal effector development. We found iNKT cells express elevated levels of proteins regulating mitochondrial membrane dynamics, and identified Opa1, but not Drp1, as selectively required for iNKT cell differentiation. OPA1 deficiency disrupted mitochondrial cristae organization, reduced mitochondrial membrane potential, increased mitochondrial mass, and altered calcium homeostasis in iNKT cells. Bulk and single-cell transcriptomic analyses revealed impaired TCR-responsive gene expression, activation of mitochondrial stress adaptation and integrated stress response, enhanced glycolysis, and retention of immature differentiation features. SCENITH analysis demonstrated increased glycolytic dependence; LDHA became required in Opa1-deficient iNKT cells while dispensable for normal iNKT development, indicating compensatory glycolytic adaptation. EGTA in thymic organ culture partially restored NKT1 marker expression in Opa1-deficient cells. Co-deletion of Drp1 improved mitochondrial morphology, TCR signaling, stress and metabolic adaptation, and partially rescued iNKT cell differentiation, demonstrating that balanced mitochondrial dynamics coordinate mitochondrial function and terminal effector development.<\/jats:p>","DOI":"10.64898\/2026.09.12.751163","type":"posted-content","created":{"date-parts":[[2026,9,18]],"date-time":"2026-09-18T22:10:13Z","timestamp":1789769413000},"source":"Crossref","is-referenced-by-count":0,"title":["OPA1-dependent mitochondrial remodeling coordinates TCR signaling and metabolic adaptation during iNKT cell differentiation"],"prefix":"10.64898","author":[{"given":"Sophia P. M.","family":"Sok","sequence":"first","affiliation":[{"name":"Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianan","family":"Cheng","sequence":"additional","affiliation":[{"name":"Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rufei","family":"Lu","sequence":"additional","affiliation":[{"name":"Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"},{"name":"Department of Pathology, The University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7033-7010","authenticated-orcid":false,"suffix":"Jr","given":"Tommy L","family":"Lewis","sequence":"additional","affiliation":[{"name":"Aging and Metabolism Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"},{"name":"Department of Biochemistry & Physiology, University of Oklahoma Health Campus, Oklahoma City, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hiromi","family":"Sesaki","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Renata O","family":"Pereira","sequence":"additional","affiliation":[{"name":"Department of Internal Medicine, Division of Endocrinology and Metabolism, Fraternal Order of Eagles Diabetes Research Center, University of Iowa, 169 Newton Road, 4322 PBDB, Iowa City, IA, 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kai","family":"Sun","sequence":"additional","affiliation":[{"name":"Center for Metabolic and Degenerative Diseases, The Brown Foundation Institute of Molecular Medicine for the Prevention of Human Diseases, University of Texas Health Science Center at Houston, Houston, Texas, USA; Graduate Program in Biochemistry and Cellular Biology, Graduate School of Biomedical Sciences, University of Texas Health Science Center at Houston, Houston, Texas, USA. Electronic address:"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victoria","family":"Sun","sequence":"additional","affiliation":[{"name":"Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6392-4954","authenticated-orcid":false,"given":"Meng","family":"Zhao","sequence":"additional","affiliation":[{"name":"Arthritis and Clinical Immunology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, Oklahoma, USA"},{"name":"Department of Microbiology and Immunology, Stephenson Cancer Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"54368","reference":[{"key":"2026092312300928000_2026.09.12.751163v1.1","doi-asserted-by":"publisher","DOI":"10.1038\/s41577-020-0345-y"},{"key":"2026092312300928000_2026.09.12.751163v1.2","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1038\/s41577-025-01219-5","article-title":"Metabolic control of innate-like T cells","volume":"26","year":"2026","journal-title":"Nat Rev Immunol"},{"key":"2026092312300928000_2026.09.12.751163v1.3","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.0803394"},{"key":"2026092312300928000_2026.09.12.751163v1.4","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0812255106"},{"key":"2026092312300928000_2026.09.12.751163v1.5","doi-asserted-by":"crossref","first-page":"516","DOI":"10.1002\/eji.201646343","article-title":"mTORC2 regulates multiple aspects of NKT-cell development and function","volume":"47","year":"2017","journal-title":"Eur J Immunol"},{"key":"2026092312300928000_2026.09.12.751163v1.6","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1401985"},{"key":"2026092312300928000_2026.09.12.751163v1.7","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1400769"},{"key":"2026092312300928000_2026.09.12.751163v1.8","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1402042"},{"key":"2026092312300928000_2026.09.12.751163v1.9","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1315435111"},{"key":"2026092312300928000_2026.09.12.751163v1.10","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1901376116"},{"key":"2026092312300928000_2026.09.12.751163v1.11","doi-asserted-by":"crossref","unstructured":"Weng X , Kumar A , Cao L , He Y , Morgun E , Visvabharathy L , Zhao J , Sena LA , Weinberg SE , Chandel NS and Wang CR . Mitochondrial metabolism is essential for invariant natural killer T cell development and function. Proc Natl Acad Sci U S A. 2021;118.","DOI":"10.1073\/pnas.2021385118"},{"key":"2026092312300928000_2026.09.12.751163v1.12","doi-asserted-by":"crossref","unstructured":"Sok SPM , Cheng J , Strucinska K , Popescu NI , Wu L , Ke Q , Kiosses WB , Stanford D , Freeman WM , Matsuzaki S , Lewis TL, Jr. and Zhao M . P-glycoprotein expression skews mitochondrial dye measurements in T cells. Front Immunol. 2025;16:1560104.","DOI":"10.3389\/fimmu.2025.1560104"},{"key":"2026092312300928000_2026.09.12.751163v1.13","first-page":"e107901","article-title":"Calcium signals regulate the functional differentiation of thymic iNKT cells","volume":"40","year":"2021","journal-title":"EMBO J"},{"key":"2026092312300928000_2026.09.12.751163v1.14","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-020-0210-7"},{"key":"2026092312300928000_2026.09.12.751163v1.15","doi-asserted-by":"publisher","DOI":"10.1038\/s41580-024-00785-1"},{"key":"2026092312300928000_2026.09.12.751163v1.16","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2008.11.046"},{"key":"2026092312300928000_2026.09.12.751163v1.17","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2013.08.032"},{"key":"2026092312300928000_2026.09.12.751163v1.18","doi-asserted-by":"publisher","DOI":"10.1038\/nrm3412"},{"key":"2026092312300928000_2026.09.12.751163v1.19","doi-asserted-by":"crossref","unstructured":"Chandel NS . Mitochondria as signaling organelles. BMC Biol. 2014;12:34.","DOI":"10.1186\/1741-7007-12-34"},{"key":"2026092312300928000_2026.09.12.751163v1.20","doi-asserted-by":"publisher","DOI":"10.1038\/s41418-021-00747-6"},{"key":"2026092312300928000_2026.09.12.751163v1.21","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2018.11.018"},{"key":"2026092312300928000_2026.09.12.751163v1.22","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2016.05.035"},{"key":"2026092312300928000_2026.09.12.751163v1.23","doi-asserted-by":"crossref","unstructured":"Willett BAS , Thompson SB , Chen V , Dareshouri A , Jackson CL , Brunetti T , D\u2019Alessandro A , Klarquist J , Nemkov T and Kedl RM . Mitochondrial protein OPA1 is required for the expansion of effector CD8 T cells. Cell Rep. 2025;44:115610.","DOI":"10.1016\/j.celrep.2025.115610"},{"key":"2026092312300928000_2026.09.12.751163v1.24","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-022-05264-1"},{"key":"2026092312300928000_2026.09.12.751163v1.25","doi-asserted-by":"publisher","DOI":"10.1002\/dvg.22050"},{"key":"2026092312300928000_2026.09.12.751163v1.26","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.0407043101"},{"key":"2026092312300928000_2026.09.12.751163v1.27","doi-asserted-by":"publisher","DOI":"10.1016\/j.cell.2006.06.025"},{"key":"2026092312300928000_2026.09.12.751163v1.28","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.12.8.2245"},{"key":"2026092312300928000_2026.09.12.751163v1.29","doi-asserted-by":"publisher","DOI":"10.15252\/emmm.201506159"},{"key":"2026092312300928000_2026.09.12.751163v1.30","doi-asserted-by":"publisher","DOI":"10.1038\/embor.2010.214"},{"key":"2026092312300928000_2026.09.12.751163v1.31","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E07-12-1205"},{"key":"2026092312300928000_2026.09.12.751163v1.32","doi-asserted-by":"publisher","DOI":"10.1128\/MCB.01177-13"},{"key":"2026092312300928000_2026.09.12.751163v1.33","doi-asserted-by":"crossref","unstructured":"Chen Z , Lei C , Wang C , Li N , Srivastava M , Tang M , Zhang H , Choi JM , Jung SY , Qin J and Chen J . Global phosphoproteomic analysis reveals ARMC10 as an AMPK substrate that regulates mitochondrial dynamics. Nat Commun. 2019;10:104.","DOI":"10.1038\/s41467-018-08004-0"},{"key":"2026092312300928000_2026.09.12.751163v1.34","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2015.03.004"},{"key":"2026092312300928000_2026.09.12.751163v1.35","doi-asserted-by":"publisher","DOI":"10.1016\/j.febslet.2007.06.052"},{"key":"2026092312300928000_2026.09.12.751163v1.36","doi-asserted-by":"crossref","unstructured":"Raynor JL , Liu C , Dhungana Y , Guy C , Chapman NM , Shi H , Neale G , Sesaki H and Chi H . Hippo\/Mst signaling coordinates cellular quiescence with terminal maturation in iNKT cell development and fate decisions. J Exp Med. 2020;217.","DOI":"10.1084\/jem.20191157"},{"key":"2026092312300928000_2026.09.12.751163v1.37","doi-asserted-by":"crossref","first-page":"1376","DOI":"10.4049\/jimmunol.2200809","article-title":"TGF-beta Promotes the Postselection Thymic Development and Peripheral Function of IFN-gamma-Producing Invariant NKT cells","volume":"211","year":"2023","journal-title":"J Immunol"},{"key":"2026092312300928000_2026.09.12.751163v1.38","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbabio.2010.01.005"},{"key":"2026092312300928000_2026.09.12.751163v1.39","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.137.3.633"},{"key":"2026092312300928000_2026.09.12.751163v1.40","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-20073-8"},{"key":"2026092312300928000_2026.09.12.751163v1.41","doi-asserted-by":"publisher","DOI":"10.1101\/gr.108696.110"},{"key":"2026092312300928000_2026.09.12.751163v1.42","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(03)00105-9"},{"key":"2026092312300928000_2026.09.12.751163v1.43","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.201702058"},{"key":"2026092312300928000_2026.09.12.751163v1.44","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-020-2078-2"},{"key":"2026092312300928000_2026.09.12.751163v1.45","doi-asserted-by":"crossref","unstructured":"Zhao M and Wu L . Beyond subsets: single cell transcriptomics reveals multidimensional regulation of iNKT cells cross tissues and species. Front Immunol. 2026;17:1874403.","DOI":"10.3389\/fimmu.2026.1874403"},{"key":"2026092312300928000_2026.09.12.751163v1.46","doi-asserted-by":"publisher","DOI":"10.1016\/j.cub.2016.06.002"},{"key":"2026092312300928000_2026.09.12.751163v1.47","doi-asserted-by":"publisher","DOI":"10.1038\/nrm.2017.66"},{"key":"2026092312300928000_2026.09.12.751163v1.48","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200906084"},{"key":"2026092312300928000_2026.09.12.751163v1.49","doi-asserted-by":"publisher","DOI":"10.1038\/cr.2018.16"},{"key":"2026092312300928000_2026.09.12.751163v1.50","doi-asserted-by":"publisher","DOI":"10.1038\/sj.emboj.7601249"},{"key":"2026092312300928000_2026.09.12.751163v1.51","doi-asserted-by":"publisher","DOI":"10.1038\/sj.embor.7400790"},{"key":"2026092312300928000_2026.09.12.751163v1.52","doi-asserted-by":"publisher","DOI":"10.1038\/nature18302"},{"key":"2026092312300928000_2026.09.12.751163v1.53","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2020.02.017"},{"key":"2026092312300928000_2026.09.12.751163v1.54","doi-asserted-by":"publisher","DOI":"10.1038\/ni.3437"},{"key":"2026092312300928000_2026.09.12.751163v1.55","doi-asserted-by":"publisher","DOI":"10.1084\/jem.20110308"},{"key":"2026092312300928000_2026.09.12.751163v1.56","doi-asserted-by":"publisher","DOI":"10.1038\/sj.onc.1204386"},{"key":"2026092312300928000_2026.09.12.751163v1.57","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M709887200"},{"key":"2026092312300928000_2026.09.12.751163v1.58","doi-asserted-by":"publisher","DOI":"10.1038\/ni.2055"},{"key":"2026092312300928000_2026.09.12.751163v1.59","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2013.01.035"},{"key":"2026092312300928000_2026.09.12.751163v1.60","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2020.11.007"},{"key":"2026092312300928000_2026.09.12.751163v1.61","doi-asserted-by":"publisher","DOI":"10.1146\/annurev.immunol.19.1.497"},{"key":"2026092312300928000_2026.09.12.751163v1.62","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1406473111"},{"key":"2026092312300928000_2026.09.12.751163v1.63","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1413935112"},{"key":"2026092312300928000_2026.09.12.751163v1.64","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1402154"},{"key":"2026092312300928000_2026.09.12.751163v1.65","doi-asserted-by":"publisher","DOI":"10.1016\/j.celrep.2019.03.052"},{"key":"2026092312300928000_2026.09.12.751163v1.66","doi-asserted-by":"publisher","DOI":"10.1152\/physrev.00041.2020"},{"key":"2026092312300928000_2026.09.12.751163v1.67","doi-asserted-by":"publisher","DOI":"10.1096\/fj.201901136R"},{"key":"2026092312300928000_2026.09.12.751163v1.68","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-018-07416-2"},{"key":"2026092312300928000_2026.09.12.751163v1.69","doi-asserted-by":"publisher","DOI":"10.1126\/science.1241359"},{"key":"2026092312300928000_2026.09.12.751163v1.70","doi-asserted-by":"crossref","first-page":"987","DOI":"10.1016\/j.cmet.2020.04.007","article-title":"Developmental and Tumor Angiogenesis Requires the Mitochondria-Shaping Protein Opa1","volume":"31","year":"2020","journal-title":"Cell Metab"},{"key":"2026092312300928000_2026.09.12.751163v1.71","doi-asserted-by":"crossref","first-page":"1820","DOI":"10.1016\/j.cmet.2021.07.008","article-title":"Mitochondrial cristae-remodeling protein OPA1 in POMC neurons couples Ca(2+) homeostasis with adipose tissue lipolysis","volume":"33","year":"2021","journal-title":"Cell Metab"},{"key":"2026092312300928000_2026.09.12.751163v1.72","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2019.08.019"},{"key":"2026092312300928000_2026.09.12.751163v1.73","doi-asserted-by":"publisher","DOI":"10.1016\/j.cmet.2022.08.017"},{"key":"2026092312300928000_2026.09.12.751163v1.74","doi-asserted-by":"publisher","DOI":"10.1016\/j.molcel.2024.07.020"},{"key":"2026092312300928000_2026.09.12.751163v1.75","doi-asserted-by":"crossref","unstructured":"Pereira RO , Marti A , Olvera AC , Tadinada SM , Bjorkman SH , Weatherford ET , Morgan DA , Westphal M , Patel PH , Kirby AK , Hewezi R , Bui Tran W , Garcia-Pena LM , Souvenir RA , Mittal M , Adams CM , Rahmouni K , Potthoff MJ and Abel ED. OPA1 deletion in brown adipose tissue improves thermoregulation and systemic metabolism via FGF21. Elife. 2021;10.","DOI":"10.7554\/eLife.66519"},{"key":"2026092312300928000_2026.09.12.751163v1.76","doi-asserted-by":"crossref","first-page":"e14165","DOI":"10.1111\/acel.14165","article-title":"The integrated stress response promotes neural stem cell survival under conditions of mitochondrial dysfunction in neurodegeneration","volume":"23","year":"2024","journal-title":"Aging Cell"},{"key":"2026092312300928000_2026.09.12.751163v1.77","doi-asserted-by":"crossref","unstructured":"Dai W , Wang Z , Wang QA , Chan D and Jiang L . Metabolic reprogramming in the OPA1- deficient cells. Cell Mol Life Sci. 2022;79:517.","DOI":"10.1007\/s00018-022-04542-5"},{"key":"2026092312300928000_2026.09.12.751163v1.78","doi-asserted-by":"publisher","DOI":"10.1091\/mbc.E10-12-0933"},{"key":"2026092312300928000_2026.09.12.751163v1.79","doi-asserted-by":"publisher","DOI":"10.1083\/jcb.200903065"},{"key":"2026092312300928000_2026.09.12.751163v1.80","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.1801613"},{"key":"2026092312300928000_2026.09.12.751163v1.81","doi-asserted-by":"crossref","first-page":"941","DOI":"10.4049\/jimmunol.2400246","article-title":"Gpx4 Regulates Invariant NKT Cell Homeostasis and Function by Preventing Lipid Peroxidation and Ferroptosis","volume":"213","year":"2024","journal-title":"J Immunol"},{"key":"2026092312300928000_2026.09.12.751163v1.82","doi-asserted-by":"publisher","DOI":"10.1016\/j.jim.2009.07.009"}],"container-title":[],"original-title":[],"link":[{"URL":"https:\/\/syndication.highwire.org\/content\/doi\/10.64898\/2026.09.12.751163","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,23]],"date-time":"2026-09-23T19:30:22Z","timestamp":1790191822000},"score":1,"resource":{"primary":{"URL":"http:\/\/biorxiv.org\/lookup\/doi\/10.64898\/2026.09.12.751163"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,9,18]]},"references-count":82,"URL":"https:\/\/doi.org\/10.64898\/2026.09.12.751163","relation":{},"subject":[],"published":{"date-parts":[[2026,9,18]]},"subtype":"preprint"}}