{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,6]],"date-time":"2026-07-06T15:18:47Z","timestamp":1783351127834,"version":"3.54.6"},"reference-count":40,"publisher":"Wiley","issue":"6","license":[{"start":{"date-parts":[[2025,11,10]],"date-time":"2025-11-10T00:00:00Z","timestamp":1762732800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2025,11,10]],"date-time":"2025-11-10T00:00:00Z","timestamp":1762732800000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK067912"],"award-info":[{"award-number":["DK067912"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK1129712"],"award-info":[{"award-number":["DK1129712"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["DK102233"],"award-info":[{"award-number":["DK102233"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000002","name":"National Institutes of Health","doi-asserted-by":"publisher","award":["R01DK137515"],"award-info":[{"award-number":["R01DK137515"]}],"id":[{"id":"10.13039\/100000002","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000968","name":"American Heart Association","doi-asserted-by":"publisher","award":["25CDA1454374"],"award-info":[{"award-number":["25CDA1454374"]}],"id":[{"id":"10.13039\/100000968","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["J cachexia sarcopenia muscle"],"published-print":{"date-parts":[[2025,12]]},"abstract":"ABSTRACT<\/jats:title>\n \n Background<\/jats:title>\n Mitochondrial homeostasis is vital for optimal skeletal muscle integrity. Mitochondrial quality control (MQC) mechanisms that are essential for maintaining proper functions of mitochondria include mitochondrial biogenesis, dynamics and mitophagy. Previously, Syntaxin 4 (STX4), traditionally considered a cell surface protein known for glucose uptake in skeletal muscle, was also identified at the outer mitochondrial membrane. STX4 enrichment was sufficient to reverse Type 2 diabetes\u2013associated mitochondrial damage in skeletal muscle by inactivation of mitochondrial fission. However, whether STX4 could modulate skeletal muscle mitochondrial homeostasis through MQC mechanisms involving mitochondrial biogenesis or mitophagy remains to be determined.<\/jats:p>\n <\/jats:sec>\n \n Methods<\/jats:title>\n \n To determine the requirements of STX4 in mitochondrial structure, function and MQC processes of biogenesis and mitophagy, we implemented our in\u2010house generated inducible skeletal muscle\u2010specific STX4\u2010knockout (skmSTX4\u2010iKO) mice (\n Stx4<\/jats:italic>\n \n fl\/fl<\/jats:italic>\n <\/jats:sup>\n ; Tg (HSA\u2010rtTA\/TRE\u2010Cre<\/jats:italic>\n )\/B6) and STX4\u2010depleted immortalized L6.GLUT4myc myotubes via siRNA knockdown (siSTX4).\n <\/jats:p>\n <\/jats:sec>\n \n Results<\/jats:title>\n \n We found that non\u2010obese skmSTX4\u2010iKO male mice (>\u200950% reduced STX4 abundance, soleus and gastrocnemius ***\n p<\/jats:italic>\n \u2009<\u20090.001, tibialis anterior (TA) ****\n p<\/jats:italic>\n \u2009<\u20090.0001) developed insulin resistance (**\n p<\/jats:italic>\n \u2009<\u20090.01), together with reduced energy expenditure (AUC *\n p<\/jats:italic>\n \u2009<\u20090.05), respiratory exchange ratio (AUC **\n p<\/jats:italic>\n \u2009<\u20090.01) and grip strength (*\n p<\/jats:italic>\n \u2009<\u20090.05). STX4 ablation in muscle also impaired mitochondrial oxygen consumption rate (****\n p<\/jats:italic>\n \u2009<\u20090.0001). Mitochondrial morphological damage was heterogenous in STX4\u2010depleted muscle, presenting with small fragmented mitochondria (****\n p<\/jats:italic>\n \u2009<\u20090.0001) and decreased electron transport chain (ETC) abundance (CI ***\n p<\/jats:italic>\n \u2009<\u20090.001, CII *\n p<\/jats:italic>\n \u2009<\u20090.05, CIV **\n p<\/jats:italic>\n \u2009<\u20090.01) in oxidative soleus muscle, whereas glycolytic\u2010rich TA fibres displayed enlarged swollen mitochondria (****\n p<\/jats:italic>\n \u2009<\u20090.0001) with no change in ETC abundance. Notably, >\u200960% reduction of STX4 in siSTX4 L6.GLUT4myc myotubes (****\n p<\/jats:italic>\n \u2009<\u20090.0001) also decreased ETC abundance (CI **\n p<\/jats:italic>\n \u2009<\u20090.01, CII ***\n p<\/jats:italic>\n \u2009<\u20090.001, CIV **\n p<\/jats:italic>\n \u2009<\u20090.01) without changes in mitochondrial glucose metabolism, as shown by [U\u2010\n 13<\/jats:sup>\n C]glucose isotope tracing. For MQC, both skmSTX4\u2010iKO male mice (*\n p<\/jats:italic>\n \u2009<\u20090.05) and siSTX4 L6.GLUT4myc myotubes (*\n p<\/jats:italic>\n \u2009<\u20090.05) showed decreased mitochondrial DNA levels alongside reduced mRNA expression of mitochondrial biogenesis genes\n Ppargc1a<\/jats:italic>\n (PGC1\u2010\u03b1, *\n p<\/jats:italic>\n \u2009<\u20090.05) and\n Tfam<\/jats:italic>\n (*\n p<\/jats:italic>\n \u2009<\u20090.05) in skmSTX4\u2010iKO soleus muscle and PGC1\u2010\u03b1 (mRNA **\n p<\/jats:italic>\n \u2009<\u20090.01, protein *\n p<\/jats:italic>\n \u2009<\u20090.05), NRF1 (mRNA **\n p<\/jats:italic>\n < 0.01 and protein *\n p<\/jats:italic>\n \u2009<\u20090.05) and\n Tfam<\/jats:italic>\n (mRNA *\n p<\/jats:italic>\n \u2009<\u20090.05) in siSTX4 L6.GLUT4myc myotubes. Furthermore, live cell imaging using the mt\u2010Keima mitophagy biosensor in siSTX4 L6.GLUT4myc cells revealed significantly impaired mitochondrial turnover by mitophagy (*\n p<\/jats:italic>\n \u2009<\u20090.05) and mitochondria\u2013lysosome colocalization (*\n p<\/jats:italic>\n \u2009<\u20090.05). STX4 depletion also reduced canonical mitophagy markers, PINK1 and PARKIN in both skmSTX4\u2010iKO muscle (PARKIN *\n p<\/jats:italic>\n \u2009<\u20090.05, PINK1 **\n p<\/jats:italic>\n \u2009<\u20090.01) and siSTX4 L6.GLUT4myc myotubes (PARKIN **\n p<\/jats:italic>\n \u2009<\u20090.01, PINK1 *\n p<\/jats:italic>\n \u2009<\u20090.05).\n <\/jats:p>\n <\/jats:sec>\n \n Conclusions<\/jats:title>\n Our study demonstrated STX4 as a key mitochondrial regulator required for mitochondrial homeostasis in skeletal muscle.<\/jats:p>\n <\/jats:sec>","DOI":"10.1002\/jcsm.70113","type":"journal-article","created":{"date-parts":[[2025,11,11]],"date-time":"2025-11-11T05:26:32Z","timestamp":1762838792000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["STX4 Is Indispensable for Mitochondrial Homeostasis in Skeletal Muscle"],"prefix":"10.1002","volume":"16","author":[{"given":"Joseph\u00a0M.","family":"Hoolachan","sequence":"first","affiliation":[{"name":"Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute City of Hope Beckman Research Institute Duarte California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rekha","family":"Balakrishnan","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute City of Hope Beckman Research Institute Duarte California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Erika\u00a0M.","family":"McCown","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute City of Hope Beckman Research Institute Duarte California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karla\u00a0E.","family":"Merz","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute City of Hope Beckman Research Institute Duarte California USA"},{"name":"Amgen Inc Thousand Oaks California USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Chunxue","family":"Zhou","sequence":"additional","affiliation":[{"name":"Department of Molecular and Cellular Endocrinology, Arthur Riggs Diabetes and Metabolism Research Institute City of Hope Beckman Research Institute Duarte California USA"},{"name":"Sanegene Bio Inc. 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