{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T21:05:16Z","timestamp":1783458316951,"version":"3.55.0"},"reference-count":45,"publisher":"MDPI AG","issue":"18","license":[{"start":{"date-parts":[[2022,9,14]],"date-time":"2022-09-14T00:00:00Z","timestamp":1663113600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"National Heart, Lung, and Blood Institute","award":["R01 HL130399"],"award-info":[{"award-number":["R01 HL130399"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Peripheral artery disease (PAD) is characterized by impaired blood flow to the lower extremities, resulting in ischemic limb injuries. Individuals with diabetes and PAD typically have more severe ischemic limb injuries and limb amputations, but the mechanisms involved are poorly understood. Previously, we identified BAG3 as a gene within a mouse genetic locus termed limb salvage QTL1 on mouse chromosome 7 that determined the extent of limb necrosis following ischemic injury in C57Bl\/6 mice. Whether BAG3 deficiency plays a role in the severe ischemic injury observed in diabetic PAD is not known. In vitro, we found simulated ischemia enhanced BAG3 expression in primary human skeletal muscle cells, whereas BAG3 knockdown increased necroptosis markers and decreased cell viability. In vivo, ischemic skeletal muscles from hind limbs of high-fat diet (HFD)-fed mice showed poor BAG3 expression compared to normal chow diet (NCD)-fed mice, and this was associated with increased limb amputations. BAG3 overexpression in ischemic skeletal muscles from hind limbs of HFD mice rescued limb amputation and improved autophagy, necroptosis, skeletal muscle function and regeneration. Therefore, BAG3 deficiency in ischemic skeletal muscles contributes to the severity of ischemic limb injury in diabetic PAD, likely through autophagy and necroptosis pathways.<\/jats:p>","DOI":"10.3390\/ijms231810715","type":"journal-article","created":{"date-parts":[[2022,9,14]],"date-time":"2022-09-14T23:16:36Z","timestamp":1663197396000},"page":"10715","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["BAG3 Attenuates Ischemia-Induced Skeletal Muscle Necroptosis in Diabetic Experimental Peripheral Artery Disease"],"prefix":"10.3390","volume":"23","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8943-7406","authenticated-orcid":false,"given":"Arul M.","family":"Mani","sequence":"first","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karthik","family":"Dhanabalan","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Victor","family":"Lamin","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3234-8522","authenticated-orcid":false,"given":"Thomas","family":"Wong","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3048-4432","authenticated-orcid":false,"given":"Madhu V.","family":"Singh","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ayotunde O.","family":"Dokun","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"},{"name":"Fraternal Order of Eagles Diabetes Research Centre, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,9,14]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1055\/s-0031-1278244","article-title":"Peripheral arterial disease: Epidemiology, natural history, diagnosis and treatment","volume":"16","author":"Dhaliwal","year":"2007","journal-title":"Int. J. Angiol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1329","DOI":"10.1016\/S0140-6736(13)61249-0","article-title":"Comparison of global estimates of prevalence and risk factors for peripheral artery disease in 2000 and 2010: A systematic review and analysis","volume":"382","author":"Fowkes","year":"2013","journal-title":"Lancet"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1312","DOI":"10.1016\/S0140-6736(13)61576-7","article-title":"The global pandemic of peripheral artery disease","volume":"382","author":"Hirsch","year":"2013","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1111\/j.1464-5491.2009.02866.x","article-title":"Peripheral arterial disease in diabetes\u2014A review","volume":"27","author":"Jude","year":"2010","journal-title":"Diabet. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2436","DOI":"10.1093\/eurheartj\/eht149","article-title":"Diabetes and vascular disease: Pathophysiology, clinical consequences, and medical therapy: Part I","volume":"34","author":"Paneni","year":"2013","journal-title":"Eur. Heart J."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"961","DOI":"10.4239\/wjd.v6.i7.961","article-title":"Peripheral artery disease in patients with diabetes: Epidemiology, mechanisms, and outcomes","volume":"6","author":"Thiruvoipati","year":"2015","journal-title":"World J. Diabetes"},{"key":"ref_7","first-page":"200","article-title":"Impact of atherosclerotic risk factors on the anatomical distribution of peripheral arterial disease","volume":"20","author":"Haltmayer","year":"2001","journal-title":"Int. Angiol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1433","DOI":"10.2337\/diacare.24.8.1433","article-title":"Peripheral arterial disease in diabetic and nondiabetic patients: A comparison of severity and outcome","volume":"24","author":"Jude","year":"2001","journal-title":"Diabetes Care"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"921","DOI":"10.1016\/j.jacc.2005.09.065","article-title":"Peripheral arterial disease in patients with diabetes","volume":"47","author":"Marso","year":"2006","journal-title":"J. Am. Coll. Cardiol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"650","DOI":"10.2337\/diacare.3.6.650","article-title":"Incidence and prevalence of clinical peripheral vascular disease in a population-based cohort of diabetic patients","volume":"3","author":"Melton","year":"1980","journal-title":"Diabetes Care"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"364","DOI":"10.1093\/cvr\/cvt342","article-title":"Glycaemic control improves perfusion recovery and VEGFR2 protein expression in diabetic mice following experimental PAD","volume":"101","author":"Dokun","year":"2014","journal-title":"Cardiovasc. Res."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1432","DOI":"10.1177\/1535370217716424","article-title":"Modulation of miR29a improves impaired post-ischemic angiogenesis in hyperglycemia","volume":"242","author":"Chen","year":"2017","journal-title":"Exp. Biol. Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Lamin, V., Verry, J., Eigner-Bybee, I., Fuqua, J.D., Wong, T., Lira, V.A., and Dokun, A.O. (2021). Modulation of miR-29a and ADAM12 Reduces Post-Ischemic Skeletal Muscle Injury and Improves Perfusion Recovery and Skeletal Muscle Function in a Mouse Model of Type 2 Diabetes and Peripheral Artery Disease. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23010429"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1161\/01.CIR.99.6.807","article-title":"Skeletal muscle mitochondrial DNA injury in patients with unilateral peripheral arterial disease","volume":"99","author":"Bhat","year":"1999","journal-title":"Circulation"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1177\/1358836X0000500109","article-title":"Acquired skeletal muscle metabolic myopathy in atherosclerotic peripheral arterial disease","volume":"5","author":"Brass","year":"2000","journal-title":"Vasc. Med."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"58","DOI":"10.1161\/01.RES.0000133247.69803.c3","article-title":"Vascular endothelial growth factor overexpression in ischemic skeletal muscle enhances myoglobin expression in vivo","volume":"95","author":"Deckers","year":"2004","journal-title":"Circ. Res."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1393","DOI":"10.1016\/S0002-9440(10)62566-7","article-title":"Expression of vascular endothelial growth factor and vascular endothelial growth factor receptor-2 (KDR\/Flk-1) in ischemic skeletal muscle and its regeneration","volume":"160","author":"Rissanen","year":"2002","journal-title":"Am. J. Pathol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2156","DOI":"10.1016\/j.ajpath.2012.01.032","article-title":"Skeletal muscle-specific genetic determinants contribute to the differential strain-dependent effects of hindlimb ischemia in mice","volume":"180","author":"McClung","year":"2012","journal-title":"Am. J. Pathol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"336","DOI":"10.3389\/fphys.2015.00336","article-title":"Mitochondrial Regulation of the Muscle Microenvironment in Critical Limb Ischemia","volume":"6","author":"Ryan","year":"2015","journal-title":"Front. Physiol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"H790","DOI":"10.1152\/ajpheart.00803.2014","article-title":"ADAM12: A genetic modifier of preclinical peripheral arterial disease","volume":"309","author":"Dokun","year":"2015","journal-title":"Am. J. Physiol. Heart Circ. Physiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1207","DOI":"10.1161\/CIRCULATIONAHA.107.736447","article-title":"A quantitative trait locus (LSq-1) on mouse chromosome 7 is linked to the absence of tissue loss after surgical Hindlimb ischemia","volume":"117","author":"Md","year":"2008","journal-title":"Circulation"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"281","DOI":"10.1161\/CIRCULATIONAHA.116.024873","article-title":"BAG3 (Bcl-2-Associated Athanogene-3) Coding Variant in Mice Determines Susceptibility to Ischemic Limb Muscle Myopathy by Directing Autophagy","volume":"136","author":"McClung","year":"2017","journal-title":"Circulation"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"190","DOI":"10.1177\/1535370217743460","article-title":"Role of genetics in peripheral arterial disease outcomes; significance of limb-salvage quantitative locus-1 genes","volume":"243","author":"Okeke","year":"2018","journal-title":"Exp. Biol. Med."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"e149415","DOI":"10.1172\/JCI149415","article-title":"Therapeutic targeting of BAG3: Considering its complexity in cancer and heart disease","volume":"131","author":"Kirk","year":"2021","journal-title":"J. Clin. Investig."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"761","DOI":"10.2353\/ajpath.2006.060250","article-title":"BAG3 deficiency results in fulminant myopathy and early lethality","volume":"169","author":"Homma","year":"2006","journal-title":"Am. J. Pathol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1002\/ana.21553","article-title":"Mutation in BAG3 causes severe dominant childhood muscular dystrophy","volume":"65","author":"Selcen","year":"2009","journal-title":"Ann. Neurol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1074\/jbc.M706304200","article-title":"HspB8 chaperone activity toward poly(Q)-containing proteins depends on its association with Bag3, a stimulator of macroautophagy","volume":"283","author":"Carra","year":"2008","journal-title":"J. Biol. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"237","DOI":"10.4161\/auto.5407","article-title":"HspB8 and Bag3: A new chaperone complex targeting misfolded proteins to macroautophagy","volume":"4","author":"Carra","year":"2008","journal-title":"Autophagy"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1042\/BJ20090907","article-title":"Identification of the key structural motifs involved in HspB8\/HspB6-Bag3 interaction","volume":"425","author":"Fuchs","year":"2009","journal-title":"Biochem. J."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1007\/s00109-011-0795-6","article-title":"Emerging roles of molecular chaperones and co-chaperones in selective autophagy: Focus on BAG proteins","volume":"89","author":"Gamerdinger","year":"2011","journal-title":"J. Mol. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"889","DOI":"10.1038\/emboj.2009.29","article-title":"Protein quality control during aging involves recruitment of the macroautophagy pathway by BAG3","volume":"28","author":"Gamerdinger","year":"2009","journal-title":"EMBO J."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1038\/cdd.2015.171","article-title":"Cell death, clearance and immunity in the skeletal muscle","volume":"23","author":"Sciorati","year":"2016","journal-title":"Cell Death Differ."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1650","DOI":"10.1159\/000489241","article-title":"Current Mechanistic Concepts in Ischemia and Reperfusion Injury","volume":"46","author":"Wu","year":"2018","journal-title":"Cell. Physiol. Biochem."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"3655","DOI":"10.1038\/s41467-018-06057-9","article-title":"Necroptosis mediates myofibre death in dystrophin-deficient mice","volume":"9","author":"Morgan","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"e44452","DOI":"10.7554\/eLife.44452","article-title":"Autophagy regulates inflammatory programmed cell death via turnover of RHIM-domain proteins","volume":"8","author":"Lim","year":"2019","journal-title":"eLife"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1186\/s12933-021-01441-0","article-title":"Diabetes and other vascular risk factors in association with the risk of lower extremity amputation in chronic limb-threatening ischemia: A prospective cohort study","volume":"21","author":"Ying","year":"2022","journal-title":"Cardiovasc. Diabetol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1808","DOI":"10.1161\/ATVBAHA.120.314595","article-title":"Epidemiology and Risk of Amputation in Patients with Diabetes Mellitus and Peripheral Artery Disease","volume":"40","author":"Barnes","year":"2020","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"e141","DOI":"10.1038\/cddis.2011.24","article-title":"BAG3: A multifaceted protein that regulates major cell pathways","volume":"2","author":"Rosati","year":"2011","journal-title":"Cell Death Dis."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5153","DOI":"10.1038\/onc.2012.17","article-title":"BAG3 controls angiogenesis through regulation of ERK phosphorylation","volume":"31","author":"Falco","year":"2012","journal-title":"Oncogene"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"491","DOI":"10.1159\/000445641","article-title":"Overexpression of BAG3 Attenuates Hypoxia-Induced Cardiomyocyte Apoptosis by Inducing Autophagy","volume":"39","author":"Zhang","year":"2016","journal-title":"Cell Physiol. Biochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"516518","DOI":"10.1155\/2014\/516518","article-title":"BAG3 and HIF-1 alpha coexpression detected by immunohistochemistry correlated with prognosis in hepatocellular carcinoma after liver transplantation","volume":"2014","author":"Xiao","year":"2014","journal-title":"BioMed Res. Int."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1177\/1358863X19866254","article-title":"Dual specificity phosphatase 5 regulates perfusion recovery in experimental peripheral artery disease","volume":"24","author":"Alleboina","year":"2019","journal-title":"Vasc. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1016\/j.ab.2013.05.027","article-title":"Stain-Free total protein staining is a superior loading control to beta-actin for Western blots","volume":"440","author":"Gilda","year":"2013","journal-title":"Anal. Biochem."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"11735","DOI":"10.1096\/fj.201900766R","article-title":"ULK2 is essential for degradation of ubiquitinated protein aggregates and homeostasis in skeletal muscle","volume":"33","author":"Fuqua","year":"2019","journal-title":"FASEB J."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"363","DOI":"10.3389\/fphys.2013.00363","article-title":"Nuclear positioning in muscle development and disease","volume":"4","author":"Folker","year":"2013","journal-title":"Front. Physiol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/18\/10715\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T00:31:18Z","timestamp":1760142678000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/18\/10715"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,9,14]]},"references-count":45,"journal-issue":{"issue":"18","published-online":{"date-parts":[[2022,9]]}},"alternative-id":["ijms231810715"],"URL":"https:\/\/doi.org\/10.3390\/ijms231810715","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,9,14]]}}}