{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,9]],"date-time":"2026-03-09T14:36:20Z","timestamp":1773066980198,"version":"3.50.1"},"reference-count":43,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2021,12,31]],"date-time":"2021-12-31T00:00:00Z","timestamp":1640908800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Both Type 1 diabetes mellitus (DM1) and type 2 diabetes mellitus (DM2) are associated with an increased risk of limb amputation in peripheral arterial disease (PAD). How diabetes contributes to poor PAD outcomes is poorly understood but may occur through different mechanisms in DM1 and DM2. Previously, we identified a disintegrin and metalloproteinase gene 12 (ADAM12) as a key genetic modifier of post-ischemic perfusion recovery. In an experimental PAD, we showed that ADAM12 is regulated by miR-29a and this regulation is impaired in ischemic endothelial cells in DM1, contributing to poor perfusion recovery. Here we investigated whether miR-29a regulation of ADAM12 is altered in experimental PAD in the setting of DM2. We also explored whether modulation of miR-29a and ADAM12 expression can improve perfusion recovery and limb function in mice with DM2. Our result showed that in the ischemic limb of mice with DM2, miR-29a expression is poorly downregulated and ADAM12 upregulation is impaired. Inhibition of miR-29a and overexpression of ADAM12 improved perfusion recovery, reduced skeletal muscle injury, improved muscle function, and increased cleaved Tie 2 and AKT phosphorylation. Thus, inhibition of miR-29a and or augmentation of ADAM12 improves experimental PAD outcomes in DM2 likely through modulation of Tie 2 and AKT signalling.<\/jats:p>","DOI":"10.3390\/ijms23010429","type":"journal-article","created":{"date-parts":[[2022,1,3]],"date-time":"2022-01-03T20:49:38Z","timestamp":1641242978000},"page":"429","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":17,"title":["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"],"prefix":"10.3390","volume":"23","author":[{"given":"Victor","family":"Lamin","sequence":"first","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Joseph","family":"Verry","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Isaac","family":"Eigner-Bybee","sequence":"additional","affiliation":[{"name":"Division of Endocrinology and Metabolism, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4437-0834","authenticated-orcid":false,"given":"Jordan D.","family":"Fuqua","sequence":"additional","affiliation":[{"name":"Department of Health and Human Physiology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"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":[{"role":"author","vocabulary":"crossref"}]},{"given":"Vitor A.","family":"Lira","sequence":"additional","affiliation":[{"name":"Department of Health and Human Physiology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IA 52242, USA"},{"name":"Fraternal Order of Eagles Diabetes Research Center, Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"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":"Department of Health and Human Physiology, College of Liberal Arts and Sciences, University of Iowa, Iowa City, IA 52242, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,12,31]]},"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":"1608","DOI":"10.1056\/NEJM200105243442108","article-title":"Medical treatment of peripheral arterial disease and claudication","volume":"344","author":"Hiatt","year":"2001","journal-title":"N. Engl. J. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1944","DOI":"10.1056\/NEJMe068037","article-title":"Treatment of peripheral arterial disease--extending \u201cintervention\u201d to \u201ctherapeutic choice\u201d","volume":"354","author":"Hirsch","year":"2006","journal-title":"N. Engl. J. Med."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"S5","DOI":"10.1016\/j.jvs.2006.12.037","article-title":"Inter-Society Consensus for the Management of Peripheral Arterial Disease (TASC II)","volume":"45","author":"Norgren","year":"2007","journal-title":"J. Vasc. Surg."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1038\/nrcardio.2013.70","article-title":"Therapeutic angiogenesis for critical limb ischaemia","volume":"10","author":"Annex","year":"2013","journal-title":"Nat. Rev. Cardiol."},{"key":"ref_16","first-page":"224","article-title":"Towards the Development of Novel Therapeutics for Peripheral Artery Disease","volume":"127","author":"Annex","year":"2016","journal-title":"Trans. Am. Clin. Climatol. Assoc."},{"key":"ref_17","first-page":"A141","article-title":"The incidence and health economic burden of ischemic amputation in Minnesota, 2005\u20132008","volume":"8","author":"Peacock","year":"2011","journal-title":"Prev. Chronic. Dis."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2241","DOI":"10.1161\/CIRCULATIONAHA.113.000502","article-title":"Approach to the patient with peripheral arterial disease","volume":"128","author":"Wennberg","year":"2013","journal-title":"Circulation"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1089\/hum.2006.17.683","article-title":"Treatment with intramuscular vascular endothelial growth factor gene compared with placebo for patients with diabetes mellitus and critical limb ischemia: A double-blind randomized trial","volume":"17","author":"Kusumanto","year":"2006","journal-title":"Hum. Gene Ther."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1006\/mthe.2002.0638","article-title":"Increased vascularity detected by digital subtraction angiography after VEGF gene transfer to human lower limb artery: A randomized, placebo-controlled, double-blinded phase II study","volume":"6","author":"Makinen","year":"2002","journal-title":"Mol. Ther."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"392","DOI":"10.1038\/gt.2011.109","article-title":"10-year safety follow-up in patients with local VEGF gene transfer to ischemic lower limb","volume":"19","author":"Muona","year":"2012","journal-title":"Gene Ther."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"972","DOI":"10.1038\/mt.2008.33","article-title":"Therapeutic Angiogenesis With Intramuscular NV1FGF Improves Amputation-free Survival in Patients With Critical Limb Ischemia","volume":"16","author":"Nikol","year":"2008","journal-title":"Mol. Ther."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1933","DOI":"10.1161\/01.CIR.0000093398.16124.29","article-title":"Regional angiogenesis with vascular endothelial growth factor in peripheral arterial disease: A phase II randomized, double-blind, controlled study of adenoviral delivery of vascular endothelial growth factor 121 in patients with disabling intermittent claudication","volume":"108","author":"Rajagopalan","year":"2003","journal-title":"Circulation"},{"key":"ref_24","first-page":"970607","article-title":"Mechanisms of miRNA-Mediated Gene Regulation from Common Downregulation to mRNA-Specific Upregulation","volume":"2014","author":"Safaralizadeh","year":"2014","journal-title":"Int. J. Genom."},{"key":"ref_25","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_26","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_27","doi-asserted-by":"crossref","first-page":"948","DOI":"10.1161\/CIRCRESAHA.107.160630","article-title":"Impaired angiogenesis after hindlimb ischemia in type 2 diabetes mellitus: Differential regulation of vascular endothelial growth factor receptor 1 and soluble vascular endothelial growth factor receptor 1","volume":"101","author":"Hazarika","year":"2007","journal-title":"Circ. Res."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"656","DOI":"10.2337\/db06-0999","article-title":"In mice with type 2 diabetes, a vascular endothelial growth factor (VEGF)-activating transcription factor modulates VEGF signaling and induces therapeutic angiogenesis after hindlimb ischemia","volume":"56","author":"Li","year":"2007","journal-title":"Diabetes"},{"key":"ref_29","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."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3422","DOI":"10.1073\/pnas.97.7.3422","article-title":"Transplantation of ex vivo expanded endothelial progenitor cells for therapeutic neovascularization","volume":"97","author":"Kalka","year":"2000","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1408","DOI":"10.1089\/scd.2012.0445","article-title":"Endothelial progenitor cells derived from Wharton\u2019s jelly of the umbilical cord reduces ischemia-induced hind limb injury in diabetic mice by inducing HIF-1alpha\/IL-8 expression","volume":"22","author":"Shen","year":"2013","journal-title":"Stem. Cells Dev."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"137","DOI":"10.1186\/1479-5876-10-137","article-title":"Obesity suppresses circulating level and function of endothelial progenitor cells and heart function","volume":"10","author":"Tsai","year":"2012","journal-title":"J. Transl. Med."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Ieronimakis, N., Balasundaram, G., and Reyes, M. (2008). Direct isolation, culture and transplant of mouse skeletal muscle derived endothelial cells with angiogenic potential. PLoS ONE, 3.","DOI":"10.1371\/journal.pone.0001753"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1042\/BJ20121558","article-title":"ADAM12 is expressed in the tumour vasculature and mediates ectodomain shedding of several membrane-anchored endothelial proteins","volume":"452","author":"Frohlich","year":"2013","journal-title":"Biochem. J."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Lekas, M., Lekas, P., Mei, S.H., Deng, Y., Dumont, D.J., and Stewart, D.J. (2012). Tie2-dependent neovascularization of the ischemic hindlimb is mediated by angiopoietin-2. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0043568"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2619","DOI":"10.1161\/ATVBAHA.107.150482","article-title":"VEGF induces Tie2 shedding via a phosphoinositide 3-kinase\/Akt dependent pathway to modulate Tie2 signaling","volume":"27","author":"Findley","year":"2007","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1415","DOI":"10.1182\/blood-2010-03-275289","article-title":"Epithin\/PRSS14 proteolytically regulates angiopoietin receptor Tie2 during transendothelial migration","volume":"117","author":"Kim","year":"2011","journal-title":"Blood"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"215","DOI":"10.1016\/j.cell.2009.01.002","article-title":"MicroRNAs: Target recognition and regulatory functions","volume":"136","author":"Bartel","year":"2009","journal-title":"Cell"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"160","DOI":"10.18632\/aging.100643","article-title":"MicroRNA-29 induces cellular senescence in aging muscle through multiple signaling pathways","volume":"6","author":"Hu","year":"2014","journal-title":"Aging"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"768","DOI":"10.1002\/stem.2281","article-title":"MicroRNA-29a in Adult Muscle Stem Cells Controls Skeletal Muscle Regeneration During Injury and Exercise Downstream of Fibroblast Growth Factor-2","volume":"34","author":"Galimov","year":"2016","journal-title":"Stem. Cells"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"367","DOI":"10.1089\/hum.2019.287","article-title":"Exogenous miR-29a Attenuates Muscle Atrophy and Kidney Fibrosis in Unilateral Ureteral Obstruction Mice","volume":"31","author":"Wang","year":"2020","journal-title":"Hum. Gene Ther."},{"key":"ref_42","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_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."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/1\/429\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T07:56:39Z","timestamp":1760169399000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/23\/1\/429"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,12,31]]},"references-count":43,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,1]]}},"alternative-id":["ijms23010429"],"URL":"https:\/\/doi.org\/10.3390\/ijms23010429","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,12,31]]}}}