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Our previous study showed that mitochondrial fission protein dynamin-related protein 1 (DRP1) promotes reparative polarization and metabolic reprogramming in macrophages and post-ischemic neovascularization. However, the redox-dependent mechanism governing DRP1 activation in this context remains elusive. Here, using a mouse hindlimb ischemia (HLI) model of PAD, we identify cysteine sulfenylation (CysOH) of DRP1 as a critical redox modification induced in ischemic bone marrow (BM)-derived cells. BM chimeric mice reconstituted with CRISPR\/Cas9-generated \u201credox-dead\u201d DRP1-C631A knock-in mutant (Drp1C\/A) BM exhibited markedly reduced limb perfusion recovery and CD31+ capillary density in ischemic muscles following HLI. These defects were associated with enhanced Ly6G+ neutrophil accumulation, pro-inflammatory F4\/80+CD80+ M1-like macrophages and reduced anti-inflammatory F4\/80+CD206+ M2-like macrophages in ischemic muscle. Mechanistically, using an in vitro PAD model, hypoxia serum starvation (HSS) rapidly induced NADPH oxidase 2-dependent cytosolic ROS production and DRP1-CysOH formation in wild-type macrophages. In contrast, Drp1C\/A macrophages failed to undergo DRP1-CysOH-dependent mitochondrial fission under HSS, resulting in aberrant metabolic reprogramming characterized by enhanced glycolysis and mitochondrial ROS, pro-inflammatory p-NF-\u03baB and M1-genes, and suppressed anti-inflammatory p-AMPK, efferocytosis and M2-genes. Thus, our findings establish DRP1 sulfenylation as a previously unrecognized redox-sensing mechanism that links ischemia-induced ROS to reparative macrophage reprogramming and revascularization, identifying a novel therapeutic target for PAD.<\/jats:p>","DOI":"10.3390\/antiox15060768","type":"journal-article","created":{"date-parts":[[2026,6,26]],"date-time":"2026-06-26T05:52:11Z","timestamp":1782453131000},"page":"768","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Myeloid DRP1 Sulfenylation Drives Reparative Macrophage Polarization and Neovascularization in Ischemic Muscle"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-8195-5700","authenticated-orcid":false,"given":"Shikha","family":"Yadav","sequence":"first","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajagopal","family":"Kamarajan","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9675-0097","authenticated-orcid":false,"given":"Varadarajan","family":"Sudhahar","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Charlie Norwood Veterans Affairs Medical Center, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3707-5036","authenticated-orcid":false,"given":"Sheela","family":"Nagarkoti","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Archita","family":"Das","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Charlie Norwood Veterans Affairs Medical Center, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0003-3355-3111","authenticated-orcid":false,"given":"Stephanie Kelley","family":"Spears","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajalakshmi Veeranan","family":"Karmegam","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-9864-571X","authenticated-orcid":false,"given":"Tohru","family":"Fukai","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Charlie Norwood Veterans Affairs Medical Center, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7048-2381","authenticated-orcid":false,"given":"Masuko","family":"Ushio-Fukai","sequence":"additional","affiliation":[{"name":"Vascular Biology Center, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"},{"name":"Department of Medicine (Cardiology), Medical College of Georgia, Augusta University, Augusta, GA 30912, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,6,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"e1020","DOI":"10.1016\/S2214-109X(19)30255-4","article-title":"Global, regional, and national prevalence and risk factors for peripheral artery disease in 2015: An updated systematic review and analysis","volume":"7","author":"Song","year":"2019","journal-title":"Lancet Glob. 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