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bioRxiv · 10.64898/2026.02.15.706043

Myeloid DRP1 Sulfenylation Drives Reparative Macrophage Polarization and Neovascularization in Ischemic Muscle

Yadav, S.·Nagarkoti, S.·Sudhahar, V.·Rajagopal, K.·Das, A.·Spears, S. K.·Fukai, T.·Ushio-Fukai, M.
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Abstract

Reparative macrophage polarization and macrophage-derived reactive oxygen species (ROS) are required for ischemia-induced revascularization in peripheral artery disease (PAD). Our previous study showed that mitochondrial fission protein 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 "redox-dead" 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 F4/80CD80 M1 macrophages and reduced anti-inflammatory F4/80CD206 M2 macrophages in ischemic muscle. Mechanistically, using an in vitro PAD model, hypoxia-serum starvation (HSS) rapidly induced 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-{kappa}B and M1-genes, and suppressed anti-inflammatory p-AMPK 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.

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Yadav, S., Nagarkoti, S., Sudhahar, V., Rajagopal, K., Das, A., Spears, S. K., Fukai, T., Ushio-Fukai, M.. 2026-02-17. Myeloid DRP1 Sulfenylation Drives Reparative Macrophage Polarization and Neovascularization in Ischemic Muscle. https://doi.org/10.64898/2026.02.15.706043

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