Li, Z.; Theocharidis, G.; Lee, S.; Kalavros, N.; Nagalakshmi, S. U.; Contreras, M.; Valsami, E.-A.; Nam, H.; Guan, M.; Chu, G.; Mezghani, I.; Wang, E.; Sumpio, B. J.; Hou, J.; Gennari-Felipe, M.; Gilman, J.; Lafuente-Gomez, N.; Maeso Aragon, L.; Dimitrakakis, N.; Arruda de Amaral, A.; Wang, S.; Vlachos, I. S.; Mooney, D. J.; Veves, A.

Chronic non-healing wounds are a major complication of diabetes, where persistent inflammation fails to transition into effective tissue repair (1-3). Here we identify cholinergic modulation as a mechanism that promotes productive inflammation and diabetic wound healing. Using a healing-associated fibroblast signature derived from single-cell profiling of human diabetic foot ulcers (3), we identified the acetylcholinesterase (AChE) inhibitor metrifonate as a candidate inducer of reparative wound programs. Topical delivery accelerated healing in diabetic mice and increased acetylcholine availability within the wound. Complementary receptor pharmacology identified muscarinic acetylcholine receptor 1 (CHRM1) as the principal receptor mediating the reparative response. Transcriptomic and cellular analyses revealed early induction of IL-17-associated inflammatory programs and enrichment of CHRM1NK1.1 innate lymphocytes. Depletion of NK1.1 cells or neutralization of IL-17A abolished the pro-healing effect, whereas spatial transcriptomics revealed treatment-associated organization of NK cells within vascular niches. Localized treatment further promoted repair in diabetic porcine wounds and human skin models. Together, these findings identify a CHRM1-IL-17 neuroimmune axis linking cholinergic modulation to diabetic wound repair and establish local AChE inhibition as a potential therapeutic strategy for chronic wounds.