Engineered Mesenchymal Stem Cells with Endogenous Trehalose Expression Activate the NRF2-HMOX1 Pathway to Enhance Antioxidant Stress and Wound Healing Capacity

Objective: Oxidative stress limits mesenchymal stem cell (MSC) efficacy in tissue repair by reducing retention and survival at injury sites. Endogenous production of trehalose may enhance MSC resilience and promote skin wound healing. Approach: Trehalose-6-phosphate synthase 1-expressing MSCs (TPS1-MSCs) were engineered via adenoviral transduction. Trehalose content and synthase activity were assessed. Oxidative stress models (H 2 O 2 , 0% fetal bovine serum, CoCl 2 ) were used to evaluate reactive oxygen species (ROS), apoptosis, and cell damage. TPS1-MSCs were transplanted into mouse wounds to track retention rate via in vivo imaging. Histology and immunofluorescence were used to assess wound healing, collagen deposition, and angiogenesis. Conditioned medium (CM) was used to evaluate paracrine functions. RNA-seq identified differentially expressed genes, and mechanisms were validated using the NRF2 inhibitor ML385. Results: TPS1-MSCs exhibited TPS1 activity and synthesized trehalose. Under oxidative stress, these cells showed reduced ROS, enhanced viability, and decreased apoptosis. In vivo , TPS1-MSCs displayed higher retention, accelerated healing, and neovascularization. CM from TPS1-MSCs promoted keratinocyte migration, fibroblast collagen secretion, and enhanced the tube-forming capacity of endothelial cells. Transcriptome analysis revealed enrichment in the NRF2-HMOX1 pathway. TPS1-MSCs showed elevated levels of p62, nuclear NRF2, and HMOX1. ML385 treatment impaired the observed antioxidant capacity. Innovation: Engineering MSCs for endogenous trehalose synthesis enhanced oxidative stress resistance and retention through NRF2-HMOX1 activation, suggesting a potential novel MSC-based wound repair strategy. Conclusion: TPS1-MSCs improved antioxidant capacity and wound healing, potentially through the NRF2-HMOX1 pathway, and may represent a promising therapy for skin wounds. Guangchao Xu, PhD Rongqing Pang, PhD