Mesenchymal stem cell with enhanced antioxidant capacity integrates into smooth muscle cells in diabetic detrusor underactivity (Affymetrix)

Diabetic cystopathy, affecting half of individuals with diabetes, often progresses to detrusor underactivity (DUA), a late and irreversible stage of bladder dysfunction. Despite its prevalence, the etiology of diabetic DUA remains elusive, with no established treatments. Mesenchymal stem cell (MSC) transplantation, demonstrated as a promising therapeutic avenue in preclinical studies, lacks comprehensive analyses of its mechanisms, tumorigenic risks, and optimal protocols for diabetic DUA. This study addresses these gaps, exploring transcriptome changes post-MSC treatment in a diabetic DUA preclinical model. Oxidative injury emerged as a significant pathological mechanism, with N-acetylcysteine (NAC) enhancing MSC synergistic effects, potentially reducing the amount of required MSCs . Additionally, human umbilical cord-derived MSCs with high antioxidant capacity (PFO-MSCs) exhibited improved and sustained therapeutic efficacy, integrating into myocytes within bladder muscle bundles or neighboring pericytes. Single-cell transcriptome analysis of engrafted cells revealed expression changes in pathways related to muscle development and immune responses, emphasizing the pivotal roles of cMET and PD-L1 in muscle regeneration and immune modulation within diabetic DUA. This study provides a comprehensive overview of the pathogenesis and repair processes following MSC therapy for diabetic DUA, enhancing our understanding of its mode of action, efficacy, and safety for future clinical applications. Human umbilical cord-derived mesenchymal stem cells (UC-MSCs) were harvested in the normal cultured UC-MSCs (Cultured #) and the Resovist+/GFP+ engrafted cells (Engrafted #) from bladders of rats in which type I diabetes at 7 DAT. Total RNA from single UC-MSC engrafted rat bladder by MACS-FACS sorting was isolated using the cell lysis buffer, including treatment with NP40, Rnase Inhibitors, and V1(dT)24 primer(5 ng/ul). One ug of total RNA was used for the Affymetrix GeneChip Human Genome U133 Plus 2.0 Array (Affymetrix, Santa Clara, CA), in accordance with the manufacturer’s instructions.