CD73-Overexpressing ADSCs Promote Neurogenic Bladder Repair via A2A-JAK1/STAT3-Mediated Modulation of Inflammation and Macrophage Polarization.

Abstract

Background: Neurogenic bladder (NB) is characterized by chronic inflammation and bladder tissue injury, and current treatments fail to restore the inflammatory microenvironment or tissue integrity. Although adipose-derived stem cell (ADSC) therapy shows regenerative potential, its efficacy is limited by inflammation; whether CD73 overexpression enhances ADSC therapeutic effects in NB remains unclear.

Methods: A rat model of NB was established via pelvic nerve (PN) injury and treated with CD73-overexpressing ADSCs (CD73⁺ADSCs). Bladder function, histological remodeling, and apoptosis were evaluated in vivo. In vitro, conditioned medium (CM) from CD73⁺ADSCs was applied to LPS-injured rat bladder transitional epithelial cells (RB-TECs) and macrophages to assess proliferation, migration, apoptosis, inflammatory responses, and macrophage polarization. Key signaling pathways were examined, and label-free quantitative (LFQ) proteomics was performed to characterize global molecular reprogramming.

Results: CD73⁺ADSCs significantly improved urodynamic function, restored smooth muscle architecture, and reduced apoptosis in NB rats. In vitro, CD73⁺ADSC-CM promoted urothelial cell proliferation and migration while attenuating cell death. Mechanistically, CD73⁺ADSCs enhanced extracellular adenosine production, at least partly dependent on pharmacologically sensitive A2A receptor signaling, suppressed NF-κB activation, reduced proinflammatory cytokine release, promoted macrophage polarization toward an M2 phenotype, and inhibited NLRP3 inflammasome activation. Proteomic analysis revealed a global shift toward antiinflammatory and reparative molecular programs.

Conclusion: Our findings suggest that CD73 contributes to the immunomodulatory and reparative properties of ADSCs under inflammatory conditions. By coordinating adenosinergic signaling, inflammatory suppression, and repair-associated pathways, CD73⁺ADSCs effectively reprogram the NB microenvironment and represent a promising regenerative therapeutic strategy for NB.