Multiple sclerosis (MS)–associated neurogenic bladder dysfunction is traditionally attributed to demyelination and loss of neural control. However, emerging data indicate that local bladder inflammation and immune–neural crosstalk actively contribute to disease progression. Building on previous work showing that inosine, a purine nucleoside with neuroprotective and anti-inflammatory properties, improved bladder function and reduced DNA damage following spinal cord injury (SCI) (1), we hypothesized that inosine may similarly attenuate inflammation-driven bladder dysfunction in experimental autoimmune encephalomyelitis (EAE) (2), the murine model of MS.

We collected bladders from EAE mice showing neurological symptoms and asymptomatic animals and assessed macrophage infiltration and other inflammatory changes by qPCR and shotgun proteomics. Using repeated urodynamic investigations (UDI) in awake mice we characterized bladder dysfunction in the EAE model of multiple sclerosis and correlated bladder function with symptom scores. To assess the anti-inflammatory potential of inosine, we performed luciferase assays in smooth muscle and urothelial cells stimulated with TNF-α in the presence or absence of inosine, followed by next-generation mRNA-seq (NGS) and comprehensive gene expression analysis. Finally, to evaluate the therapeutic effects of inosine in vivo, EAE mice were treated with inosine or vehicle, then surgically implanted with bladder catheters, and subjected to repeated UDI to monitor bladder function over time.

In the EAE model, bladder dysfunction was evidenced by an increased bladder-to-body weight ratio and abnormal urodynamic recordings. EAE bladders exhibited elevated expression of F4/80, CD68, and CCR2, consistent with pro-inflammatory macrophage infiltration and activation of IL-12–associated pathways. Proteomic analysis further demonstrated upregulation of inflammatory and fibrotic pathways, along with altered expression of proteins involved in macrophage activation (APOM, ITGAM), extracellular matrix remodeling (COL1A1–3, GATA3), and nitric oxide/reactive oxygen species production (ARG2, CYBB). In vitro, inosine significantly suppressed TNF-α–induced NF-κB activity in smooth muscle and urothelial cells. NGS analysis of stimulated cells revealed modulation of inflammatory pathways, with downregulation of key TNF signaling hubs including TIFA, DEPP1, and CEBPD. Additionally, inosine-treated smooth muscle cells showed upregulation of contractile markers ACTA1, FGF2, and EPHB3, suggesting preservation of muscle function. In vivo, inosine-treated EAE mice showed a reduced induction rate (10% vs. 40%) and decreased symptom severity, along with increased bladder capacity on repeated urodynamic investigations.

These findings indicate that EAE-associated bladder dysfunction is driven by inflammatory macrophage infiltration, activation of TNF/NF-κB–related signaling, and downstream fibrotic and oxidative stress pathways that collectively impair bladder structure and function. The upregulation of extracellular matrix remodeling and immune activation proteins supports the presence of chronic inflammation and tissue remodeling in the bladder. In contrast, inosine appears to exert a protective effect by attenuating NF-κB signaling, suppressing key components of the TNF inflammatory cascade, and preserving contractile phenotype in smooth muscle cells. The in vivo improvements in disease incidence, symptom severity, and bladder capacity further suggest that inosine mitigates both central disease processes and peripheral bladder dysfunction, highlighting its potential as a therapeutic agent targeting inflammation-driven neurogenic bladder impairment.

Our data reveal that EAE-induced bladder dysfunction arises from local inflammatory remodeling rather than neural degeneration alone. Consistent with our earlier findings in SCI, inosine suppresses inflammatory signaling, preserves smooth muscle phenotype, and improves bladder performance. These results identify inosine as a promising immunometabolic therapy for mitigating MS-associated lower urinary tract symptoms (LUTS) and potentially other neuroinflammatory bladder pathologies.

Multiomics analysis unveils an inosine-sensitive DNA damage response in neurogenic bladder after spinal cord injury. Ali Hashemi Gheinani, Bryan S. Sack, Alexander Bigger-Allen, Hatim Thaker,Hussein Atta, Rosalyn M. Adam. s.l. : JCI insight, 2025, Vol. 10.1172/jci.insight.180275.
Experimental autoimmune encephalomyelitis (EAE) as a model for multiple sclerosis (MS). Constantinescu CS, Farooqi N, O'Brien K, Gran B. s.l. : Br J Pharmacol., 2011. DOI: 10.1111/j.1476-5381.2011.01302.x.

Continence 19S (2026) 102698
DOI: 10.1016/j.cont.2026.102698