Abstract Number: Esoc2026a2156 in Silico-Based Network Pharmacology Treatment Promotes Blood-Brain Barrier Stabilization and Reduced Neuroinflammation in a Murine Stroke Model

Abstract Background and aims Despite decades of extensive preclinical research, no neuroprotective therapy has successfully been translated into clinical practice to mitigate post-stroke disabilities. This translational gap could be attributed to a conservative, single-target, single-drug strategy insufficiently addressing the complexity of stroke pathophysiology. Systems medicine offers an alternative paradigm, suggesting that disease-associated proteins form interconnected modules within the human interactome. Accordingly, diseases should rather be defined as dysfunctional interacting network modules within the interactome, requiring simultaneous therapeutic targeting. Altogether, we here propose an in silico-based network pharmacology and drug repurposing approach to mechanistically target stroke-associated gene networks, aiming for a synergistic, supra-additive therapy while minimizing dosage and potential side effects. In this study, we specifically focus on blood-brain barrier integrity and neuroinflammatory response post-stroke. Methods Male C57BL/6N mice were subjected to 30 minutes of transient middle cerebral artery occlusion (tMCAO), followed by vehicle or the proposed network pharmacology treatment (NWPT). Tissue was later collected at 3, 7, or 14 days post-tMCAO. Blood-brain barrier integrity was assessed by analysing tight junction protein expression, while glial cell responses were evaluated for each timepoint. Results Upon treatment, the integrity of the blood-brain-barrier was improved with significantly increased expression of key tight junction proteins. Additionally, microglia activation and astrogliosis were markedly reduced in the treatment group during the sub-acute phase post-stroke. Conclusions The proposed in silico-based, multi-target disease module identification approach combined with a network pharmacology therapeutic strategy confers synergistic neuroprotection in the sub-chronic phase post-stroke, proposing network pharmacology as a promising framework to overcome the current translational therapeutic limitations. Conflict of interest Vonhof S.E.: Nothing to disclose