Multi‐Omics Integration Reveals That SN‐011 Targets JUNB to Upregulate ADGRE5 and Restore Vascular Endothelial Cell Communication in Ischemic Stroke

ABSTRACT

Ischemic stroke (IS) is a cerebrovascular disease with high mortality and disability rates, currently lacking effective therapeutic targets. The STING inhibitor SN‐011 shows potential in IS treatment, but its mechanism of action remains unclear. This study aims to explore the key molecular mechanisms of SN‐011 in treating IS through bioinformatics approaches. IS transcriptome datasets were analyzed to identify differentially expressed genes. Mendelian randomization using brain eQTL and IS‐GWAS data identified genes with causal relationships to IS. Single‐cell transcriptome, pseudo‐time trajectory, intercellular communication, and transcription factor regulatory network analyzes were performed. Molecular docking and DARTS‐WB assay validated SN‐011 binding to transcription factors. Transcriptomic analysis identified 77 intersecting genes. Mendelian randomization revealed ADGRE5 as a protective gene for IS (OR < 1), significantly downregulated in venous endothelial cells (vECs) during disease progression. Cell communication analysis showed ADGRE5‐high vECs interact with immune, glial, and stromal cells via LAMININ (Lamb2‐CD44, Lamb2‐Itga6+Itgb1, Lamb2‐Dag1 pair) and JAM signaling pathways. Transcription factor analysis identified JUNB as a negative regulator of ADGRE5. Molecular docking (−6.8 kcal/mol) combined with an in vitro DARTS‐WB assay confirmed the interaction between SN‐011 and JUNB. In OGD‐induced endothelial cell injury models, SN‐011 suppressed JUNB expression, restored ADGRE5 expression inhibited by JUNB overexpression, reversed the downregulation of LAMB2 and CD44, and reduced the expression of the pro‐inflammatory cytokines IL‐6 and IL‐1β. Notably, blockade of LAMB2 largely abolished these protective effects, indicating that the anti‐inflammatory and endothelial‐protective activities of SN‐011 are mediated, at least in part, through restoration of the LAMB2‐CD44 signaling axis. ADGRE5 downregulation in vECs may impair vascular repair by disrupting LAMININ‐mediated intercellular communication. SN‐011 may exert neuroprotective effects by targeting JUNB to upregulate ADGRE5 expression and restore the vEC‐centered cellular communication network, providing a theoretical basis for SN‐011 as a potential IS therapeutic.