Tie2‐Dependent Mechanisms Influence Leptomeningeal Collateral Dynamics and Reperfusion Following Stroke

Abstract Leptomeningeal collateral vessels help redistribute cerebral blood flow following arterial obstruction, reducing tissue damage. This study investigates the Tie2 receptor peptide agonist Vasculotide in a permanent middle cerebral artery occlusion (pMCAO) model. Vasculotide enhanced early diameter enlargement of pre‐existing pial collaterals, which may be mediated by structural remodeling, as evidenced by endothelial proliferation. These changes correlated with reduced infarct volume, blood‐brain barrier disruption, enhanced blood flow, and functional recovery at 3–28 days post‐pMCAO. Conditional endothelial cell (EC)‐specific EphA4 knockout (KO) mice exhibited increased Tie2 and Ang‐1 expression, mimicking the effects of Vasculotide on collateral size. Simultaneous genetic loss of EC‐specific EphA4 and Tie2 attenuated these outcomes. Nitric oxide inhibition partially blocked collateral enlargement in EC‐KO mice, suggesting the presence of additional contributors. Bulk RNAseq of meningeal tissue revealed upregulation of Krt5, Krt14, and Col17a1 in the ipsilateral meninges of Vasculotide‐treated and EC‐specific EphA4 KO mice. Notably, the number of Krt5‐expressing cells is increased on the leptomeningeal arterial vasculature of KO mice, suggesting a novel contribution to collateral enlargement. The opposing roles of EphA4 and Tie2 in collateral dynamics are demonstrated, and a novel molecular program is identified that can be targeted to enhance their diameter enlargement in ischemic stroke.