Orai1 Facilitates Angiogenesis After Myocardial Infarction Through Notch1 Signaling Pathway

Post-ischemic neovascularization is crucial for cardiac repair after myocardial infarction (MI). While Orai1-dependent store-operated calcium entry is known to support angiogenesis, its specific role after MI remains unclear.

We stimulated human umbilical vein endothelial cells (ECs) with serum from patients with ST-segment-elevation MI to analyze proangiogenic mechanisms and to mimic the post-infarct systemic environment. We performed integrative analysis, including transcriptomics, proteomics, post-MI mouse heart single-cell RNA sequencing, and immunostaining.

ST-segment-elevation MI serum enhanced angiogenesis by upregulating VEGF (vascular endothelial growth factor), Notch, and Ca²⁺ signaling pathways in EC. Notably, it increased Orai1 expression and store-operated calcium entry activity, required for EC migration and proliferation. Consistently, Orai1 inhibition with CM4620 significantly impaired subintestinal venous plexus development in zebrafish embryos. Single-cell RNA sequencing confirmed Orai1 upregulation, particularly in tip cells and proliferating EC clusters, which was confirmed in peri-infarct regions of mouse hearts and in tip-like cells in a 3-dimensional culture model. Proteomics analysis revealed that Orai1 silencing dysregulated VEGF and Notch1-related proangiogenic proteins. Furthermore, IL (interleukin)-17A mimicked ST-segment-elevation MI serum, inducing Orai1-mediated store-operated calcium entry and EC migration.

Together, these findings reveal a novel role for the Orai1-dependent mechanism in post-MI angiogenesis, highlighting Orai1 as a potential therapeutic target for cardiac repair.