Reimagining VEGF Therapy: The Role of PLCγ2 in Pan-Organ Endothelial Networks

Abstract

Objective: Ischemic tissue perfusion remains a major clinical challenge despite VEGF’s central role in angiogenesis. Limited success of VEGF-based therapies underscores the need to understand endothelial heterogeneity, as distinct subsets exhibit differential responsiveness. This study explores conserved endothelial states that govern VEGF-driven angiogenesis across human organs and evaluate their functional significance. Approach: We performed single-cell and single-nucleus RNA sequencing (scRNA-seq/snRNA-seq) analysis of endothelial cells from eight human tissues using GTEx data, validated findings in independent skin and heart datasets, and conducted CRISPR-mediated PLCγ2 knockout in cultured endothelial cells followed by scRNA-seq. Results: Analysis of 209,126 nuclei revealed a universal arterial-like endothelial subset (Cluster 2) enriched across all organs, characterized by high expression of PLCγ2, a critical VEGF signaling effector. PLCγ2hi cells exhibited robust angiogenic and lymphangiogenic programs, including upregulation of VEGFC, NOTCH4, and JAG1, and enrichment of pathways for vasculature development and endothelial differentiation. Cell–cell communication analysis demonstrated exclusive BMP6, IGF2, and CXCL12 signaling from PLCγ2hi cells to fibroblasts, pericytes, and immune cells, positioning this subset as a central angiogenic hub. Functional validation showed that PLCγ2 knockout depleted PLCγ2hi clusters, suppressed VEGF signaling, and diminished BMP6 ligand secretion, confirming its role in maintaining pro-angiogenic endothelial states. Innovation: Ineffective targeting of the appropriate endothelial cell state may limit current therapeutic VEGF strategies, positioning PLCγ2hi endothelial cells as a clinically relevant target. Conclusion: We identify PLCγ2hi arterial endothelial cells as a conserved pan-organ population critical for VEGF responsiveness and vascular regeneration. These findings reimagine VEGF therapy by highlighting PLCγ2 as a critical regulator of a pro-angiogenic arterial-like endothelial state and a promising combinatorial target to overcome limitations of current angiogenic strategies.