CXCR4⁺ monocytes promote neovascularization in wet age-related macular degeneration

Wet age-related macular degeneration (wAMD) pathogenesis is primarily driven by choroidal neovascularization (CNV) mediated through intricate angiogenesis-inflammation crosstalk. While chemokine signaling has emerged as a promising therapeutic target, the specific cellular mediators orchestrating this process remain poorly defined. In this study, we employed an integrative approach combining bulk RNA sequencing, single-cell transcriptomics, multiplex immunohistochemistry (mIHC), and in vivo models to systematically elucidate the pivotal role of chemokine signaling in wAMD-associated angiogenesis and identify critical monocyte subsets involved in neovascularization. Our comprehensive analysis revealed MMP2+ endothelial cells (ECs) as a distinct cellular subset demonstrating dual angiogenic and stem-like properties, originating from a dysregulated endothelial differentiation pathway. Furthermore, we identified CXCR4+ monocytes as crucial mediators of pathological angiogenesis, with their activity likely modulated through CXCL12-CXCR4 chemokine axis interactions with MMP2+ ECs. Through transcription factor network analysis, we established RUNX3 as a master regulatory element governing pro-angiogenic monocyte differentiation, providing novel mechanistic insights into monocyte-driven angiogenesis in wAMD pathogenesis. These findings collectively establish MMP2+ ECs and CXCR4+ monocyte signaling as central pathogenic drivers in wAMD. Our results propose that therapeutic targeting of RUNX3-mediated chemokine signaling could synergize effectively with current anti-VEGF treatment paradigms.