Abstract
Patients with neovascular AMD (nAMD) suffer vision loss from destructive angiogenesis, termed choroidal neovascularization (CNV). Macrophages are found in CNV lesions from patients with nAMD. Additionally, Ccr2-/- mice, which lack classical monocyte-derived macrophages, show reduced CNV size. However, macrophages are highly diverse cells that can perform multiple functions. We performed single-cell RNA-Seq on immune cells from WT and Ccr2-/- eyes to uncover macrophage heterogeneity during the laser-induced CNV mouse model of nAMD. We identified 12 macrophage clusters, including Spp1+ macrophages. Spp1+ macrophages were enriched from WT lasered eyes and expressed a proangiogenic transcriptome via multiple pathways, including vascular endothelial growth factor signaling, endothelial cell sprouting, cytokine signaling, and fibrosis. Additionally, Spp1+ macrophages expressed the marker CD11c, and CD11c+ macrophages were increased by laser and present in CNV lesions. Finally, CD11c+ macrophage depletion reduced CNV size by 40%. These findings broaden our understanding of ocular macrophage heterogeneity and implicate CD11c+ macrophages as potential therapeutic targets for treatment-resistant patients with nAMD.
| Original language | English (US) |
|---|---|
| Article number | e168142 |
| Journal | JCI Insight |
| Volume | 8 |
| Issue number | 7 |
| DOIs | |
| State | Published - 2023 |
Funding
We thank Matthew Schipma for running our raw sequencing data through the 10X Genomics Cell Ranger 3.1.0 pipeline. CMC was supported by a Lupus Research Alliance Novel Research Grant, a Rheumatology Research Foundation Innovative Research Award, and a Northwestern University Dixon Translational Research Grant Initiative Award. HP was supported by NIH grants AR064546, HL134375, AG049665, and UH2AR067687; the United States-Israel Binational Science Foundation (2013247); and the Rheumatology Research Foundation (Agmt 05/06/14). HP was also supported by the Mabel Greene Myers Professor of Medicine and by generous donations to the Rheumatology Precision Medicine Fund. JAL was supported by NIH grant K08 EY030923 and the Research to Prevent Blindness Sybil B. Harrington Career Development Award for Macular Degeneration. This study was supported by an Unrestricted Departmental Grant from Research to Prevent Blindness. Imaging work was performed at the Northwestern University Center for Advanced Microscopy, generously supported by CCSG P30 CA060553, awarded to the Robert H. Lurie Comprehensive Cancer Center. Flow cytometry was performed at the Northwestern University - Flow Cytometry Core Facility supported by Cancer Center Support Grant (NCI CA060553). scRNA-Seq was performed by the Northwestern University Metabolomics Core for “Integrative Genomics” and the NUSeq Core Facility. No funding body had any role in the design of the study, collection of data, data analysis, interpretation of data, or writing of the manuscript.
ASJC Scopus subject areas
- General Medicine