Abstract
The management of preretinal fibrovascular membranes, a devastating complication of advanced diabetic retinopathy (DR), remains challenging. We characterized the molecular profile of cell populations in these fibrovascular membranes to identify potentially new therapeutic targets. Preretinal fibrovascular membranes were surgically removed from patients and submitted for single-cell RNA-Seq (scRNA-Seq). Differential gene expression was implemented to define the transcriptomics profile of these cells and revealed the presence of endothelial, inflammatory, and stromal cells. Endothelial cell reclustering identified subclusters characterized by noncanonical transcriptomics profile and active angiogenesis. Deeper investigation of the inflammatory cells showed a subcluster of macrophages expressing proangiogenic cytokines, presumably contributing to angiogenesis. The stromal cell cluster included a pericyte-myofibroblast transdifferentiating subcluster, indicating the involvement of pericytes in fibrogenesis. Differentially expressed gene analysis showed that Adipocyte Enhancer-binding Protein 1, AEBP1, was significantly upregulated in myofibroblast clusters, suggesting that this molecule may have a role in transformation. Cell culture experiments with human retinal pericytes (HRP) in high-glucose condition confirmed the molecular transformation of pericytes toward myofibroblastic lineage. AEBP1 siRNA transfection in HRP reduced the expression of profibrotic markers in high glucose. In conclusion, AEBP1 signaling modulates pericyte-myofibroblast transformation, suggesting that targeting AEBP1 could prevent scar tissue formation in advanced DR.
| Original language | English (US) |
|---|---|
| Article number | e172062 |
| Journal | JCI Insight |
| Volume | 8 |
| Issue number | 23 |
| DOIs | |
| State | Published - 2023 |
Funding
Imaging work was performed at the Northwestern University Center for Advanced Microscopy, generously supported by NCI CCSG P30 CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. We would like to thank Yi-Wen Hsieh for his help in imaging immunofluorescence experiments, Matthew Schipma for running our raw sequencing data through the 10x Genomics Cell Ranger 4.0.0 pipeline, and Luisa Iruela Arispe for helpful discussions about the manuscript. The present work is supported by the following grants: NIH-R01-EY30121-A1 and Juvenile Diabetes Research Foundation Grant INO-2022-1112-A-N (AAF). JAL was supported by NIH grant K08 EY030923, NIH grant R01 EY034486, and the Research to Prevent Blindness Sybil B. Harrington Career Development Award for Macular Degeneration. BRT was supported by NIH R01 EY032609 and Brightfocus Foundation new investigator grant in macular degeneration research M2021018N.
ASJC Scopus subject areas
- General Medicine
