Abstract
Purpose :
Transcriptionally heterogeneous choroidal fibroblast subpopulations and their relationship to local microenvironmental signals are poorly understood. We aimed to investigate whether distinct fibroblast-like subpopulations exist within choroidal stromal cells by employing single-cell RNA sequencing on samples obtained from three unique human donors.
Methods :
Whole choroidal stromal cell (Ch-SC) cultures were established by collagenase digestion from three unique adult (aged 49-60 years) male human donors and analysed p5 using the 10X Genomics Chromium 3′ Gene Expression assay. The resulting data was processed using Galaxy Australia software, involving various quality control steps and analysis methods, including clustering based on the first 20 principal components from each cluster.
Results :
Analysis of 20,429 choroidal stromal cells revealed distinct clusters representing fibroblast subpopulations. Notably, clusters exhibited differential expression of genes associated with fibrosis, wound healing, and proliferation. For instance, cluster 0 showed expression of NEAT1 and NEAT2 promoting fibrosis, while cluster 1 exhibited significant expression of genes like S100A6 and TXN associated with fibroblast proliferation, morphology, and differentiation. Cluster 2 contained HLA-B and RBP1, indicating mesenchymal stromal cell characteristics and inhibition of fibroblast proliferation, respectively. Clusters 3, 4, and 5 were enriched with housekeeping genes. Significant inter-donor variation was also evident.
Conclusions :
Single-cell RNA sequencing identified distinct fibroblast subpopulations within a novel heterogenous choroidal stromal cell population. The transcriptome of Ch-SCs were enriched for genes associated with stromal cell proliferation and wound healing. These markers play a pivotal role in understanding the function of choroidal stromal cells.
This abstract was presented at the 2024 ARVO Annual Meeting, held in Seattle, WA, May 5-9, 2024.