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Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells

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Published on: May 22, 2017

Single-cell RNA sequencing reveals a unique pericyte type associated with capillary dysfunction.

Min Xia^1,2, Lyu Jiao³, Xiao-Han Wang³

¹The Fourth School of Clinical Medicine, Nanjing Medical University, Nanjing 210000, China.

|May 22, 2023

Summary

Collagen type 1 alpha 1 (Col1a1) is a key marker for vulnerable pericytes in retinal capillary dysfunction. Targeting Col1a1 shows promise for treating proliferative retinopathies.

Keywords:

Capillary dysfunction Pericyte Pericyte-myofibroblast transition Single-cell RNA sequencing

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Single-cell RNA Sequencing and Analysis of Human Pancreatic Islets

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Area of Science:

Vascular biology
Retinal cell biology
Molecular ophthalmology

Background:

Capillary dysfunction, marked by pericyte and endothelial cell (EC) degeneration, underlies severe vascular diseases.
The molecular mechanisms driving pericyte heterogeneity and their role in capillary dysfunction remain incompletely understood.

Purpose of the Study:

To elucidate the molecular profiles governing pericyte heterogeneity in the context of retinal capillary dysfunction.
To identify specific pericyte subpopulations and molecular targets for therapeutic intervention in conditions like oxygen-induced proliferative retinopathy (OIR).

Main Methods:

Single-cell RNA sequencing of mouse retinas to create a comprehensive cellular atlas.
Bioinformatics analysis, including sub-clustering, Gene Ontology (GO), and KEGG pathway analysis, to identify vulnerable pericyte populations.
Quantitative reverse transcription PCR (qRT-PCR), Western blotting, and in vitro assays (Matrigel co-culture, PI, JC-1 staining) to validate the role of Collagen type 1 alpha 1 (Col1a1).

Main Results:

A single-cell atlas revealed 10 distinct retinal cell types, with pericytes sub-clustered into three subpopulations.
Pericyte sub-population 2 was identified as vulnerable to capillary dysfunction, with Col1a1 identified as its marker gene.
Col1a1 expression was upregulated in OIR retinas and in patients with proliferative diabetic retinopathy (PDR) and retinopathy of prematurity (ROP); Col1a1 silencing ameliorated OIR pathology in vitro and in vivo.

Conclusions:

Col1a1 is a critical marker for a vulnerable pericyte subpopulation and a potential therapeutic target for retinal capillary dysfunction.
Targeting Col1a1 may offer a novel strategy for treating sight-threatening proliferative retinopathies by mitigating pericyte loss and neovascularization.