Single-cell transcriptomics of the human retinal pigment epithelium and choroid in health and macular degeneration

Andrew P Voigt; Kelly Mulfaul; Nathaniel K Mullin; Miles J Flamme-Wiese; Joseph C Giacalone; Edwin M Stone; Budd A Tucker; Todd E Scheetz; Robert F Mullins

doi:10.1073/pnas.1914143116

Back

Single-cell transcriptomics of the human retinal pigment epithelium and choroid in health and macular degeneration

Journal article

Open access

Peer reviewed

Single-cell transcriptomics of the human retinal pigment epithelium and choroid in health and macular degeneration

Andrew P Voigt, Kelly Mulfaul, Nathaniel K Mullin, Miles J Flamme-Wiese, Joseph C Giacalone, Edwin M Stone, Budd A Tucker, Todd E Scheetz and Robert F Mullins

Proceedings of the National Academy of Sciences - PNAS, Vol.116(48), pp.24100-24107

11/26/2019

DOI: 10.1073/pnas.1914143116

PMCID: PMC6883845

PMID: 31712411

Files and links (1)

url

https://doi.org/10.1073/pnas.1914143116View

Published (Version of record) Open Access

Abstract

The retinal pigment epithelium and the choroid are complex tissues whose dysfunction can lead to irreversible visual loss. In this study, single-cell RNA sequencing of both of these tissues was performed to characterize gene expression patterns specific to the retinal pigment epithelium and all major choroidal cell populations. Unique gene expression signatures of arterial, venous, and choriocapillaris vascular beds within the choroid were identified. RGCC , a gene that responds to complement and has been shown to induce endothelial apoptosis, was specifically expressed in choriocapillaris endothelial cells. This study provides potential insight into the molecular mechanisms of choroidal vascular disease and its contribution to age-related macular degeneration. The human retinal pigment epithelium (RPE) and choroid are complex tissues that provide crucial support to the retina. Disease affecting either of these supportive tissues can lead to irreversible blindness in the setting of age-related macular degeneration. In this study, single-cell RNA sequencing was performed on macular and peripheral regions of RPE-choroid from 7 human donor eyes in 2 independent experiments. In the first experiment, total RPE/choroid preparations were evaluated and expression profiles specific to RPE and major choroidal cell populations were identified. As choroidal endothelial cells represent a minority of the total RPE/choroidal cell population but are strongly implicated in age-related macular degeneration (AMD) pathogenesis, a second single-cell RNA-sequencing experiment was performed using endothelial cells enriched by magnetic separation. In this second study, we identified gene expression signatures along the choroidal vascular tree, classifying the transcriptome of human choriocapillaris, arterial, and venous endothelial cells. We found that the choriocapillaris highly and specifically expresses the regulator of cell cycle gene ( RGCC ), a gene that responds to complement activation and induces apoptosis in endothelial cells. In addition, RGCC was the most up-regulated choriocapillaris gene in a donor diagnosed with AMD. These results provide a characterization of the human RPE and choriocapillaris transcriptome, offering potential insight into the mechanisms of choriocapillaris response to complement injury and choroidal vascular disease in age-related macular degeneration.

choroid

Biological Sciences

single cell

age-related macular degeneration

choriocapillaris

Details

Title: Subtitle: Single-cell transcriptomics of the human retinal pigment epithelium and choroid in health and macular degeneration
Creators: Andrew P Voigt - Department of Ophthalmology and Visual Sciences
Kelly Mulfaul - Department of Ophthalmology and Visual Sciences
Nathaniel K Mullin - Department of Ophthalmology and Visual Sciences
Miles J Flamme-Wiese - Department of Ophthalmology and Visual Sciences
Joseph C Giacalone - Department of Ophthalmology and Visual Sciences
Edwin M Stone - Department of Ophthalmology and Visual Sciences
Budd A Tucker - Department of Ophthalmology and Visual Sciences
Todd E Scheetz - Department of Ophthalmology and Visual Sciences
Robert F Mullins - Department of Ophthalmology and Visual Sciences
Resource Type: Journal article
Publication Details: Proceedings of the National Academy of Sciences - PNAS, Vol.116(48), pp.24100-24107
DOI: 10.1073/pnas.1914143116
PMID: 31712411
PMCID: PMC6883845
NLM abbreviation: Proc Natl Acad Sci U S A
ISSN: 0027-8424
eISSN: 1091-6490
Publisher: National Academy of Sciences
Grant note: EY024605 / HHS | NIH | National Eye Institute (NEI) EY025580 / HHS | NIH | National Eye Institute (NEI) GM007337 / HHS | NIH | National Institute of General Medical Sciences (NIGMS) EY027038 / HHS | NIH | National Eye Institute (NEI)
Language: English
Date published: 11/26/2019
Academic Unit: Roy J. Carver Department of Biomedical Engineering; Electrical and Computer Engineering; The University of Iowa Institute for Vision Research; Iowa Neuroscience Institute; John and Marcia Carver Nonprofit Genetic Testing Laboratory; Ophthalmology and Visual Sciences
Record Identifier: 9984070287302771

Metrics

19 Record Views

277 Times Cited - Web of Science