Abstract
Mutations in the photoreceptor transcription factor gene cone-rod homeobox (CRX) lead to distinct retinopathy phenotypes, including early-onset vision impairment in dominant Leber congenital amaurosis (LCA). Using induced pluripotent stem cells (iPSCs) from a patient with CRX-I138fs48 mutation, we established an in vitro model of CRX-LCA in retinal organoids that showed defective photoreceptor maturation by histology and gene profiling, with diminished expression of visual opsins. Adeno-associated virus (AAV)-mediated CRX gene augmentation therapy partially restored photoreceptor phenotype and expression of phototransduction-related genes as determined by single-cell RNA-sequencing. Retinal organoids derived from iPSCs of a second dominant CRX-LCA patient carrying K88N mutation revealed the loss of opsin expression as a common phenotype, which was alleviated by AAV-mediated augmentation of CRX. Our studies provide a proof-of-concept for developing gene therapy of dominant CRX-LCA and other CRX retinopathies.
Original language | English |
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Pages (from-to) | 252-263 |
Number of pages | 12 |
Journal | Stem Cell Reports |
Volume | 16 |
Issue number | 2 |
Early online date | 28 Jan 2021 |
DOIs | |
Publication status | Published - 9 Feb 2021 |
Bibliographical note
Funding Information:We are grateful to the patients and their families for tissue donations, which enabled this study. We thank Jeanette Beers at the NIH/National Heart, Lung, and Blood Institute iPS and Genome Engineering Core Facility for generating iPSC lines, Sandra Burkett at the Molecular Cytogenetics Core Facility, National Cancer Institute Center for Cancer Research for karyotyping, and Milton A. English and Charles Drinnan for fibroblast cell line generation and stem cell maintenance. This research was supported by the Intramural Research Programs of the National Eye Institute ( ZIAEY000450 and ZIAEY000546 to A. Swaroop) and, in part, by the National Institute of Allergy and Infectious Diseases , National Institutes of Health. We are grateful to Opsomai Foundation (Italy) for a generous donation, which partly funded this work. This work used the high-performance computational capabilities of the NIH Biowulf Linux cluster ( http://hpc.nih.gov ).
Funding Information:
We are grateful to the patients and their families for tissue donations, which enabled this study. We thank Jeanette Beers at the NIH/National Heart, Lung, and Blood Institute iPS and Genome Engineering Core Facility for generating iPSC lines, Sandra Burkett at the Molecular Cytogenetics Core Facility, National Cancer Institute Center for Cancer Research for karyotyping, and Milton A. English and Charles Drinnan for fibroblast cell line generation and stem cell maintenance. This research was supported by the Intramural Research Programs of the National Eye Institute (ZIAEY000450 and ZIAEY000546 to A. Swaroop) and, in part, by the National Institute of Allergy and Infectious Diseases, National Institutes of Health. We are grateful to Opsomai Foundation (Italy) for a generous donation, which partly funded this work. This work used the high-performance computational capabilities of the NIH Biowulf Linux cluster (http://hpc.nih.gov).
Publisher Copyright:
© 2020 The Authors
Keywords
- pluripotent stem cells
- iPSC
- 3-D organoids
- retinal degeneration
- AAV
- therapy
- disease modeling
- transcription factor
- transcriptome
- scRNA-seq