Gene Therapy of Dominant CRX-Leber Congenital Amaurosis using Patient Stem Cell-Derived Retinal Organoids.
Adult
Cell Differentiation
Child
Child, Preschool
Dependovirus
Female
Genetic Therapy
/ methods
Homeodomain Proteins
/ genetics
Humans
Induced Pluripotent Stem Cells
/ cytology
Leber Congenital Amaurosis
/ genetics
Models, Biological
Mutation
Opsins
/ metabolism
Organoids
/ cytology
Phenotype
Photoreceptor Cells
/ metabolism
Retina
/ cytology
Sequence Analysis, RNA
Single-Cell Analysis
Trans-Activators
/ genetics
Transcriptome
3-D organoids
AAV
disease modeling
iPSC
pluripotent stem cells
retinal degeneration
scRNA-seq
therapy
transcription factor
transcriptome
Journal
Stem cell reports
ISSN: 2213-6711
Titre abrégé: Stem Cell Reports
Pays: United States
ID NLM: 101611300
Informations de publication
Date de publication:
09 02 2021
09 02 2021
Historique:
received:
13
08
2020
revised:
27
12
2020
accepted:
28
12
2020
pubmed:
30
1
2021
medline:
30
11
2021
entrez:
29
1
2021
Statut:
ppublish
Résumé
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.
Identifiants
pubmed: 33513359
pii: S2213-6711(20)30513-0
doi: 10.1016/j.stemcr.2020.12.018
pmc: PMC7878833
pii:
doi:
Substances chimiques
Homeodomain Proteins
0
Opsins
0
Trans-Activators
0
cone rod homeobox protein
0
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Langues
eng
Sous-ensembles de citation
IM
Pagination
252-263Subventions
Organisme : Intramural NIH HHS
ID : ZIA EY000450
Pays : United States
Organisme : Intramural NIH HHS
ID : ZIA EY000546
Pays : United States
Informations de copyright
Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.
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