Gene Therapy of Dominant CRX-Leber Congenital Amaurosis using Patient Stem Cell-Derived Retinal Organoids

Kamil Kruczek, Zepeng Qu, James Gentry, Benjamin R Fadl, Linn Gieser, Suja Hiriyanna, Zachary Batz, Mugdha Samant, Ananya Samanta, Colin J Chu, Laura Campello, Brian P Brooks, Zhijian Wu, Anand Swaroop

Research output: Contribution to journalArticle (Academic Journal)peer-review

10 Citations (Scopus)
66 Downloads (Pure)

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 languageEnglish
Pages (from-to)252-263
Number of pages12
JournalStem Cell Reports
Volume16
Issue number2
Early online date28 Jan 2021
DOIs
Publication statusPublished - 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

Fingerprint

Dive into the research topics of 'Gene Therapy of Dominant CRX-Leber Congenital Amaurosis using Patient Stem Cell-Derived Retinal Organoids'. Together they form a unique fingerprint.

Cite this