Language
English
Publication Date
3-14-2024
Journal
Molecular Therapy Methods & Clinical Development
DOI
10.1016/j.omtm.2024.101208
PMID
38414825
PMCID
PMC10897892
PubMedCentral® Posted Date
2-6-2024
PubMedCentral® Full Text Version
Post-print
Abstract
Wiskott-Aldrich syndrome (WAS) is a severe X-linked primary immunodeficiency resulting from a diversity of mutations distributed across all 12 exons of the WAS gene. WAS encodes a hematopoietic-specific and developmentally regulated cytoplasmic protein (WASp). The objective of this study was to develop a gene correction strategy potentially applicable to most WAS patients by employing nuclease-mediated, site-specific integration of a corrective WAS gene sequence into the endogenous WAS chromosomal locus. In this study, we demonstrate the ability to target the integration of WAS2-12-containing constructs into intron 1 of the endogenous WAS gene of primary CD34+ hematopoietic stem and progenitor cells (HSPCs), as well as WASp-deficient B cell lines and WASp-deficient primary T cells. This intron 1 targeted integration (TI) approach proved to be quite efficient and restored WASp expression in treated cells. Furthermore, TI restored WASp-dependent function to WAS patient T cells. Edited CD34+ HSPCs exhibited the capacity for multipotent differentiation to various hematopoietic lineages in vitro and in transplanted immunodeficient mice. This methodology offers a potential editing approach for treatment of WAS using patient’s CD34+ cells.
Keywords
gene editing, gene therapy, primary immune disease, Wiskott-Aldrich syndrome, immunology
Published Open-Access
yes
Recommended Citation
Pille, Melissa; Avila, John M; Park, So Hyun; et al., "Gene Editing-Based Targeted Integration for Correction of Wiskott-Aldrich Syndrome" (2024). The Brown Foundation: Institute of Molecular Medicine. 68.
https://digitalcommons.library.tmc.edu/molecular_med/68
Graphical Abstract