Iron oxide-coupled CRISPR-nCas9-based genome editing assessment in mucopolysaccharidosis IVA mice

Andrés Felipe Leal; Betul Celik; Nidhi Fnu; Shaukat Khan; Shunji Tomatsu; Carlos Javier Alméciga-Díaz

doi:10.1016/j.omtm.2023.101153

Iron oxide-coupled CRISPR-nCas9-based genome editing assessment in mucopolysaccharidosis IVA mice

Mol Ther Methods Clin Dev. 2023 Nov 7:31:101153. doi: 10.1016/j.omtm.2023.101153. eCollection 2023 Dec 14.

Authors

Andrés Felipe Leal^{1

2}, Betul Celik^{2

3}, Nidhi Fnu^{2

3}, Shaukat Khan², Shunji Tomatsu^{2

3

4

5}, Carlos Javier Alméciga-Díaz¹

Affiliations

¹ Institute for the Study of Inborn Errors of Metabolism, Faculty of Science, Pontificia Universidad Javeriana, Bogotá DC 110231, Colombia.
² Nemours Children's Health, Wilmington, DE 19803, USA.
³ Faculty of Arts and Sciences, University of Delaware, Newark, DE 19716, USA.
⁴ Department of Pediatrics, Graduate School of Medicine, Gifu University, Gifu 501-1193, Japan.
⁵ Department of Pediatrics, Thomas Jefferson University, Philadelphia, PA 19144, USA.

Abstract

Mucopolysaccharidosis (MPS) IVA is a lysosomal storage disorder caused by mutations in the GALNS gene that leads to the lysosomal accumulation of keratan sulfate (KS) and chondroitin 6-sulfate, causing skeletal dysplasia and cardiopulmonary complications. Current enzyme replacement therapy does not impact the bone manifestation of the disease, supporting that new therapeutic alternatives are required. We previously demonstrated the suitability of the CRISPR-nCas9 system to rescue the phenotype of human MPS IVA fibroblasts using iron oxide nanoparticles (IONPs) as non-viral vectors. Here, we have extended this strategy to an MPS IVA mouse model by inserting the human GALNS cDNA into the ROSA26 locus. The results showed increased GALNS activity, mono-KS reduction, partial recovery of the bone pathology, and non-IONPs-related toxicity or antibody-mediated immune response activation. This study provides, for the first time, in vivo evidence of the potential of a CRISPR-nCas9-based gene therapy strategy for treating MPS IVA using non-viral vectors as carriers.

Keywords: CRISPR-nCas9; MPS IVA; genome editing; non-viral vectors.