Krabbe disease is a rare neurodegenerative fatal disease. It is caused by deficiency of the lysosomal enzyme galactocerebrosidase (GALC), which results in progressive accumulation of galactolipid substrates in myelin-forming cells. However, there is still a lack of appropriate neural models and effective approaches for Krabbe disease. We generated induced pluripotent stem cells (iPSCs) from a Krabbe patient previously. Here, Krabbe patient-derived neural stem cells (K-NSCs) were induced from these iPSCs. By using nine kinds of recombinant adeno-associated virus (rAAV) vectors to infect K-NSCs, we found that the rAAV2 vector has high transduction efficiency for K-NSCs. Most importantly, rAAV2-GALC rescued GALC enzymatic activity in K-NSCs. Our findings not only establish a novel patient NSC model for Krabbe disease, but also firstly indicate the potential of rAAV2-mediated gene therapy for this devastating disease.
Krabbe disease is caused by the mutation or deficiency of galactocerebrosidase (GALC) enzyme, which is located in the lysosome and hydrolyzes the galactolipid substrates like psychosine. Psychosine would accumulate abnormally in the myelin forming cells and result in demyelination in the nervous systems with the clinical symptoms of spastic paraparesis and seizures. Adeno-associated virus (AAV) is a well-established and safe viral vector for gene delivery. However, effective AAV serotype for the transduction of the human neural stem cells (NSCs) has not been identified. Here, we screened a variety of AAV serotypes to transduce NSCs-related disease model induced by Krabbe patient induced pluripotent stem cells (iPSC) differentiation. It has been found that AAV2 has a higher transduction effect for NSCs, and AAV2 carrying GALC gene rescued the GALC enzymatic activity of Krabbe NSCs. Our findings established Krabbe patient iPSCs-derived NSCs as a new model for study the pathogenesis of Krabbe disease, and also demonstrated the potential of using AAV2 as a vector in gene therapy for Krabbe disease, which also proved the potential of AAV in devising gene therapy strategies for the treatment of genetic neurodegenerative diseases.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.