Selection of Adeno-Associated Virus Serotypes For Gene Therapy In Krabbe Patient IPSCs-Derived NSCs

Abstract

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.