Design of a regulated lentiviral vector for hematopoietic stem cell gene therapy of globoid cell leukodystrophy.
Globoid cell leukodystrophy (GLD) is a demyelinating lysosomal storage disease due to the deficiency of the galactocerebrosidase (GALC) enzyme. The favorable outcome of hematopoietic stem and progenitor cell (HSPC)-based approaches in GLD and other similar diseases suggests HSPC gene therapy as a promising therapeutic option for patients. The path to clinical development of this strategy was hampered by a selective toxicity of the overexpressed GALC in the HSPC compartment. Here, we presented the optimization of a lentiviral vector (LV) in which miR-126 regulation was coupled to codon optimization of the human GALC cDNA to obtain a selective and enhanced enzymatic activity only upon transduced HSPCs differentiation. The safety of human GALC overexpression driven by this LV was extensively demonstrated in vitro and in vivo on human HSPCs from healthy donors. No perturbation in the content of proapoptotic sphingolipids, gene expression profile, and capability of engraftment and mutlilineage differentiation in chimeric mice was observed. The therapeutic potential of this LV was then assessed in a severe GLD murine model that benefited from transplantation of corrected HSPCs with longer survival and ameliorated phenotype as compared to untreated siblings. This construct has thus been selected as a candidate for clinical translation.
San Raffaele Telethon Institute for Gene Therapy , San Raffaele Scientific Institute , Milan, Italy.
Silvia
S
Ungari
San Raffaele Telethon Institute for Gene Therapy , San Raffaele Scientific Institute , Milan, Italy.
Annita
A
Montepeloso
Department of Experimental Medicine and Biochemical Science, Perugia University , Italy.
Francesco
F
Morena
Center for Regenerative Medicine "Stefano Ferrari," Department of Life Sciences, University of Modena and Reggio Emilia , Modena, Italy.
Fabienne
F
Cocchiarella
Center for Regenerative Medicine "Stefano Ferrari," Department of Life Sciences, University of Modena and Reggio Emilia , Modena, Italy.
Alessandra
A
Recchia
Department of Experimental Medicine and Biochemical Science, Perugia University , Italy.
Sabata
S
Martino
San Raffaele Telethon Institute for Gene Therapy , San Raffaele Scientific Institute , Milan, Italy.
Bernhard
B
Gentner
San Raffaele Telethon Institute for Gene Therapy , San Raffaele Scientific Institute , Milan, Italy ; Vita-Salute San Raffaele University , Milan, Italy.
Luigi
L
Naldini
San Raffaele Telethon Institute for Gene Therapy , San Raffaele Scientific Institute , Milan, Italy ; Vita-Salute San Raffaele University , Milan, Italy.
Alessandra
A
Biffi