title: Metabolic correction in oligodendrocytes derived from metachromatic leukodystrophy mouse model by using encapsulated recombinant myoblasts.
abstract: In an effort to develop an encapsulated cell-based system to deliver arylsulfatase A (ARSA) to the central nervous system of metachromatic leukodystrophy (MLD) patients, we engineered C2C12 mouse myoblasts with a retroviral vector containing a full-length human ARSA cDNA and evaluated the efficacy of the recombinant secreted enzyme to revert the MLD phenotype in oligodendrocytes (OL) of the As2-/- mouse model. After transduction, C2C12 cells showed a fifteen-fold increase in intracellular ARSA activity and five-fold increase in ARSA secretion. The secreted hARSA collected from transduced cells encapsulated in polyether-sulfone polymer, was taken up by enzyme-deficient OL derived from MLD mice and normally sorted to the lysosomal compartment, where transferred enzyme reached 80% of physiological levels, restoring the metabolism of sulfatide. To evaluate whether secreted enzyme could restore metabolic function in the brain, encapsulated cells and secreted ARSA were shown to be stable in CSF in vitro. Further, to test cell viability and enzyme release in vivo, encapsulated cells were implanted subcutaneously on the dorsal flank of DBA/2J mice. One month later, all retrieved implants released hARSA at rates similar to unencapsulated cells and contained well preserved myoblasts, demonstrating that encapsulation maintains differentiation of C2C12 cells, stable transgene expression and long-term cell viability in vivo. Thus, these results show the promising potential of developing an ARSA delivery system to the CNS based on the use of a polymer-encapsulated transduced xenogenic cell line for gene therapy of MLD.
authors.affiliation: Telethon Institute for Gene Therapy (TIGET) and DIBIT, San Raffaele Scientific, Institute, Milan, Italy. aconsiglio@cmrb.eu
authors.forename: Antonella
authors.initials: A
authors.lastname: Consiglio
authors.forename: Sabata
authors.initials: S
authors.lastname: Martino
authors.forename: Diego
authors.initials: D
authors.lastname: Dolcetta
authors.forename: Gabriella
authors.initials: G
authors.lastname: Cusella
authors.forename: Massimo
authors.initials: M
authors.lastname: Conese
authors.forename: Sergio
authors.initials: S
authors.lastname: Marchesini
authors.forename: Giuliana
authors.initials: G
authors.lastname: Benaglia
authors.forename: Lawrence
authors.initials: L
authors.lastname: Wrabetz
authors.forename: Aldo
authors.initials: A
authors.lastname: Orlacchio
authors.forename: Nicole
authors.initials: N
authors.lastname: Déglon
authors.forename: Patrick
authors.initials: P
authors.lastname: Aebischer
authors.forename: Giovanni M
authors.initials: GM
authors.lastname: Severini
authors.forename: Claudio
authors.initials: C
authors.lastname: Bordignon
mesh_terms: D000818:Animals; D001192:Arylsulfatases / Q000235:genetics / Q000378:metabolism; D002214:Capsules / Q000627:therapeutic use; D002460:Cell Line; D002470:Cell Survival / Q000502:physiology; D004195:Disease Models, Animal; D005822:Genetic Vectors / Q000235:genetics*; D006085:Graft Survival / Q000502:physiology; D006801:Humans; D007966:Leukodystrophy, Metachromatic / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D051379:Mice; D018345:Mice, Knockout; D032446:Myoblasts / Q000201:enzymology / Q000637:transplantation*; D009416:Nerve Regeneration / Q000235:genetics; D009836:Oligodendroglia / Q000201:enzymology*; D011108:Polymers / Q000627:therapeutic use; D013433:Sulfoglycosphingolipids / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D019076:Transgenes / Q000235:genetics; D014183:Transplantation, Heterologous / Q000379:methods; D016896:Treatment Outcome; D015854:Up-Regulation / Q000235:genetics
chemical_list: D002214:Capsules; D011108:Polymers; D013433:Sulfoglycosphingolipids; D001192:Arylsulfatases