title: Pervasive supply of therapeutic lysosomal enzymes in the CNS of normal and Krabbe-affected non-human primates by intracerebral lentiviral gene therapy.
abstract: Metachromatic leukodystrophy (MLD) and globoid cell leukodystrophy (GLD or Krabbe disease) are severe neurodegenerative lysosomal storage diseases (LSD) caused by arylsulfatase A (ARSA) and galactosylceramidase (GALC) deficiency, respectively. Our previous studies established lentiviral gene therapy (GT) as a rapid and effective intervention to provide pervasive supply of therapeutic lysosomal enzymes in CNS tissues of MLD and GLD mice. Here, we investigated whether this strategy is similarly effective in juvenile non-human primates (NHP). To provide proof of principle for tolerability and biological efficacy of the strategy, we established a comprehensive study in normal NHP delivering a clinically relevant lentiviral vector encoding for the human ARSA transgene. Then, we injected a lentiviral vector coding for the human GALC transgene in Krabbe-affected rhesus macaques, evaluating for the first time the therapeutic potential of lentiviral GT in this unique LSD model. We showed favorable safety profile and consistent pattern of LV transduction and enzyme biodistribution in the two models, supporting the robustness of the proposed GT platform. We documented moderate inflammation at the injection sites, mild immune response to vector particles in few treated animals, no indication of immune response against transgenic products, and no molecular evidence of insertional genotoxicity. Efficient gene transfer in neurons, astrocytes, and oligodendrocytes close to the injection sites resulted in robust production and extensive spreading of transgenic enzymes in the whole CNS and in CSF, leading to supraphysiological ARSA activity in normal NHP and close to physiological GALC activity in the Krabbe NHP, in which biological efficacy was associated with preliminary indication of therapeutic benefit. These results support the rationale for the clinical translation of intracerebral lentiviral GT to address CNS pathology in MLD, GLD, and other neurodegenerative LSD.
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Vasco
authors.initials: V
authors.lastname: Meneghini
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Lattanzi
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Tiradani
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Bravo
authors.affiliation: Department of Chemistry, Biology and Biotechnologies, Biochemistry and Molecular Biology Unit, University of Perugia, Perugia, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Morena
authors.affiliation: Anatomy and Histopathology Department, San Raffaele Scientific Institute, Milano, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Sanvito
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Calabria
authors.affiliation: University of California San Francisco (UCSF), San Francisco, CA, USA.
authors.forename: John
authors.initials: J
authors.lastname: Bringas
authors.affiliation: Division of Regenerative Medicine, Tulane National Primate Research Center, Covington, LA, USA.
authors.forename: Jeanne M
authors.initials: JM
authors.lastname: Fisher-Perkins
authors.affiliation: Division of Regenerative Medicine, Tulane National Primate Research Center, Covington, LA, USA.
authors.forename: Jason P
authors.initials: JP
authors.lastname: Dufour
authors.affiliation: Division of Regenerative Medicine, Tulane National Primate Research Center, Covington, LA, USA.
authors.forename: Kate C
authors.initials: KC
authors.lastname: Baker
authors.affiliation: Anatomy and Histopathology Department, San Raffaele Scientific Institute, Milano, Italy Vita-Salute San Raffaele University, Milan, Italy.
authors.forename: Claudio
authors.initials: C
authors.lastname: Doglioni
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy.
authors.forename: Eugenio
authors.initials: E
authors.lastname: Montini
authors.affiliation: Division of Regenerative Medicine, Tulane National Primate Research Center, Covington, LA, USA.
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Bunnell
authors.affiliation: University of California San Francisco (UCSF), San Francisco, CA, USA.
authors.forename: Krystof
authors.initials: K
authors.lastname: Bankiewicz
authors.affiliation: Department of Chemistry, Biology and Biotechnologies, Biochemistry and Molecular Biology Unit, University of Perugia, Perugia, Italy.
authors.forename: Sabata
authors.initials: S
authors.lastname: Martino
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy Vita-Salute San Raffaele University, Milan, Italy.
authors.forename: Luigi
authors.initials: L
authors.lastname: Naldini
authors.affiliation: San Raffaele Telethon Institute for Gene Therapy (SR-TIGET), Division of Regenerative Medicine, Stem Cells and Gene Therapy, IRCCS San Raffaele Scientific Institute, Milan, Italy gritti.angela@hsr.it.
authors.forename: Angela
authors.initials: A
authors.lastname: Gritti
mesh_terms: D000818:Animals; D002553:Cerebroside-Sulfatase / Q000235:genetics* / Q000378:metabolism; D004195:Disease Models, Animal; D005698:Galactosylceramidase / Q000235:genetics* / Q000378:metabolism; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D007965:Leukodystrophy, Globoid Cell / Q000628:therapy*; D007966:Leukodystrophy, Metachromatic / Q000628:therapy*; D008253:Macaca mulatta; D051379:Mice; D014161:Transduction, Genetic; D016896:Treatment Outcome
chemical_list: D002553:Cerebroside-Sulfatase; D005698:Galactosylceramidase
keywords: brain; gene therapy; lentiviral vectors; leukodystrophy; non‐human primates