title: A GLP-Compliant Toxicology and Biodistribution Study: Systemic Delivery of an rAAV9 Vector for the Treatment of Mucopolysaccharidosis IIIB.
abstract: No treatment is currently available for mucopolysaccharidosis (MPS) IIIB, a neuropathic lysosomal storage disease due to defect in α-N-acetylglucosaminidase (NAGLU). In preparation for a clinical trial, we performed an IND-enabling GLP-toxicology study to assess systemic rAAV9-CMV-hNAGLU gene delivery in WT C57BL/6 mice at 1 × 10(14) vg/kg and 2 × 10(14) vg/kg (n = 30/group, M:F = 1:1), and non-GLP testing in MPS IIIB mice at 2 × 10(14) vg/kg. Importantly, no adverse clinical signs or chronic toxicity were observed through the 6 month study duration. The rAAV9-mediated rNAGLU expression was rapid and persistent in virtually all tested CNS and somatic tissues. However, acute liver toxicity occurred in 33% (5/15) WT males in the 2 × 10(14) vg/kg cohort, which was dose-dependent, sex-associated, and genotype-specific, likely due to hepatic rNAGLU overexpression. Interestingly, a significant dose response was observed only in the brain and spinal cord, whereas in the liver at 24 weeks postinfection (pi), NAGLU activity was reduced to endogenous levels in the high dose cohort but remained at supranormal levels in the low dose group. The possibility of rAAV9 germline transmission appears to be minimal. The vector delivery resulted in transient T-cell responses and characteristic acute antibody responses to both AAV9 and rNAGLU in all rAAV9-treated animals, with no detectable impacts on tissue transgene expression. This study demonstrates a generally safe and effective profile, and may have identified the upper dosing limit of rAAV9-CMV-hNAGLU via systemic delivery for the treatment of MPS IIIB.
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Aaron S
authors.initials: AS
authors.lastname: Meadows
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: F Jason
authors.initials: FJ
authors.lastname: Duncan
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Marybeth
authors.initials: M
authors.lastname: Camboni
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Kathryn
authors.initials: K
authors.lastname: Waligura
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Chrystal
authors.initials: C
authors.lastname: Montgomery
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Kimberly
authors.initials: K
authors.lastname: Zaraspe
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Bartholomew J
authors.initials: BJ
authors.lastname: Naughton
authors.affiliation: 2 Center for Vaccines and Immunity, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: William G
authors.initials: WG
authors.lastname: Bremer
authors.affiliation: 3 Drug and Device Development Service, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Christopher
authors.initials: C
authors.lastname: Shilling
authors.affiliation: 2 Center for Vaccines and Immunity, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.|4 Department of Pediatrics, College of Medicine and Public Health, Columbus, Ohio.
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Walker
authors.affiliation: 5 Comparative Pathology and Mouse Phenotyping Shared Resource, Comprehensive Cancer Center, The Ohio State University, Columbus, Ohio.
authors.forename: Brad
authors.initials: B
authors.lastname: Bolon
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.|4 Department of Pediatrics, College of Medicine and Public Health, Columbus, Ohio.
authors.forename: Kevin M
authors.initials: KM
authors.lastname: Flanigan
authors.affiliation: 4 Department of Pediatrics, College of Medicine and Public Health, Columbus, Ohio.|6 Center for Cardiovascular and Pulmonary Research, Columbus, Ohio.
authors.forename: Kim L
authors.initials: KL
authors.lastname: McBride
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.|4 Department of Pediatrics, College of Medicine and Public Health, Columbus, Ohio.
authors.forename: Douglas M
authors.initials: DM
authors.lastname: McCarty
authors.affiliation: 1 Center for Gene Therapy, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.|3 Drug and Device Development Service, The Research Institute at Nationwide Children's Hospital, Columbus, Ohio.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Fu
mesh_terms: D000118:Acetylglucosaminidase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics / Q000378:metabolism; D015316:Genetic Therapy / Q000009:adverse effects* / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage / Q000009:adverse effects*; D008099:Liver / Q000378:metabolism*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009084:Mucopolysaccharidosis III / Q000628:therapy*; D009928:Organ Specificity; D017410:Practice Guidelines as Topic; D013116:Spinal Cord / Q000378:metabolism*
chemical_list: C033079:alpha-N-acetyl-D-glucosaminidase; D000118:Acetylglucosaminidase