title: Direct Intracranial Injection of AAVrh8 Encoding Monkey β-N-Acetylhexosaminidase Causes Neurotoxicity in the Primate Brain.
abstract: GM2 gangliosidoses, including Tay-Sachs disease and Sandhoff disease, are lysosomal storage disorders caused by deficiencies in β-N-acetylhexosaminidase (Hex). Patients are afflicted primarily with progressive central nervous system (CNS) dysfunction. Studies in mice, cats, and sheep have indicated safety and widespread distribution of Hex in the CNS after intracranial vector infusion of AAVrh8 vectors encoding species-specific Hex α- or β-subunits at a 1:1 ratio. Here, a safety study was conducted in cynomolgus macaques (cm), modeling previous animal studies, with bilateral infusion in the thalamus as well as in left lateral ventricle of AAVrh8 vectors encoding cm Hex α- and β-subunits. Three doses (3.2 × 1012vg [n = 3]; 3.2 × 1011vg [n = 2]; or 1.1 × 1011vg [n = 2]) were tested, with controls infused with vehicle (n = 1) or transgene empty AAVrh8 vector at the highest dose (n = 2). Most monkeys receiving AAVrh8-cmHexα/β developed dyskinesias, ataxia, and loss of dexterity, with higher dose animals eventually becoming apathetic. Time to onset of symptoms was dose dependent, with the highest-dose cohort producing symptoms within a month of infusion. One monkey in the lowest-dose cohort was behaviorally asymptomatic but had magnetic resonance imaging abnormalities in the thalami. Histopathology was similar in all monkeys injected with AAVrh8-cmHexα/β, showing severe white and gray matter necrosis along the injection track, reactive vasculature, and the presence of neurons with granular eosinophilic material. Lesions were minimal to absent in both control cohorts. Despite cellular loss, a dramatic increase in Hex activity was measured in the thalamus, and none of the animals presented with antibody titers against Hex. The high overexpression of Hex protein is likely to blame for this negative outcome, and this study demonstrates the variations in safety profiles of AAVrh8-Hexα/β intracranial injection among different species, despite encoding for self-proteins.
authors.affiliation: 1 Department of Neurology, University of Massachusetts Medical School , Worcester, Massachusetts.|2 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Diane
authors.initials: D
authors.lastname: Golebiowski
authors.affiliation: 3 Department of Radiology, University of Massachusetts Medical School , Worcester, Massachusetts.|4 New England Center for Stroke Research, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Imramsjah M J
authors.initials: IMJ
authors.lastname: van der Bom
authors.affiliation: 5 Department of Neurosurgery, Massachusetts General Hospital , Boston, Massachusetts.
authors.forename: Churl-Su
authors.initials: CS
authors.lastname: Kwon
authors.affiliation: 6 New England Primate Research Center, Harvard Medical School , Southborough, Massachusetts.
authors.forename: Andrew D
authors.initials: AD
authors.lastname: Miller
authors.affiliation: 6 New England Primate Research Center, Harvard Medical School , Southborough, Massachusetts.
authors.forename: Keiko
authors.initials: K
authors.lastname: Petrosky
authors.affiliation: 7 Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University , Alabama.|8 Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University , Alabama.
authors.forename: Allison M
authors.initials: AM
authors.lastname: Bradbury
authors.affiliation: 1 Department of Neurology, University of Massachusetts Medical School , Worcester, Massachusetts.|2 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Stacy
authors.initials: S
authors.lastname: Maitland
authors.affiliation: 3 Department of Radiology, University of Massachusetts Medical School , Worcester, Massachusetts.|4 New England Center for Stroke Research, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Anna Luisa
authors.initials: AL
authors.lastname: Kühn
authors.affiliation: 9 Department of Animal Medicine, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Nina
authors.initials: N
authors.lastname: Bishop
authors.affiliation: 6 New England Primate Research Center, Harvard Medical School , Southborough, Massachusetts.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Curran
authors.affiliation: 6 New England Primate Research Center, Harvard Medical School , Southborough, Massachusetts.
authors.forename: Nilsa
authors.initials: N
authors.lastname: Silva
authors.affiliation: 1 Department of Neurology, University of Massachusetts Medical School , Worcester, Massachusetts.|2 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Dwijit
authors.initials: D
authors.lastname: GuhaSarkar
authors.affiliation: 6 New England Primate Research Center, Harvard Medical School , Southborough, Massachusetts.
authors.forename: Susan V
authors.initials: SV
authors.lastname: Westmoreland
authors.affiliation: 7 Scott-Ritchey Research Center, College of Veterinary Medicine, Auburn University , Alabama.|8 Department of Anatomy, Physiology, and Pharmacology, College of Veterinary Medicine, Auburn University , Alabama.
authors.forename: Douglas R
authors.initials: DR
authors.lastname: Martin
authors.affiliation: 3 Department of Radiology, University of Massachusetts Medical School , Worcester, Massachusetts.|4 New England Center for Stroke Research, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Gounis
authors.affiliation: 10 Department of Neurosurgery, Alpert Medical School, Brown University , Providence, Rhode Island.|11 Brown Institute for Brain Science, Brown University , Providence, Rhode Island.|12 Rhode Island Hospital , Providence, Rhode Island.
authors.forename: Wael F
authors.initials: WF
authors.lastname: Asaad
authors.affiliation: 1 Department of Neurology, University of Massachusetts Medical School , Worcester, Massachusetts.|2 Horae Gene Therapy Center, University of Massachusetts Medical School , Worcester, Massachusetts.
authors.forename: Miguel
authors.initials: M
authors.lastname: Sena-Esteves
mesh_terms: D000818:Animals; D057565:Apathy; D000229:Dependovirus / Q000235:genetics* / Q000378:metabolism; D004195:Disease Models, Animal; D020820:Dyskinesias / Q000209:etiology* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D005260:Female; D020143:Gangliosidoses, GM2 / Q000235:genetics / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000009:adverse effects* / Q000737:chemistry / Q000378:metabolism; D066128:Gray Matter / Q000378:metabolism / Q000473:pathology; D007276:Injections, Intraventricular; D008252:Macaca fascicularis; D008297:Male; D009336:Necrosis / Q000209:etiology* / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D021122:Protein Subunits / Q000009:adverse effects / Q000235:genetics / Q000378:metabolism; D013788:Thalamus / Q000378:metabolism / Q000473:pathology; D019076:Transgenes; D066127:White Matter / Q000378:metabolism / Q000473:pathology; D001619:beta-N-Acetylhexosaminidases / Q000009:adverse effects / Q000235:genetics* / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D001619:beta-N-Acetylhexosaminidases
keywords: AAV; Tay-Sachs disease; adeno-associated virus; gene therapy; hexosaminidase; intracranial delivery