title: Adeno-associated virus-mediated aspartoacylase gene transfer to the brain of knockout mouse for canavan disease.
abstract: Canavan disease (CD) is an autosomal recessive leukodystrophy caused by deficiency of aspartoacylase (ASPA). Deficiency of ASPA leads to elevation of N-acetyl-L-aspartic acid (NAA) in the brain and urine. To explore the feasibility of gene transfer to replace ASPA in CD, we generated a knockout mouse and constructed an AAV vector that encodes human ASPA cDNA (hASPA) followed by green fluorescent protein (GFP) after an intraribosomal entry site. We injected CD mice with rAAV-hASPA-GFP in the striatum and thalamus or injected rAAV-GFP identically into control animals. Three to five months after the injection, we determined the presence of ASPA in the CD mouse brain by ASPA activity assay, GFP expression, and Western blot analysis. While rAAV-GFP-injected animals displayed undetectable levels of ASPA, all detection methods revealed significant ASPA levels in rAAV-hASPA-GFP-injected CD mice. We evaluated the functional effects of rAAV-hASPA-GFP-mediated ASPA expression by standard histological methods, magnetic resonance spectroscopy (MRS) for in vivo NAA levels, and magnetic resonance imaging of CD mice. rAAV-hASPA-injected animals displayed a remarkable lack of spongiform degeneration in the thalamus. However, pathology in sites unrelated to the injected areas showed no improvement in histopathology. The improvement in thalamic neuropathology was also detectable via in vivo MRI. MRS revealed that in vivo NAA levels were also reduced. These data indicate that rAAV-mediated ASPA delivery may be an interesting avenue for the treatment of CD.
authors.affiliation: Department of Pediatrics, Galveston, Texas 77555-0359, USA. rmatalon@utmb.edu
authors.forename: Reuben
authors.initials: R
authors.lastname: Matalon
authors.forename: Sankar
authors.initials: S
authors.lastname: Surendran
authors.forename: Peter L
authors.initials: PL
authors.lastname: Rady
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Quast
authors.forename: Gerald A
authors.initials: GA
authors.lastname: Campbell
authors.forename: Kimberlee M
authors.initials: KM
authors.lastname: Matalon
authors.forename: Stephen K
authors.initials: SK
authors.lastname: Tyring
authors.forename: Jingna
authors.initials: J
authors.lastname: Wei
authors.forename: Carmen S
authors.initials: CS
authors.lastname: Peden
authors.forename: E L
authors.initials: EL
authors.lastname: Ezell
authors.forename: Nicholas
authors.initials: N
authors.lastname: Muzyczka
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Mandel
mesh_terms: D000581:Amidohydrolases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D017825:Canavan Disease / Q000201:enzymology / Q000473:pathology / Q000628:therapy*; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D018345:Mice, Knockout; D010641:Phenotype; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D008164:Luminescent Proteins; D011994:Recombinant Proteins; D049452:Green Fluorescent Proteins; D000581:Amidohydrolases; C019751:aspartoacylase