title: Mitigation of cerebellar neuropathy in globoid cell leukodystrophy mice by AAV-mediated gene therapy.
abstract: Globoid cell leukodystrophy (GLD) is an autosomal recessive, lysosomal storage disease caused by deficiency of the enzyme galactocerebrosidase (GALC). The absence of GALC activity leads to the accumulation of the toxic substance psychosine and the preferential loss of myelinating cells in the central and peripheral nervous systems. Profound demyelination, astrogliosis and axonopathy are the hallmarks of the pathogenesis of GLD, and cerebellar ataxia is one of the dominant manifestations in adolescents and adults affected with GLD. To date, studies regarding cerebellar degeneration in GLD are limited. In this study, the efficacy of cerebellum-targeted gene therapy on the cerebellar neuropathology in twitcher mice (a murine model of GLD) has been validated. We observed degeneration of Purkinje cells, Bergmann glia, and granule cells in addition to astrocytosis and demyelination in the cerebellum of the twitcher mice. Ultrastructural analysis revealed dark cell degeneration and disintegration of the cellular composition of Purkinje cells in untreated twitcher mice. In addition, the expressions of neurotrophic factors CNTF, GDNF and IGF-I were up-regulated and the expression of BDNF was down-regulated. Intracerebellar-mediated gene therapy efficiently corrected enzymatic deficiency by direct transduction to Purkinje cells and cross-correction in other cell types in the cerebellum, leading to the amelioration of both neuroinflammation and demyelination. The population, dendritic territory, and axonal processes of Purkinje cells remained normal in the cerebellum of treated twitcher mice, where radial fibers of Bergmann glia spanned the molecular layer and collateral branches ensheathed the dendritic processes of Purkinje cells. Moreover, the aberrant expressions of neurotrophic factors were mitigated in the cerebellum of treated twitcher mice, indicating the preservation of cellular function in addition to maintaining the neuronal architecture. The life span of the treated twitcher mice was significantly prolonged and their neurobehavioral performance was improved. Taken together, our findings underscore the complexity of cerebellar neurodegeneration in GLD and highlight the potential effectiveness of gene therapy in mitigating neuropathological deficits in GLD and other neurodegenerative disorders in which Purkinje cells are involved.
authors.affiliation: Department of Pediatrics, Mackay Memorial Hospital, Taipei, Taiwan; Department of Medicine, Mackay Medical College, New Taipei, Taiwan; Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan. Electronic address: dslin@mmh.org.tw.
authors.forename: Dar-Shong
authors.initials: DS
authors.lastname: Lin
authors.affiliation: Department of Bioscience Technology, Chung Yuan Christian University, Chung-Li, Taiwan.
authors.forename: Chung-Der
authors.initials: CD
authors.lastname: Hsiao
authors.affiliation: National Institute of Cancer Research, National Health Research Institutes, Miaoli, Taiwan.
authors.forename: Allan Yueh-Luen
authors.initials: AY
authors.lastname: Lee
authors.affiliation: Department of Pediatrics, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Che-Sheng
authors.initials: CS
authors.lastname: Ho
authors.affiliation: Department of Chemical Engineering and Biotechnology, National Taipei University of Technology, Taipei, Taiwan.
authors.forename: Hsuan-Liang
authors.initials: HL
authors.lastname: Liu
authors.affiliation: Department of Laboratory Medicine, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Tuen-Jen
authors.initials: TJ
authors.lastname: Wang
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Yuan-Ren
authors.initials: YR
authors.lastname: Jian
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Jui-Cheng
authors.initials: JC
authors.lastname: Hsu
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Zon-Darr
authors.initials: ZD
authors.lastname: Huang
authors.affiliation: Department of Medical Research, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Tsung-Han
authors.initials: TH
authors.lastname: Lee
authors.affiliation: Department of Neurosurgery, Mackay Memorial Hospital, Taipei, Taiwan; Institute of Injury Prevention and Control, Taipei Medical University, Taipei, Taiwan. Electronic address: chiang66@gmail.com.
authors.forename: Ming-Fu
authors.initials: MF
authors.lastname: Chiang
mesh_terms: D000818:Animals; D019208:Brain-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D002526:Cerebellar Diseases / Q000235:genetics / Q000628:therapy*; D002531:Cerebellum / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D020934:Ciliary Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D005698:Galactosylceramidase / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D005911:Gliosis / Q000235:genetics / Q000378:metabolism; D007150:Immunohistochemistry; D007334:Insulin-Like Growth Factor I / Q000235:genetics / Q000378:metabolism; D053208:Kaplan-Meier Estimate; D007965:Leukodystrophy, Globoid Cell / Q000235:genetics / Q000628:therapy*; D008810:Mice, Inbred C57BL; D008818:Mice, Neurologic Mutants; D046529:Microscopy, Electron, Transmission; D009457:Neuroglia / Q000378:metabolism / Q000473:pathology; D011689:Purkinje Cells / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D019208:Brain-Derived Neurotrophic Factor; D020934:Ciliary Neurotrophic Factor; D051100:Glial Cell Line-Derived Neurotrophic Factor; D007334:Insulin-Like Growth Factor I; D005698:Galactosylceramidase
keywords: Adeno-associated virus; Globoid cell leukodystrophy; Lysosomal storage disease; Neurotrophic factor; Purkinje cells