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A Neuron-Specific Gene Therapy Relieves Motor Deficits in Pompe Disease Mice


In Pompe disease, deficient lysosomal acid α-glucosidase (GAA) activity causes glycogen accumulation in the muscles, which leads to weakness, cardiomyopathy, and respiratory failure. Although glycogen accumulation also occurs in the nervous system, the burden of neurological deficits in Pompe disease remains obscure. In this study, a neuron-specific gene therapy was administered to Pompe mice through intracerebroventricular injection of a viral vector carrying a neuron-specific promoter. The results revealed that gene therapy increased GAA activity and decreased glycogen content in the brain and spinal cord but not in the muscles of Pompe mice. Gene therapy only slightly increased the muscle strength of Pompe mice but substantially improved their performance on the rotarod, a test measuring motor coordination. Gene therapy also decreased astrogliosis and increased myelination in the brain and spinal cord of Pompe mice. Therefore, a neuron-specific treatment improved the motor coordination of Pompe mice by lowering glycogen accumulation, decreasing astrogliosis, and increasing myelination. These findings indicate that neurological deficits are responsible for a significant burden in Pompe disease.

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This work was funded by a grant from the Ministry of Science and Technology (103-2314-B-002-057-MY3) of Taiwan. The authors would like to thank the scientists from the Taiwan Mouse Clinic and the National Taiwan University Disease Animal Research Center, both of which are funded by the National Research Program for Biopharmaceuticals (NRPB), and doctor Kun-Ze Lee for the setting up of respiratory study. We thank Mika Ito and Naomi Takino (Jichi Medical University, Japan) for their help with the production of the AAV vectors.

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Authors and Affiliations



N.L, W.H., and Y.C. designed and conducted the study and performed statistical analysis. S.M. provided the AAVN vector. D.F., B.B., and L.T. supervised the immunohistochemistry and respiratory study. C.C, N.S., and K.C. performed the immunohistochemistry, functional study, and molecular studies. All authors participated in the manuscript preparation and approved the final version for submission.

Corresponding author

Correspondence to Yin-Hsiu Chien.

Ethics declarations

The authors have disclosed the potential conflicts of interest. The experimental procedures were approved and performed in accordance with the guidelines of the National Taiwan University College of Medicine and the College of Public Health Institutional Animal Care and Use Committee (IACUC No. 20120334). There is no human samples in this study and no inform consent is needed.

Conflict of Interest

S. M. owns equity in a gene therapy company (Gene Therapy Research Institution) that commercializes the use of AAV vectors for gene therapy applications. To the extent that the work in this manuscript increases the value of these commercial holdings, S. M. has a conflict of interest. No competing financial interests exist for other authors.

Electronic Supplementary Material

Supplementary Fig. 1
figure 8

(a) Distribution of vg. Real-time PCR analysis of vg copy number in the brain, spinal cord, liver, and quadriceps of the untreated (Pompe; red) and gene therapy-treated (Pompe-GT; blue) Pompe mice. The detection limit was 10 copies/μg DNA. * indicates p < 0.05. (b) Anti-GAA titers at the end of the study in the untreated, ERT-treated and gene therapy-treated Pompe mice. ERT, but not gene therapy, triggered a strong antibody response. * indicates p < 0.05. (JPEG 237 kb)

Supplementary Fig. 2
figure 9

Biodistribution of the injected virus. AAVN-GFP was injected in the same way as AAVN-GAA, and coronal sections were stained with anti-GFP. Images are at ×100 or ×400 magnification. Positive staining was observed in the neurons in the cortex (arrow; a and d) and Purkinje cells and other cell types in the cerebellum (arrow; b and e) but not in the striatum (c and f) or the anterior horn motor neurons in the cervical spinal cord (h and k). The light staining in the anterior horn motor neurons is likely background. Many fibers in the posterior horn stained positive (i and l). The window in panel L demonstrates a high magnification picture of the dot-like positive staining nerve fibers. (JPEG 1146 kb)

Supplementary Fig. 3
figure 10

Immunohistochemical staining of neuromuscular junctions in the diaphragm muscle. (a-d) Alexa Fluor 594-conjugated α-bungarotoxin staining of end plates of control mice (a) and untreated (Pompe; b), ERT-treated (Pompe-ERT; c), and gene therapy-treated (Pompe-GT; d) Pompe mice. End plates in the untreated Pompe mice were enlarged and fragmented (arrow), but neither ERT or gene therapy significantly improved the morphology of end plates. (e-h) α-bungarotoxin (red) and neurofilament plus ZNP-1 (green) triple staining in the untreated Pompe mice. (e) α-bungarotoxin staining. (f) Neurofilament plus ZNP-1 staining. (g) A merged view. (h) A high magnification picture of an end plate after triple staining. There was no significant abnormality in the neurofilaments plus ZNP-1. All images are at ×200 magnification except for panel h (×400). (JPEG 450 kb)

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Lee, NC., Hwu, WL., Muramatsu, SI. et al. A Neuron-Specific Gene Therapy Relieves Motor Deficits in Pompe Disease Mice. Mol Neurobiol 55, 5299–5309 (2018).

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  • Pompe disease
  • Glycogen
  • Gene therapy
  • Adeno-associated viral vector
  • Neuron-specific
  • Enzyme replacement therapy