Novel Vector Design and Hexosaminidase Variant Enabling Self-Complementary Adeno-Associated Virus for the Treatment of Tay-Sachs Disease

Subha Karumuthil-Melethil, Sahana Nagabhushan Kalburgi, Patrick Thompson, Michael Tropak, Michael D. Kaytor, John G. Keimel, Brian L. Mark, Don Mahuran, Jagdeep S. Walia, Steven J. Gray

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

GM2 gangliosidosis is a family of three genetic neurodegenerative disorders caused by the accumulation of GM2 ganglioside (GM2) in neuronal tissue. Two of these are due to the deficiency of the heterodimeric (α-β), "A" isoenzyme of lysosomal β-hexosaminidase (HexA). Mutations in the α-subunit (encoded by HEXA) lead to Tay-Sachs disease (TSD), whereas mutations in the β-subunit (encoded by HEXB) lead to Sandhoff disease (SD). The third form results from a deficiency of the GM2 activator protein (GM2AP), a substrate-specific cofactor for HexA. In their infantile, acute forms, these diseases rapidly progress with mental and psychomotor deterioration resulting in death by approximately 4 years of age. After gene transfer that overexpresses one of the deficient subunits, the amount of HexA heterodimer formed would empirically be limited by the availability of the other endogenous Hex subunit. The present study used a new variant of the human HexA α-subunit, μ, incorporating critical sequences from the β-subunit that produce a stable homodimer (HexM) and promote functional interactions with the GM2AP-GM2 complex. We report the design of a compact adeno-associated viral (AAV) genome using a synthetic promoter-intron combination to allow self-complementary (sc) packaging of the HEXM gene. Also, a previously published capsid mutant, AAV9.47, was used to deliver the gene to brain and spinal cord while having restricted biodistribution to the liver. The novel capsid and cassette design combination was characterized in vivo in TSD mice for its ability to efficiently transduce cells in the central nervous system when delivered intravenously in both adult and neonatal mice. This study demonstrates that the modified HexM is capable of degrading long-standing GM2 storage in mice, and it further demonstrates the potential of this novel scAAV vector design to facilitate widespread distribution of the HEXM gene or potentially other similar-sized genes to the nervous system.

Original languageEnglish (US)
Pages (from-to)509-521
Number of pages13
JournalHuman Gene Therapy
Volume27
Issue number7
DOIs
StatePublished - Jul 1 2016

Fingerprint

Tay-Sachs Disease
Hexosaminidases
Dependovirus
G(M2) Activator Protein
Genes
Capsid
GM2 Gangliosidosis
Sandhoff Disease
G(M2) Ganglioside
Mutation
Inborn Genetic Diseases
Aptitude
Viral Genome
Acute Disease
Product Packaging
Neurodegenerative Diseases
Introns
Nervous System
Isoenzymes
Spinal Cord

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Novel Vector Design and Hexosaminidase Variant Enabling Self-Complementary Adeno-Associated Virus for the Treatment of Tay-Sachs Disease. / Karumuthil-Melethil, Subha; Nagabhushan Kalburgi, Sahana; Thompson, Patrick; Tropak, Michael; Kaytor, Michael D.; Keimel, John G.; Mark, Brian L.; Mahuran, Don; Walia, Jagdeep S.; Gray, Steven J.

In: Human Gene Therapy, Vol. 27, No. 7, 01.07.2016, p. 509-521.

Research output: Contribution to journalArticle

Karumuthil-Melethil, S, Nagabhushan Kalburgi, S, Thompson, P, Tropak, M, Kaytor, MD, Keimel, JG, Mark, BL, Mahuran, D, Walia, JS & Gray, SJ 2016, 'Novel Vector Design and Hexosaminidase Variant Enabling Self-Complementary Adeno-Associated Virus for the Treatment of Tay-Sachs Disease', Human Gene Therapy, vol. 27, no. 7, pp. 509-521. https://doi.org/10.1089/hum.2016.013
Karumuthil-Melethil S, Nagabhushan Kalburgi S, Thompson P, Tropak M, Kaytor MD, Keimel JG et al. Novel Vector Design and Hexosaminidase Variant Enabling Self-Complementary Adeno-Associated Virus for the Treatment of Tay-Sachs Disease. Human Gene Therapy. 2016 Jul 1;27(7):509-521. https://doi.org/10.1089/hum.2016.013
Karumuthil-Melethil, Subha ; Nagabhushan Kalburgi, Sahana ; Thompson, Patrick ; Tropak, Michael ; Kaytor, Michael D. ; Keimel, John G. ; Mark, Brian L. ; Mahuran, Don ; Walia, Jagdeep S. ; Gray, Steven J. / Novel Vector Design and Hexosaminidase Variant Enabling Self-Complementary Adeno-Associated Virus for the Treatment of Tay-Sachs Disease. In: Human Gene Therapy. 2016 ; Vol. 27, No. 7. pp. 509-521.
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