Prolonged expression of a lysosomal enzyme in mouse liver after Sleeping Beauty transposon-mediated gene delivery: Implications for non-viral gene therapy of mucopolysaccharidoses

Elena L Aronovich, Jason B. Bell, Lalitha Belur, Roland Gunther, Brenda Koniar, David C C Erickson, Patricia A. Schachern, Ilze Matise, R. Scott Mclvor, Chester B Whitley, Perry B Hackett

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Abstract

Background: The Sleeping Beauty (SB) transposon system is a non-viral vector system that can integrate precise sequences into chromosomes. We evaluated the SB transposon system as a tool for gene therapy of mucopolysaccharidosis (MPS) types I and VII. Methods: We constructed SB transposon plasmids for high-level expression of human β-glucuronidase (hGUSB) or α-L-iduronidase (hIDUA). Plasmids were delivered with and without SB transposase to mouse liver by rapid, high-volume tail-vein injection. We studied the duration of expressed therapeutic enzyme activity, transgene presence by PCR, lysosomal pathology by toluidine blue staining and cell-mediated immune response histologically and by immunohistochemical staining. Results: Transgene frequency, distribution of transgene and enzyme expression in liver and the level of transgenic enzyme required for amelioration of lysosomal pathology were estimated in MPS I and VII mice. Without immunomodulation, initial GUSB and IDUA activities in plasma reached >100-fold of wild-type (WT) levels but fell to background within 4 weeks post-injection. In immunomodulated transposon-treated MPS I mice plasma IDUA persisted for over 3 months at up to 100-fold WT activity in one-third of MPS I mice, which was sufficient to reverse lysosomal pathology in the liver and, partially, in distant organs. Histological and immunohistochemical examination of liver sections in IDUA transposon-treated WT mice revealed inflammation 10 days post-injection consisting predominantly of mononuclear cells, some of which were CD4- or CD8-positive. Conclusions: Our results demonstrate the feasibility of achieving prolonged expression of lysosomal enzymes in the liver and reversing MPS disease in adult mice with a single dose of therapeutic SB transposons.

Original languageEnglish (US)
Pages (from-to)403-415
Number of pages13
JournalJournal of Gene Medicine
Volume9
Issue number5
DOIs
StatePublished - May 1 2007

Fingerprint

Mucopolysaccharidoses
Beauty
Genetic Therapy
Mucopolysaccharidosis I
Liver
Mucopolysaccharidosis VII
Enzymes
Transgenes
Genes
Pathology
Injections
Plasmids
Iduronidase
Staining and Labeling
Transposases
Tolonium Chloride
Immunomodulation
Glucuronidase
Tail
Veins

Keywords

  • Animal model
  • Immune response
  • Inherited disease
  • α-L-iduronidase
  • β-glucuronidase

Cite this

Prolonged expression of a lysosomal enzyme in mouse liver after Sleeping Beauty transposon-mediated gene delivery : Implications for non-viral gene therapy of mucopolysaccharidoses. / Aronovich, Elena L; Bell, Jason B.; Belur, Lalitha; Gunther, Roland; Koniar, Brenda; Erickson, David C C; Schachern, Patricia A.; Matise, Ilze; Mclvor, R. Scott; Whitley, Chester B; Hackett, Perry B.

In: Journal of Gene Medicine, Vol. 9, No. 5, 01.05.2007, p. 403-415.

Research output: Contribution to journalArticle

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abstract = "Background: The Sleeping Beauty (SB) transposon system is a non-viral vector system that can integrate precise sequences into chromosomes. We evaluated the SB transposon system as a tool for gene therapy of mucopolysaccharidosis (MPS) types I and VII. Methods: We constructed SB transposon plasmids for high-level expression of human β-glucuronidase (hGUSB) or α-L-iduronidase (hIDUA). Plasmids were delivered with and without SB transposase to mouse liver by rapid, high-volume tail-vein injection. We studied the duration of expressed therapeutic enzyme activity, transgene presence by PCR, lysosomal pathology by toluidine blue staining and cell-mediated immune response histologically and by immunohistochemical staining. Results: Transgene frequency, distribution of transgene and enzyme expression in liver and the level of transgenic enzyme required for amelioration of lysosomal pathology were estimated in MPS I and VII mice. Without immunomodulation, initial GUSB and IDUA activities in plasma reached >100-fold of wild-type (WT) levels but fell to background within 4 weeks post-injection. In immunomodulated transposon-treated MPS I mice plasma IDUA persisted for over 3 months at up to 100-fold WT activity in one-third of MPS I mice, which was sufficient to reverse lysosomal pathology in the liver and, partially, in distant organs. Histological and immunohistochemical examination of liver sections in IDUA transposon-treated WT mice revealed inflammation 10 days post-injection consisting predominantly of mononuclear cells, some of which were CD4- or CD8-positive. Conclusions: Our results demonstrate the feasibility of achieving prolonged expression of lysosomal enzymes in the liver and reversing MPS disease in adult mice with a single dose of therapeutic SB transposons.",
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T1 - Prolonged expression of a lysosomal enzyme in mouse liver after Sleeping Beauty transposon-mediated gene delivery

T2 - Implications for non-viral gene therapy of mucopolysaccharidoses

AU - Aronovich, Elena L

AU - Bell, Jason B.

AU - Belur, Lalitha

AU - Gunther, Roland

AU - Koniar, Brenda

AU - Erickson, David C C

AU - Schachern, Patricia A.

AU - Matise, Ilze

AU - Mclvor, R. Scott

AU - Whitley, Chester B

AU - Hackett, Perry B

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AB - Background: The Sleeping Beauty (SB) transposon system is a non-viral vector system that can integrate precise sequences into chromosomes. We evaluated the SB transposon system as a tool for gene therapy of mucopolysaccharidosis (MPS) types I and VII. Methods: We constructed SB transposon plasmids for high-level expression of human β-glucuronidase (hGUSB) or α-L-iduronidase (hIDUA). Plasmids were delivered with and without SB transposase to mouse liver by rapid, high-volume tail-vein injection. We studied the duration of expressed therapeutic enzyme activity, transgene presence by PCR, lysosomal pathology by toluidine blue staining and cell-mediated immune response histologically and by immunohistochemical staining. Results: Transgene frequency, distribution of transgene and enzyme expression in liver and the level of transgenic enzyme required for amelioration of lysosomal pathology were estimated in MPS I and VII mice. Without immunomodulation, initial GUSB and IDUA activities in plasma reached >100-fold of wild-type (WT) levels but fell to background within 4 weeks post-injection. In immunomodulated transposon-treated MPS I mice plasma IDUA persisted for over 3 months at up to 100-fold WT activity in one-third of MPS I mice, which was sufficient to reverse lysosomal pathology in the liver and, partially, in distant organs. Histological and immunohistochemical examination of liver sections in IDUA transposon-treated WT mice revealed inflammation 10 days post-injection consisting predominantly of mononuclear cells, some of which were CD4- or CD8-positive. Conclusions: Our results demonstrate the feasibility of achieving prolonged expression of lysosomal enzymes in the liver and reversing MPS disease in adult mice with a single dose of therapeutic SB transposons.

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