Prolonged Expression of Secreted Enzymes in Dogs after Liver-Directed Delivery of Sleeping Beauty Transposons

Implications for Non-Viral Gene Therapy of Systemic Disease

Elena L. Aronovich, Kendra A. Hyland, Bryan C. Hall, Jason B. Bell, Erik R. Olson, Myra Urness Rusten, David W. Hunter, N. Matthew Ellinwood, R. Scott McIvor, Perry B. Hackett

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The non-viral, integrating Sleeping Beauty (SB) transposon system is efficient in treating systemic monogenic disease in mice, including hemophilia A and B caused by deficiency of blood clotting factors and mucopolysaccharidosis types I and VII caused by α-L-iduronidase (IDUA) and β-glucuronidase (GUSB) deficiency, respectively. Modified approaches of the hydrodynamics-based procedure to deliver transposons to the liver in dogs were recently reported. Using the transgenic canine reporter secreted alkaline phosphatase (cSEAP), transgenic protein in the plasma was demonstrated for up to 6 weeks post infusion. This study reports that immunosuppression of dogs with gadolinium chloride (GdCl3) prolonged the presence of cSEAP in the circulation up to 5.5 months after a single vector infusion. Transgene expression declined gradually but appeared to stabilize after about 2 months at approximately fourfold baseline level. Durability of transgenic protein expression in the plasma was inversely associated with transient increase of liver enzymes alanine transaminase and aspartate transaminase in response to the plasmid delivery procedure, which suggests a deleterious effect of hepatocellular toxicity on transgene expression. GdCl3 treatment was ineffective for repeat vector infusions. In parallel studies, dogs were infused with potentially therapeutic transposons. Activities of transgenic IDUA and GUSB in plasma peaked at 50-350% of wildtype, but in the absence of immunosuppression lasted only a few days. Transposition was detectable by excision assay only when the most efficient transposase, SB100X, was used. Dogs infused with transposons encoding canine clotting factor IX (cFIX) were treated with GdCl3 and showed expression profiles similar to those in cSEAP-infused dogs, with expression peaking at 40% wt (2 μg/mL). It is concluded that GdCl3 can support extended transgene expression after hydrodynamic introduction of SB transposons in dogs, but that alternative regimens will be required to achieve therapeutic levels of transgene products.

Original languageEnglish (US)
Pages (from-to)551-564
Number of pages14
JournalHuman gene therapy
Volume28
Issue number7
DOIs
StatePublished - Jul 1 2017

Fingerprint

Beauty
Genetic Therapy
Dogs
Transgenes
Canidae
Liver
Mucopolysaccharidosis VII
Enzymes
Iduronidase
Alkaline Phosphatase
Blood Coagulation Factors
Hydrodynamics
Immunosuppression
Mucopolysaccharidosis I
Transposases
Hemophilia B
Factor VII
Factor IX
Glucuronidase
Hemophilia A

Keywords

  • Monogenic disease
  • hemophilia
  • hydrodynamicsbased delivery
  • immune response
  • mucopolysaccharidosis

Cite this

Prolonged Expression of Secreted Enzymes in Dogs after Liver-Directed Delivery of Sleeping Beauty Transposons : Implications for Non-Viral Gene Therapy of Systemic Disease. / Aronovich, Elena L.; Hyland, Kendra A.; Hall, Bryan C.; Bell, Jason B.; Olson, Erik R.; Rusten, Myra Urness; Hunter, David W.; Matthew Ellinwood, N.; Scott McIvor, R.; Hackett, Perry B.

In: Human gene therapy, Vol. 28, No. 7, 01.07.2017, p. 551-564.

Research output: Contribution to journalArticle

Aronovich, Elena L. ; Hyland, Kendra A. ; Hall, Bryan C. ; Bell, Jason B. ; Olson, Erik R. ; Rusten, Myra Urness ; Hunter, David W. ; Matthew Ellinwood, N. ; Scott McIvor, R. ; Hackett, Perry B. / Prolonged Expression of Secreted Enzymes in Dogs after Liver-Directed Delivery of Sleeping Beauty Transposons : Implications for Non-Viral Gene Therapy of Systemic Disease. In: Human gene therapy. 2017 ; Vol. 28, No. 7. pp. 551-564.
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abstract = "The non-viral, integrating Sleeping Beauty (SB) transposon system is efficient in treating systemic monogenic disease in mice, including hemophilia A and B caused by deficiency of blood clotting factors and mucopolysaccharidosis types I and VII caused by α-L-iduronidase (IDUA) and β-glucuronidase (GUSB) deficiency, respectively. Modified approaches of the hydrodynamics-based procedure to deliver transposons to the liver in dogs were recently reported. Using the transgenic canine reporter secreted alkaline phosphatase (cSEAP), transgenic protein in the plasma was demonstrated for up to 6 weeks post infusion. This study reports that immunosuppression of dogs with gadolinium chloride (GdCl3) prolonged the presence of cSEAP in the circulation up to 5.5 months after a single vector infusion. Transgene expression declined gradually but appeared to stabilize after about 2 months at approximately fourfold baseline level. Durability of transgenic protein expression in the plasma was inversely associated with transient increase of liver enzymes alanine transaminase and aspartate transaminase in response to the plasmid delivery procedure, which suggests a deleterious effect of hepatocellular toxicity on transgene expression. GdCl3 treatment was ineffective for repeat vector infusions. In parallel studies, dogs were infused with potentially therapeutic transposons. Activities of transgenic IDUA and GUSB in plasma peaked at 50-350{\%} of wildtype, but in the absence of immunosuppression lasted only a few days. Transposition was detectable by excision assay only when the most efficient transposase, SB100X, was used. Dogs infused with transposons encoding canine clotting factor IX (cFIX) were treated with GdCl3 and showed expression profiles similar to those in cSEAP-infused dogs, with expression peaking at 40{\%} wt (2 μg/mL). It is concluded that GdCl3 can support extended transgene expression after hydrodynamic introduction of SB transposons in dogs, but that alternative regimens will be required to achieve therapeutic levels of transgene products.",
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