Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of duchenne muscular dystrophy

Yadong Zhang, Yongping Yue, Liang Li, Chady H. Hakim, Keqing Zhang, Gail Thomas, Dongsheng Duan

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Neuronal nitric oxide synthase (nNOS) membrane delocalization contributes to the pathogenesis of Duchenne muscular dystrophy (DMD) by promoting functional muscle ischemia and exacerbating muscle injury during exercise. We have previously shown that supra-physiological expression of nNOS-binding mini-dystrophin restores normal blood flow regulation and prevents functional ischemia in transgenic mdx mice, a DMD model. A critical next issue is whether systemic dual adeno-associated virus (AAV) gene therapy can restore nNOS-binding mini-dystrophin expression and mitigate muscle activity-related functional ischemia and injury. Here, we performed systemic gene transfer in mdx and mdx4cv mice using a pair of dual AAV vectors that expressed a 6 kb nNOS-binding mini-dystrophin gene. Vectors were packaged in tyrosine mutant AAV-9 and co-injected (5 × 1012 viral genome particles/vector/mouse) via the tail vein to 1-month-old dystrophin-null mice. Four months later, we observed 30-50% mini-dystrophin positive myofibers in limb muscles. Treatment ameliorated histopathology, increased muscle force and protected against eccentric contractioninduced injury. Importantly, dual AAV therapy successfully prevented chronic exercise-induced muscle force drop. Doppler hemodynamic assay further showed that therapy attenuated adrenergic vasoconstriction in contracting muscle. Our results suggest that partial transduction can still ameliorate nNOS delocalization-associated functional deficiency. Further evaluation of nNOS binding mini-dystrophin dual AAV vectors is warranted in dystrophic dogs and eventually in human patients.

Original languageEnglish (US)
Article numberddt224
Pages (from-to)3720-3729
Number of pages10
JournalHuman Molecular Genetics
Volume22
Issue number18
DOIs
StatePublished - Sep 1 2013

Fingerprint

Exercise Therapy
Dependovirus
Duchenne Muscular Dystrophy
Nitric Oxide Synthase Type I
Dystrophin
Ischemia
Muscles
Wounds and Injuries
Inbred mdx Mouse
Exercise
Viral Genome
Vasoconstriction
Genetic Therapy
Virion
Adrenergic Agents
Transgenic Mice
Genes
Tyrosine
Tail
Veins

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Genetics
  • Genetics(clinical)

Cite this

Zhang, Yadong ; Yue, Yongping ; Li, Liang ; Hakim, Chady H. ; Zhang, Keqing ; Thomas, Gail ; Duan, Dongsheng. / Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of duchenne muscular dystrophy. In: Human Molecular Genetics. 2013 ; Vol. 22, No. 18. pp. 3720-3729.
@article{20101d8c883a43f79c83f51913c7ef15,
title = "Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of duchenne muscular dystrophy",
abstract = "Neuronal nitric oxide synthase (nNOS) membrane delocalization contributes to the pathogenesis of Duchenne muscular dystrophy (DMD) by promoting functional muscle ischemia and exacerbating muscle injury during exercise. We have previously shown that supra-physiological expression of nNOS-binding mini-dystrophin restores normal blood flow regulation and prevents functional ischemia in transgenic mdx mice, a DMD model. A critical next issue is whether systemic dual adeno-associated virus (AAV) gene therapy can restore nNOS-binding mini-dystrophin expression and mitigate muscle activity-related functional ischemia and injury. Here, we performed systemic gene transfer in mdx and mdx4cv mice using a pair of dual AAV vectors that expressed a 6 kb nNOS-binding mini-dystrophin gene. Vectors were packaged in tyrosine mutant AAV-9 and co-injected (5 × 1012 viral genome particles/vector/mouse) via the tail vein to 1-month-old dystrophin-null mice. Four months later, we observed 30-50{\%} mini-dystrophin positive myofibers in limb muscles. Treatment ameliorated histopathology, increased muscle force and protected against eccentric contractioninduced injury. Importantly, dual AAV therapy successfully prevented chronic exercise-induced muscle force drop. Doppler hemodynamic assay further showed that therapy attenuated adrenergic vasoconstriction in contracting muscle. Our results suggest that partial transduction can still ameliorate nNOS delocalization-associated functional deficiency. Further evaluation of nNOS binding mini-dystrophin dual AAV vectors is warranted in dystrophic dogs and eventually in human patients.",
author = "Yadong Zhang and Yongping Yue and Liang Li and Hakim, {Chady H.} and Keqing Zhang and Gail Thomas and Dongsheng Duan",
year = "2013",
month = "9",
day = "1",
doi = "10.1093/hmg/ddt224",
language = "English (US)",
volume = "22",
pages = "3720--3729",
journal = "Human Molecular Genetics",
issn = "0964-6906",
publisher = "Oxford University Press",
number = "18",

}

Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of duchenne muscular dystrophy. / Zhang, Yadong; Yue, Yongping; Li, Liang; Hakim, Chady H.; Zhang, Keqing; Thomas, Gail; Duan, Dongsheng.

In: Human Molecular Genetics, Vol. 22, No. 18, ddt224, 01.09.2013, p. 3720-3729.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dual AAV therapy ameliorates exercise-induced muscle injury and functional ischemia in murine models of duchenne muscular dystrophy

AU - Zhang, Yadong

AU - Yue, Yongping

AU - Li, Liang

AU - Hakim, Chady H.

AU - Zhang, Keqing

AU - Thomas, Gail

AU - Duan, Dongsheng

PY - 2013/9/1

Y1 - 2013/9/1

N2 - Neuronal nitric oxide synthase (nNOS) membrane delocalization contributes to the pathogenesis of Duchenne muscular dystrophy (DMD) by promoting functional muscle ischemia and exacerbating muscle injury during exercise. We have previously shown that supra-physiological expression of nNOS-binding mini-dystrophin restores normal blood flow regulation and prevents functional ischemia in transgenic mdx mice, a DMD model. A critical next issue is whether systemic dual adeno-associated virus (AAV) gene therapy can restore nNOS-binding mini-dystrophin expression and mitigate muscle activity-related functional ischemia and injury. Here, we performed systemic gene transfer in mdx and mdx4cv mice using a pair of dual AAV vectors that expressed a 6 kb nNOS-binding mini-dystrophin gene. Vectors were packaged in tyrosine mutant AAV-9 and co-injected (5 × 1012 viral genome particles/vector/mouse) via the tail vein to 1-month-old dystrophin-null mice. Four months later, we observed 30-50% mini-dystrophin positive myofibers in limb muscles. Treatment ameliorated histopathology, increased muscle force and protected against eccentric contractioninduced injury. Importantly, dual AAV therapy successfully prevented chronic exercise-induced muscle force drop. Doppler hemodynamic assay further showed that therapy attenuated adrenergic vasoconstriction in contracting muscle. Our results suggest that partial transduction can still ameliorate nNOS delocalization-associated functional deficiency. Further evaluation of nNOS binding mini-dystrophin dual AAV vectors is warranted in dystrophic dogs and eventually in human patients.

AB - Neuronal nitric oxide synthase (nNOS) membrane delocalization contributes to the pathogenesis of Duchenne muscular dystrophy (DMD) by promoting functional muscle ischemia and exacerbating muscle injury during exercise. We have previously shown that supra-physiological expression of nNOS-binding mini-dystrophin restores normal blood flow regulation and prevents functional ischemia in transgenic mdx mice, a DMD model. A critical next issue is whether systemic dual adeno-associated virus (AAV) gene therapy can restore nNOS-binding mini-dystrophin expression and mitigate muscle activity-related functional ischemia and injury. Here, we performed systemic gene transfer in mdx and mdx4cv mice using a pair of dual AAV vectors that expressed a 6 kb nNOS-binding mini-dystrophin gene. Vectors were packaged in tyrosine mutant AAV-9 and co-injected (5 × 1012 viral genome particles/vector/mouse) via the tail vein to 1-month-old dystrophin-null mice. Four months later, we observed 30-50% mini-dystrophin positive myofibers in limb muscles. Treatment ameliorated histopathology, increased muscle force and protected against eccentric contractioninduced injury. Importantly, dual AAV therapy successfully prevented chronic exercise-induced muscle force drop. Doppler hemodynamic assay further showed that therapy attenuated adrenergic vasoconstriction in contracting muscle. Our results suggest that partial transduction can still ameliorate nNOS delocalization-associated functional deficiency. Further evaluation of nNOS binding mini-dystrophin dual AAV vectors is warranted in dystrophic dogs and eventually in human patients.

UR - http://www.scopus.com/inward/record.url?scp=84882936501&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84882936501&partnerID=8YFLogxK

U2 - 10.1093/hmg/ddt224

DO - 10.1093/hmg/ddt224

M3 - Article

VL - 22

SP - 3720

EP - 3729

JO - Human Molecular Genetics

JF - Human Molecular Genetics

SN - 0964-6906

IS - 18

M1 - ddt224

ER -