Functional substitution by TAT-utrophin in dystrophin-deficient mice

Kevin J. Sonnemann, Hanke Heun-Johnson, Amy J. Turner, Kristen A. Baltgalvis, Dawn A. Lowe, James M. Ervasti

Research output: Contribution to journalArticle

63 Citations (Scopus)

Abstract

Background: The loss of dystrophin compromises muscle cell membrane stability and causes Duchenne muscular dystrophy and/or various forms of cardiomyopathy. Increased expression of the dystrophin homolog utrophin by gene delivery or pharmacologic up-regulation has been demonstrated to restore membrane integrity and improve the phenotype in the dystrophin-deficient mdx mouse. However, the lack of a viable therapy in humans predicates the need to explore alternative methods to combat dystrophin deficiency. We investigated whether systemic administration of recombinant full-length utrophin (Utr) or DR4-21 "micro" utrophin (mUtr) protein modified with the cell-penetrating TAT protein transduction domain could attenuate the phenotype of mdx mice. Methods and Findings: Recombinant TAT-Utr and TAT-μUtr proteins were expressed using the baculovirus system and purified using FLAG-affinity chromatography. Age-matched mdx mice received six twice-weekly intraperitoneal injections of either recombinant protein or PBS. Three days after the final injection, mice were analyzed for several phenotypic parameters of dystrophin deficiency. Injected TAT-μmUtr transduced all tissues examined, integrated with members of the dystrophin complex, reduced serum levels of creatine kinase (11,290±920 U versus 5,950±1,120 U; PBS versus TAT), the prevalence of muscle degeneration/regeneration (54%±5% versus 37%±4% of centrally nucleated fibers; PBS versus TAT), the susceptibility to eccentric contraction-induced force drop (72%±5% versus 40%±8% drop; PBS versus TAT), and increased specific force production (9.7±1.1 N/cm2 versus 12.8±0.9 N/cm2; PBS versus TAT). Conclusions: These results are, to our knowledge, the first to establish the efficacy and feasibility of TAT-utrophin-based constructs as a novel direct protein-replacement therapy for the treatment of skeletal and cardiac muscle diseases caused by loss of dystrophin.

Original languageEnglish (US)
Article numbere1000083
JournalPLoS medicine
Volume6
Issue number5
DOIs
StatePublished - May 1 2009

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Utrophin
Dystrophin
Inbred mdx Mouse
Phenotype
Proteins
Duchenne Muscular Dystrophy
Baculoviridae
Creatine Kinase
Intraperitoneal Injections
Cardiomyopathies
Affinity Chromatography
Recombinant Proteins
Muscle Cells
Regeneration
Heart Diseases
Myocardium
Skeletal Muscle
Up-Regulation
Therapeutics
Cell Membrane

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Functional substitution by TAT-utrophin in dystrophin-deficient mice. / Sonnemann, Kevin J.; Heun-Johnson, Hanke; Turner, Amy J.; Baltgalvis, Kristen A.; Lowe, Dawn A.; Ervasti, James M.

In: PLoS medicine, Vol. 6, No. 5, e1000083, 01.05.2009.

Research output: Contribution to journalArticle

Sonnemann, Kevin J. ; Heun-Johnson, Hanke ; Turner, Amy J. ; Baltgalvis, Kristen A. ; Lowe, Dawn A. ; Ervasti, James M. / Functional substitution by TAT-utrophin in dystrophin-deficient mice. In: PLoS medicine. 2009 ; Vol. 6, No. 5.
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AU - Heun-Johnson, Hanke

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