Proteasome inhibitor (MG-132) treatment of mdx mice rescues the expression and membrane localization of dystrophin and dystrophin-associated proteins

Gloria Bonuccelli, Federica Sotgia, William Schubert, David S. Park, Philippe G. Frank, Scott E. Woodman, Luigi Insabato, Michael Cammer, Carlo Minetti, Michael P. Lisanti

Research output: Contribution to journalArticle

Abstract

Dystrophin, the protein product of the Duchenne muscular dystrophy (DMD) gene, is absent in the skeletal muscle of DMD patients and mdx mice. At the plasma membrane of skeletal muscle fibers, dystrophin associates with a multimeric protein complex, termed the dystrophin-glycoprotein complex (DGC). Protein members of this complex are normally absent or greatly reduced in dystrophin-deficient skeletal muscle fibers, and are thought to undergo degradation through an unknown pathway. As such, we reasoned that inhibition of the proteasomal degradation pathway might rescue the expression and subcellular localization of dystrophin-associated proteins. To test this hypothesis, we treated mdx with the well-characterized proteasomal inhibitor MG-132. First, we locally injected MG-132 into the gastrocnemius muscle, and observed the outcome after 24 hours. Next, we performed systemic treatment using an osmotic pump that allowed us to deliver different concentrations of the proteasomal inhibitor, over an 8-day period. By immunofluorescence and Western blot analysis, we show that administration of the proteasomal inhibitor MG-132 effectively rescues the expression levels and plasma membrane localization of dystrophin, β-dystroglycan, α-dystroglycan, and α-sarcoglycan in skeletal muscle fibers from mdx mice. Furthermore, we show that systemic treatment with the proteasomal inhibitor 1) reduces muscle membrane damage, as revealed by vital staining (with Evans blue dye) of the diaphragm and gastrocnemius muscle isolated from treated mdx mice, and 2) ameliorates the histopathological signs of muscular dystrophy, as judged by hematoxylin and eosin staining of muscle biopsies taken from treated mdx mice. Thus, the current study opens new and important avenues in our understanding of the pathogenesis of DMD. Most importantly, these new findings may have clinical implications for the pharmacological treatment of patients with DMD.

Original languageEnglish
Pages (from-to)1663-1675
Number of pages13
JournalAmerican Journal of Pathology
Volume163
Issue number4
Publication statusPublished - Oct 1 2003

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Dystrophin-Associated Proteins
Inbred mdx Mouse
Dystrophin
Proteasome Inhibitors
Duchenne Muscular Dystrophy
Skeletal Muscle Fibers
Membranes
Dystroglycans
Skeletal Muscle
Sarcoglycans
Cell Membrane
Staining and Labeling
Therapeutics
Muscles
Evans Blue
Proteins
Muscular Dystrophies
Hematoxylin
Eosine Yellowish-(YS)
Diaphragm

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Bonuccelli, G., Sotgia, F., Schubert, W., Park, D. S., Frank, P. G., Woodman, S. E., ... Lisanti, M. P. (2003). Proteasome inhibitor (MG-132) treatment of mdx mice rescues the expression and membrane localization of dystrophin and dystrophin-associated proteins. American Journal of Pathology, 163(4), 1663-1675.

Proteasome inhibitor (MG-132) treatment of mdx mice rescues the expression and membrane localization of dystrophin and dystrophin-associated proteins. / Bonuccelli, Gloria; Sotgia, Federica; Schubert, William; Park, David S.; Frank, Philippe G.; Woodman, Scott E.; Insabato, Luigi; Cammer, Michael; Minetti, Carlo; Lisanti, Michael P.

In: American Journal of Pathology, Vol. 163, No. 4, 01.10.2003, p. 1663-1675.

Research output: Contribution to journalArticle

Bonuccelli, G, Sotgia, F, Schubert, W, Park, DS, Frank, PG, Woodman, SE, Insabato, L, Cammer, M, Minetti, C & Lisanti, MP 2003, 'Proteasome inhibitor (MG-132) treatment of mdx mice rescues the expression and membrane localization of dystrophin and dystrophin-associated proteins', American Journal of Pathology, vol. 163, no. 4, pp. 1663-1675.
Bonuccelli, Gloria ; Sotgia, Federica ; Schubert, William ; Park, David S. ; Frank, Philippe G. ; Woodman, Scott E. ; Insabato, Luigi ; Cammer, Michael ; Minetti, Carlo ; Lisanti, Michael P. / Proteasome inhibitor (MG-132) treatment of mdx mice rescues the expression and membrane localization of dystrophin and dystrophin-associated proteins. In: American Journal of Pathology. 2003 ; Vol. 163, No. 4. pp. 1663-1675.
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abstract = "Dystrophin, the protein product of the Duchenne muscular dystrophy (DMD) gene, is absent in the skeletal muscle of DMD patients and mdx mice. At the plasma membrane of skeletal muscle fibers, dystrophin associates with a multimeric protein complex, termed the dystrophin-glycoprotein complex (DGC). Protein members of this complex are normally absent or greatly reduced in dystrophin-deficient skeletal muscle fibers, and are thought to undergo degradation through an unknown pathway. As such, we reasoned that inhibition of the proteasomal degradation pathway might rescue the expression and subcellular localization of dystrophin-associated proteins. To test this hypothesis, we treated mdx with the well-characterized proteasomal inhibitor MG-132. First, we locally injected MG-132 into the gastrocnemius muscle, and observed the outcome after 24 hours. Next, we performed systemic treatment using an osmotic pump that allowed us to deliver different concentrations of the proteasomal inhibitor, over an 8-day period. By immunofluorescence and Western blot analysis, we show that administration of the proteasomal inhibitor MG-132 effectively rescues the expression levels and plasma membrane localization of dystrophin, β-dystroglycan, α-dystroglycan, and α-sarcoglycan in skeletal muscle fibers from mdx mice. Furthermore, we show that systemic treatment with the proteasomal inhibitor 1) reduces muscle membrane damage, as revealed by vital staining (with Evans blue dye) of the diaphragm and gastrocnemius muscle isolated from treated mdx mice, and 2) ameliorates the histopathological signs of muscular dystrophy, as judged by hematoxylin and eosin staining of muscle biopsies taken from treated mdx mice. Thus, the current study opens new and important avenues in our understanding of the pathogenesis of DMD. Most importantly, these new findings may have clinical implications for the pharmacological treatment of patients with DMD.",
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AU - Park, David S.

AU - Frank, Philippe G.

AU - Woodman, Scott E.

AU - Insabato, Luigi

AU - Cammer, Michael

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