Recombinant MG53 Protein Modulates Therapeutic Cell Membrane Repair in Treatment of Muscular Dystrophy
Science Translational Medicine • 20 Jun 2012 • Vol 4, Issue 139 • p. 139ra85 • DOI: 10.1126/scitranslmed.3003921
Mending Muscle
To repair a torn muscle, one might require a little bit of ice and a lot of rest. For those with Duchenne muscular dystrophy (DMD), however, muscle degeneration is not as easily repaired, and patients ultimately experience difficulty standing, walking, and breathing. DMD results from a lack of the protein dystrophin, which is located at the cell membrane to help muscle fibers repair themselves. There is no cure for DMD, but Weisleder and colleagues have now shown that exogenous delivery of a different repair protein, Mitsugumin 53 (MG53), to cells can prevent muscle damage in cell culture and in mice.
The authors first showed that muscle and nonmuscle cells treated with recombinant human MG53 (rhMG53) in vitro were resistant to mechanical, chemical, and photo damage because MG53 localized to the injury site and provided protection. In vivo, Weisleder and colleagues showed that dystrophin-deficient mdx mice treated intramuscularly or intravenously with rhMG53 displayed reduced muscle damage and decreased muscle pathology compared to saline-treated controls, even in the presence of a membrane-damaging toxin. This repair process also worked in muscle fibers isolated from mdx mice that were deficient in either of two natural repair proteins, MG53 or dysferlin, suggesting that exogenous delivery of rhMG53 works by a new mechanism—other than the intracellular machinery—to patch up damaged cell membrane.
Soluble MG53 protein therapy could be a viable treatment for DMD that avoids the well-known limitations of dystrophin gene replacement therapy. Toward translation, Weisleder et al. have further demonstrated that exogenous MG53 is nontoxic and safe in animals. The ability of the protein to preserve muscle function and to enhance repair capacity in humans has yet to be shown, but additional studies in larger animals and human muscle fibers will give a clearer indication of its therapeutic potential.
Abstract
Mitsugumin 53 (MG53), a muscle-specific TRIM family protein, is an essential component of the cell membrane repair machinery. Here, we examined the translational value of targeting MG53 function in tissue repair and regenerative medicine. Although native MG53 protein is principally restricted to skeletal and cardiac muscle tissues, beneficial effects that protect against cellular injuries are present in nonmuscle cells with overexpression of MG53. In addition to the intracellular action of MG53, injury to the cell membrane exposes a signal that can be detected by MG53, allowing recombinant MG53 protein to repair membrane damage when provided in the extracellular space. Recombinant human MG53 (rhMG53) protein purified from Escherichia coli fermentation provided dose-dependent protection against chemical, mechanical, or ultraviolet-induced damage to both muscle and nonmuscle cells. Injection of rhMG53 through multiple routes decreased muscle pathology in the mdx dystrophic mouse model. Our data support the concept of targeted cell membrane repair in regenerative medicine, and present MG53 protein as an attractive biological reagent for restoration of membrane repair defects in human diseases.
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Supplementary Material
Summary
Fig. S1. Recombinant human MG53 (rhMG53) can be isolated by several different approaches.
Fig. S2. Maintenance of body weight in mdx mice injected with rhMG53.
Fig. S3. Diaphragm muscle from mdx mice shows improved histopathology after subcutaneous injection of rhMG53.
Fig. S4. Pharmacokinetic properties of MBP-MG53 in wild-type mice.
Movie S1. MG53 localizes to sites of membrane damage after mechanical disruption.
Movie S2. BSA cannot prevent dye entry after UV laser–induced membrane damage to mouse skeletal muscle.
Movie S3. rhMG53 can prevent dye entry after UV laser–induced membrane damage to skeletal muscle.
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References and Notes
1
McNeil P. L., Steinhardt R. A., Plasma membrane disruption: Repair, prevention, adaptation. Annu. Rev. Cell Dev. Biol. 19, 697–731 (2003).
2
McNeil P. L., Kirchhausen T., An emergency response team for membrane repair. Nat. Rev. Mol. Cell Biol. 6, 499–505 (2005).
3
Steinhardt R. A., Bi G., Alderton J. M., Cell membrane resealing by a vesicular mechanism similar to neurotransmitter release. Science 263, 390–393 (1994).
4
McNeil P. L., Ito S., Gastrointestinal cell plasma membrane wounding and resealing in vivo. Gastroenterology 96, 1238–1248 (1989).
5
McNeil P. L., Khakee R., Disruptions of muscle fiber plasma membranes. Role in exercise-induced damage. Am. J. Pathol. 140, 1097–1109 (1992).
6
Clarke M. S., Caldwell R. W., Chiao H., Miyake K., McNeil P. L., Contraction-induced cell wounding and release of fibroblast growth factor in heart. Circ. Res. 76, 927–934 (1995).
7
Miyake K., McNeil P. L., Vesicle accumulation and exocytosis at sites of plasma membrane disruption. J. Cell Biol. 131, 1737–1745 (1995).
8
Togo T., Krasieva T. B., Steinhardt R. A., A decrease in membrane tension precedes successful cell-membrane repair. Mol. Biol. Cell 11, 4339–4346 (2000).
9
Bansal D., Miyake K., Vogel S. S., Groh S., Chen C. C., Williamson R., McNeil P. L., Campbell K. P., Defective membrane repair in dysferlin-deficient muscular dystrophy. Nature 423, 168–172 (2003).
10
van der Kooi A. J., Frankhuizen W. S., Barth P. G., Howeler C. J., Padberg G. W., Spaans F., Wintzen A. R., Wokke J. H., van Ommen G. J., de Visser M., Bakker E., Ginjaar H. B., Limb-girdle muscular dystrophy in the Netherlands: Gene defect identified in half the families. Neurology 68, 2125–2128 (2007).
11
Jaiswal J. K., Marlow G., Summerill G., Mahjneh I., Mueller S., Hill M., Miyake K., Haase H., Anderson L. V., Richard I., Kiuru-Enari S., McNeil P. L., Simon S. M., Bashir R., Patients with a non-dysferlin Miyoshi myopathy have a novel membrane repair defect. Traffic 8, 77–88 (2007).
12
Cooper S. T., Kizana E., Yates J. D., Lo H. P., Yang N., Wu Z. H., Alexander I. E., North K. N., Dystrophinopathy carrier determination and detection of protein deficiencies in muscular dystrophy using lentiviral MyoD-forced myogenesis. Neuromuscul. Disord. 17, 276–284 (2007).
13
Wenzel K., Geier C., Qadri F., Hubner N., Schulz H., Erdmann B., Gross V., Bauer D., Dechend R., Dietz R., Osterziel K. J., Spuler S., Ozcelik C., Dysfunction of dysferlin-deficient hearts. J. Mol. Med. 85, 1203–1214 (2007).
14
Chase T. H., Cox G. A., Burzenski L., Foreman O., Shultz L. D., Dysferlin deficiency and the development of cardiomyopathy in a mouse model of limb-girdle muscular dystrophy 2B. Am. J. Pathol. 175, 2299–2308 (2009).
15
Bazan N. G., Marcheselli V. L., Cole-Edwards K., Brain response to injury and neurodegeneration: Endogenous neuroprotective signaling. Ann. N. Y. Acad. Sci. 1053, 137–147 (2005).
16
Gajic O., Lee J., Doerr C. H., Berrios J. C., Myers J. L., Hubmayr R. D., Ventilator-induced cell wounding and repair in the intact lung. Am. J. Respir. Crit. Care Med. 167, 1057–1063 (2003).
17
Doherty K. R., McNally E. M., Repairing the tears: Dysferlin in muscle membrane repair. Trends Mol. Med. 9, 327–330 (2003).
18
Wang X., Xie W., Zhang Y., Lin P., Han L., Han P., Wang Y., Chen Z., Ji G., Zheng M., Weisleder N., Xiao R. P., Takeshima H., Ma J., Cheng H., Cardioprotection of ischemia/reperfusion injury by cholesterol-dependent MG53-mediated membrane repair. Circ. Res. 107, 76–83 (2010).
19
Cai C., Masumiya H., Weisleder N., Matsuda N., Nishi M., Hwang M., Ko J. K., Lin P., Thornton A., Zhao X., Pan Z., Komazaki S., Brotto M., Takeshima H., Ma J., MG53 nucleates assembly of cell membrane repair machinery. Nat. Cell Biol. 11, 56–64 (2009).
20
Cai C., Weisleder N., Ko J. K., Komazaki S., Sunada Y., Nishi M., Takeshima H., Ma J., Membrane repair defects in muscular dystrophy are linked to altered interaction between MG53, caveolin-3, and dysferlin. J. Biol. Chem. 284, 15894–15902 (2009).
21
Cao C. M., Zhang Y., Weisleder N., Ferrante C., Wang X., Lv F., Zhang Y., Song R., Hwang M., Jin L., Guo J., Peng W., Li G., Nishi M., Takeshima H., Ma J., Xiao R. P., MG53 constitutes a primary determinant of cardiac ischemic preconditioning. Circulation 121, 2565–2574 (2010).
22
Moser H., Duchenne muscular dystrophy: Pathogenetic aspects and genetic prevention. Hum. Genet. 66, 17–40 (1984).
23
Ray P. N., Belfall B., Duff C., Logan C., Kean V., Thompson M. W., Sylvester J. E., Gorski J. L., Schmickel R. D., Worton R. G., Cloning of the breakpoint of an X;21 translocation associated with Duchenne muscular dystrophy. Nature 318, 672–675 (1985).
24
Campbell K. P., Three muscular dystrophies: Loss of cytoskeleton-extracellular matrix linkage. Cell 80, 675–679 (1995).
25
Partridge T. A., Impending therapies for Duchenne muscular dystrophy. Curr. Opin. Neurol. 24, 415–422 (2011).
26
Watchko J., O’Day T., Wang B., Zhou L., Tang Y., Li J., Xiao X., Adeno-associated virus vector-mediated minidystrophin gene therapy improves dystrophic muscle contractile function in mdx mice. Hum. Gene Ther. 13, 1451–1460 (2002).
27
Yanagihara I., Inui K., Dickson G., Turner G., Piper T., Kaneda Y., Okada S., Expression of full-length human dystrophin cDNA in mdx mouse muscle by HVJ-liposome injection. Gene Ther. 3, 549–553 (1996).
28
Clemens P. R., Kochanek S., Sunada Y., Chan S., Chen H. H., Campbell K. P., Caskey C. T., In vivo muscle gene transfer of full-length dystrophin with an adenoviral vector that lacks all viral genes. Gene Ther. 3, 965–972 (1996).
29
Gussoni E., Pavlath G. K., Lanctot A. M., Sharma K. R., Miller R. G., Steinman L., Blau H. M., Normal dystrophin transcripts detected in Duchenne muscular dystrophy patients after myoblast transplantation. Nature 356, 435–438 (1992).
30
Gussoni E., Soneoka Y., Strickland C. D., Buzney E. A., Khan M. K., Flint A. F., Kunkel L. M., Mulligan R. C., Dystrophin expression in the mdx mouse restored by stem cell transplantation. Nature 401, 390–394 (1999).
31
Sampaolesi M., Blot S., D’Antona G., Granger N., Tonlorenzi R., Innocenzi A., Mognol P., Thibaud J. L., Galvez B. G., Barthélémy I., Perani L., Mantero S., Guttinger M., Pansarasa O., Rinaldi C., Cusella De Angelis M. G., Torrente Y., Bordignon C., Bottinelli R., Cossu G., Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. Nature 444, 574–579 (2006).
32
Nonaka I., Animal models of muscular dystrophies. Lab. Anim. Sci. 48, 8–17 (1998).
33
Zhu H., Lin P., De G., Choi K. H., Takeshima H., Weisleder N., Ma J., Polymerase transcriptase release factor (PTRF) anchors MG53 protein to cell injury site for initiation of membrane repair. J. Biol. Chem. 286, 12820–12824 (2011).
34
Lin P., Zhu H., Cai C., Wang X., Cao C., Xiao R., Pan Z., Weisleder N., Takeshima H., Ma J., Nonmuscle myosin IIA facilitates vesicle trafficking for MG53-mediated cell membrane repair. FASEB J. 26, 1875–1883 (2012).
35
Ng R., Metzger J. M., Claflin D. R., Faulkner J. A., Poloxamer 188 reduces the contraction-induced force decline in lumbrical muscles from mdx mice. Am. J. Physiol. Cell Physiol. 295, C146–C150 (2008).
36
Greenebaum B., Blossfield K., Hannig J., Carrillo C. S., Beckett M. A., Weichselbaum R. R., Lee R. C., Poloxamer 188 prevents acute necrosis of adult skeletal muscle cells following high-dose irradiation. Burns 30, 539–547 (2004).
37
Lee R. C., River L. P., Pan F. S., Ji L., Wollmann R. L., Surfactant-induced sealing of electropermeabilized skeletal muscle membranes in vivo. Proc. Natl. Acad. Sci. U.S.A. 89, 4524–4528 (1992).
38
Townsend D., Turner I., Yasuda S., Martindale J., Davis J., Shillingford M., Kornegay J. N., Metzger J. M., Chronic administration of membrane sealant prevents severe cardiac injury and ventricular dilatation in dystrophic dogs. J. Clin. Invest. 120, 1140–1150 (2010).
39
Yasuda S., Townsend D., Michele D. E., Favre E. G., Day S. M., Metzger J. M., Dystrophic heart failure blocked by membrane sealant poloxamer. Nature 436, 1025–1029 (2005).
40
Follis F., Jenson B., Blisard K., Hall E., Wong R., Kessler R., Temes T., Wernly J., Role of poloxamer 188 during recovery from ischemic spinal cord injury: A preliminary study. J. Invest. Surg. 9, 149–156 (1996).
41
Frim D. M., Wright D. A., Curry D. J., Cromie W., Lee R., Kang U. J., The surfactant poloxamer-188 protects against glutamate toxicity in the rat brain. Neuroreport 15, 171–174 (2004).
42
Kilinc D., Gallo G., Barbee K., Poloxamer 188 reduces axonal beading following mechanical trauma to cultured neurons. Conf. Proc. IEEE Eng. Med. Biol. Soc. 2007, 5395–5398 (2007).
43
Gulik-Krzywicki T., Balerna M., Vincent J. P., Lazdunski M., Freeze-fracture study of cardiotoxin action on axonal membrane and axonal membrane lipid vesicles. Biochim. Biophys. Acta 643, 101–114 (1981).
44
Rivas E. A., Le Maire M., Gulik-Krzywicki T., Isolation of rhodopsin by the combined action of cardiotoxin and phospholipase A2 on rod outer segment membranes. Biochim. Biophys. Acta 644, 127–133 (1981).
45
Dressler V., Schwister K., Haest C. W., Deuticke B., Dielectric breakdown of the erythrocyte membrane enhances transbilayer mobility of phospholipids. Biochim. Biophys. Acta 732, 304–307 (1983).
46
Fujiwara T., Ritchie K., Murakoshi H., Jacobson K., Kusumi A., Phospholipids undergo hop diffusion in compartmentalized cell membrane. J. Cell Biol. 157, 1071–1081 (2002).
47
Sonnleitner A., Schütz G. J., Schmidt T., Free Brownian motion of individual lipid molecules in biomembranes. Biophys. J. 77, 2638–2642 (1999).
48
Bansal D., Campbell K. P., Dysferlin and the plasma membrane repair in muscular dystrophy. Trends Cell Biol. 14, 206–213 (2004).
49
Fong P. Y., Turner P. R., Denetclaw W. F., Steinhardt R. A., Increased activity of calcium leak channels in myotubes of Duchenne human and mdx mouse origin. Science 250, 673–676 (1990).
50
Deconinck N., Dan B., Pathophysiology of duchenne muscular dystrophy: Current hypotheses. Pediatr. Neurol. 36, 1–7 (2007).
51
Pasternak C., Wong S., Elson E. L., Mechanical function of dystrophin in muscle cells. J. Cell Biol. 128, 355–361 (1995).
52
Yokota T., Lu Q. L., Partridge T., Kobayashi M., Nakamura A., Takeda S., Hoffman E., Efficacy of systemic morpholino exon-skipping in Duchenne dystrophy dogs. Ann. Neurol. 65, 667–676 (2009).
53
Welch E. M., Barton E. R., Zhuo J., Tomizawa Y., Friesen W. J., Trifillis P., Paushkin S., Patel M., Trotta C. R., Hwang S., Wilde R. G., Karp G., Takasugi J., Chen G., Jones S., Ren H., Moon Y. C., Corson D., Turpoff A. A., Campbell J. A., Conn M. M., Khan A., Almstead N. G., Hedrick J., Mollin A., Risher N., Weetall M., Yeh S., Branstrom A. A., Colacino J. M., Babiak J., Ju W. D., Hirawat S., Northcutt V. J., Miller L. L., Spatrick P., He F., Kawana M., Feng H., Jacobson A., Peltz S. W., Sweeney H. L., PTC124 targets genetic disorders caused by nonsense mutations. Nature 447, 87–91 (2007).
54
Allen D. G., Whitehead N. P., Duchenne muscular dystrophy—What causes the increased membrane permeability in skeletal muscle? Int. J. Biochem. Cell Biol. 43, 290–294 (2011).
55
Rooney J. E., Gurpur P. B., Burkin D. J., Laminin-111 protein therapy prevents muscle disease in the mdx mouse model for Duchenne muscular dystrophy. Proc. Natl. Acad. Sci. U.S.A. 106, 7991–7996 (2009).
56
Sonnemann K. J., Heun-Johnson H., Turner A. J., Baltgalvis K. A., Lowe D. A., Ervasti J. M., Functional substitution by TAT-utrophin in dystrophin-deficient mice. PLoS Med. 6, e1000083 (2009).
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Published In
Science Translational Medicine
Volume 4 | Issue 139
June 2012
June 2012
Copyright
Copyright © 2012, American Association for the Advancement of Science.
Submission history
Received: 22 February 2012
Accepted: 11 May 2012
Acknowledgments
We thank Zelgen-Biopharm for their assistance in developing methodology for the scale-up production of recombinant MG53 proteins. Funding: This work was supported by grants from the NIH to N.W. (AR063084) and J.M. (AG015556, HL069000, and AR061385), a developmental grant from the Jain Foundation and an NIH Small Business Innovation Research grant (AR060019) to TRIM-edicine, and the Tissue Analytic Services Shared Resource of The Cancer Institute of New Jersey (P30CA072720). Author contributions: N.W., X.W., and J.M. developed the concept for the studies. N.W. and J.M. designed the experiments. N.T., P.L., M.T., T.T., C.F., R.Y., P.-J.C., X.Z., and M.H. participated in production of recombinant MG53 proteins. N.W., N.T., C.F., and P.-J.C. developed methods to measure rhMG53 in cell-based assays. N.T., X.W., C. Cao, Y.Z., P.L., H.Z., P.-J.C., M.S., C. Cai, H.C., H.T., and R.-P.X. performed in vitro assays of MG53 function. N.W., N.T., T.T., C.F., H.Z., R.Y., X.Z., and H.T. performed in vivo assays. N.T., T.T., and R.Y. performed the toxicological assessments. N.W. and J.M. wrote the manuscript. Competing interests: N.W. and J.M. have an equity interest in TRIM-edicine, which develops rhMG53 for treatment of human diseases. Patents on the use of rhMG53 are held by the University of Medicine and Dentistry of New Jersey. Data and materials availability: Access to rhMG53 protein can be obtained from TRIM-edicine Inc. after establishment of a Material Transfer Agreement.
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