Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1995 Aug;96(2):1137–1144. doi: 10.1172/JCI118101

Expression of transforming growth factor-beta 1 in dystrophic patient muscles correlates with fibrosis. Pathogenetic role of a fibrogenic cytokine.

P Bernasconi 1, E Torchiana 1, P Confalonieri 1, R Brugnoni 1, R Barresi 1, M Mora 1, F Cornelio 1, L Morandi 1, R Mantegazza 1
PMCID: PMC185304  PMID: 7635950

Abstract

Duchenne muscular dystrophy is a fatal disorder characterized by progressive muscular weakness, wasting, and severe muscle contractures in later disease stages. Muscle biopsy reveals conspicuous myofiber degeneration and fibrosis substituting muscle tissue. We quantitatively determined mRNA of the potent fibrogenic cytokine transforming growth factor-beta 1 by quantitative PCR in 15 Duchenne muscular dystrophy, 13 Becker muscular dystrophy, 11 spinal muscular atrophy patients, and 16 controls. Higher transforming growth factor-beta 1 expression was greater in Duchenne muscular dystrophy patients than controls (P = 0.012) and Becker patients (P = 0.03). Fibrosis was significantly more prominent in Duchenne muscular dystrophy than Becker muscular dystrophy, spinal muscular atrophy, and controls. The proportion of connective tissue in muscle biopsies increased progressively with age in Duchenne muscular dystrophy patients, while transforming growth factor-beta 1 levels peaked at 2 and 6 yr of age. Transforming growth factor-beta 1 protein was also detected by immunocytochemistry and immunoblotting. Our findings suggest that transforming growth factor-beta 1 stimulates fibrosis in Duchenne muscular dystrophy. Expression of transforming growth factor-beta 1 in the early stages of Duchenne muscular dystrophy may be critical in initiating muscle fibrosis and antifibrosis treatment could slow progression of the disease, increasing the utility of gene therapy.

Full text

PDF

Images in this article

1138Image
on p.1138
1138Image
on p.1138
1139Image
on p.1139
1141Image
on p.1141
1141Image
on p.1141
1141Image
on p.1141
1141Image
on p.1141
1141Image
on p.1141
1141Image
on p.1141
1141Image
on p.1141
1142Image
on p.1142

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arahata K., Engel A. G. Monoclonal antibody analysis of mononuclear cells in myopathies. I: Quantitation of subsets according to diagnosis and sites of accumulation and demonstration and counts of muscle fibers invaded by T cells. Ann Neurol. 1984 Aug;16(2):193–208. doi: 10.1002/ana.410160206. [DOI] [PubMed] [Google Scholar]
  2. Border W. A., Noble N. A. Transforming growth factor beta in tissue fibrosis. N Engl J Med. 1994 Nov 10;331(19):1286–1292. doi: 10.1056/NEJM199411103311907. [DOI] [PubMed] [Google Scholar]
  3. Border W. A., Okuda S., Languino L. R., Sporn M. B., Ruoslahti E. Suppression of experimental glomerulonephritis by antiserum against transforming growth factor beta 1. Nature. 1990 Jul 26;346(6282):371–374. doi: 10.1038/346371a0. [DOI] [PubMed] [Google Scholar]
  4. Brandes M. E., Allen J. B., Ogawa Y., Wahl S. M. Transforming growth factor beta 1 suppresses acute and chronic arthritis in experimental animals. J Clin Invest. 1991 Mar;87(3):1108–1113. doi: 10.1172/JCI115073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Broekelmann T. J., Limper A. H., Colby T. V., McDonald J. A. Transforming growth factor beta 1 is present at sites of extracellular matrix gene expression in human pulmonary fibrosis. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6642–6646. doi: 10.1073/pnas.88.15.6642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Castilla A., Prieto J., Fausto N. Transforming growth factors beta 1 and alpha in chronic liver disease. Effects of interferon alfa therapy. N Engl J Med. 1991 Apr 4;324(14):933–940. doi: 10.1056/NEJM199104043241401. [DOI] [PubMed] [Google Scholar]
  7. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  8. Cornelio F., Dones I. Muscle fiber degeneration and necrosis in muscular dystrophy and other muscle diseases: cytochemical and immunocytochemical data. Ann Neurol. 1984 Dec;16(6):694–701. doi: 10.1002/ana.410160612. [DOI] [PubMed] [Google Scholar]
  9. D'Amore P. A., Brown R. H., Jr, Ku P. T., Hoffman E. P., Watanabe H., Arahata K., Ishihara T., Folkman J. Elevated basic fibroblast growth factor in the serum of patients with Duchenne muscular dystrophy. Ann Neurol. 1994 Mar;35(3):362–365. doi: 10.1002/ana.410350320. [DOI] [PubMed] [Google Scholar]
  10. Dasch J. R., Pace D. R., Waegell W., Inenaga D., Ellingsworth L. Monoclonal antibodies recognizing transforming growth factor-beta. Bioactivity neutralization and transforming growth factor beta 2 affinity purification. J Immunol. 1989 Mar 1;142(5):1536–1541. [PubMed] [Google Scholar]
  11. Duance V. C., Stephens H. R., Dunn M., Bailey A. J., Dubowitz V. A role for collagen in the pathogenesis of muscular dystrophy? Nature. 1980 Apr 3;284(5755):470–472. doi: 10.1038/284470a0. [DOI] [PubMed] [Google Scholar]
  12. Friedman S. L. Seminars in medicine of the Beth Israel Hospital, Boston. The cellular basis of hepatic fibrosis. Mechanisms and treatment strategies. N Engl J Med. 1993 Jun 24;328(25):1828–1835. doi: 10.1056/NEJM199306243282508. [DOI] [PubMed] [Google Scholar]
  13. Galli S. J. New concepts about the mast cell. N Engl J Med. 1993 Jan 28;328(4):257–265. doi: 10.1056/NEJM199301283280408. [DOI] [PubMed] [Google Scholar]
  14. Giri S. N., Hyde D. M., Hollinger M. A. Effect of antibody to transforming growth factor beta on bleomycin induced accumulation of lung collagen in mice. Thorax. 1993 Oct;48(10):959–966. doi: 10.1136/thx.48.10.959. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Gorospe J. R., Tharp M. D., Hinckley J., Kornegay J. N., Hoffman E. P. A role for mast cells in the progression of Duchenne muscular dystrophy? Correlations in dystrophin-deficient humans, dogs, and mice. J Neurol Sci. 1994 Mar;122(1):44–56. doi: 10.1016/0022-510x(94)90050-7. [DOI] [PubMed] [Google Scholar]
  16. Gorospe J. R., Tharp M., Demitsu T., Hoffman E. P. Dystrophin-deficient myofibers are vulnerable to mast cell granule-induced necrosis. Neuromuscul Disord. 1994 Jul;4(4):325–333. doi: 10.1016/0960-8966(94)90068-x. [DOI] [PubMed] [Google Scholar]
  17. Hoffman E. P., Brown R. H., Jr, Kunkel L. M. Dystrophin: the protein product of the Duchenne muscular dystrophy locus. Cell. 1987 Dec 24;51(6):919–928. doi: 10.1016/0092-8674(87)90579-4. [DOI] [PubMed] [Google Scholar]
  18. Jullien P., Berg T. M., Lawrence D. A. Acidic cellular environments: activation of latent TGF-beta and sensitization of cellular responses to TGF-beta and EGF. Int J Cancer. 1989 May 15;43(5):886–891. doi: 10.1002/ijc.2910430525. [DOI] [PubMed] [Google Scholar]
  19. Kim S. J., Park K., Koeller D., Kim K. Y., Wakefield L. M., Sporn M. B., Roberts A. B. Post-transcriptional regulation of the human transforming growth factor-beta 1 gene. J Biol Chem. 1992 Jul 5;267(19):13702–13707. [PubMed] [Google Scholar]
  20. Koenig M., Beggs A. H., Moyer M., Scherpf S., Heindrich K., Bettecken T., Meng G., Müller C. R., Lindlöf M., Kaariainen H. The molecular basis for Duchenne versus Becker muscular dystrophy: correlation of severity with type of deletion. Am J Hum Genet. 1989 Oct;45(4):498–506. [PMC free article] [PubMed] [Google Scholar]
  21. Kovacs E. J. Fibrogenic cytokines: the role of immune mediators in the development of scar tissue. Immunol Today. 1991 Jan;12(1):17–23. doi: 10.1016/0167-5699(91)90107-5. [DOI] [PubMed] [Google Scholar]
  22. Li B., Sehajpal P. K., Khanna A., Vlassara H., Cerami A., Stenzel K. H., Suthanthiran M. Differential regulation of transforming growth factor beta and interleukin 2 genes in human T cells: demonstration by usage of novel competitor DNA constructs in the quantitative polymerase chain reaction. J Exp Med. 1991 Nov 1;174(5):1259–1262. doi: 10.1084/jem.174.5.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Logan A., Berry M., Gonzalez A. M., Frautschy S. A., Sporn M. B., Baird A. Effects of transforming growth factor beta 1 on scar production in the injured central nervous system of the rat. Eur J Neurosci. 1994 Mar 1;6(3):355–363. doi: 10.1111/j.1460-9568.1994.tb00278.x. [DOI] [PubMed] [Google Scholar]
  24. Lyons R. M., Gentry L. E., Purchio A. F., Moses H. L. Mechanism of activation of latent recombinant transforming growth factor beta 1 by plasmin. J Cell Biol. 1990 Apr;110(4):1361–1367. doi: 10.1083/jcb.110.4.1361. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Mantegazza R., Andreetta F., Bernasconi P., Baggi F., Oksenberg J. R., Simoncini O., Mora M., Cornelio F., Steinman L. Analysis of T cell receptor repertoire of muscle-infiltrating T lymphocytes in polymyositis. Restricted V alpha/beta rearrangements may indicate antigen-driven selection. J Clin Invest. 1993 Jun;91(6):2880–2886. doi: 10.1172/JCI116533. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Massagué J., Attisano L., Wrana J. L. The TGF-beta family and its composite receptors. Trends Cell Biol. 1994 May;4(5):172–178. doi: 10.1016/0962-8924(94)90202-x. [DOI] [PubMed] [Google Scholar]
  27. Massagué J., Cheifetz S., Laiho M., Ralph D. A., Weis F. M., Zentella A. Transforming growth factor-beta. Cancer Surv. 1992;12:81–103. [PubMed] [Google Scholar]
  28. McDouall R. M., Dunn M. J., Dubowitz V. Nature of the mononuclear infiltrate and the mechanism of muscle damage in juvenile dermatomyositis and Duchenne muscular dystrophy. J Neurol Sci. 1990 Nov;99(2-3):199–217. doi: 10.1016/0022-510x(90)90156-h. [DOI] [PubMed] [Google Scholar]
  29. Mendell J. R., Moxley R. T., Griggs R. C., Brooke M. H., Fenichel G. M., Miller J. P., King W., Signore L., Pandya S., Florence J. Randomized, double-blind six-month trial of prednisone in Duchenne's muscular dystrophy. N Engl J Med. 1989 Jun 15;320(24):1592–1597. doi: 10.1056/NEJM198906153202405. [DOI] [PubMed] [Google Scholar]
  30. Morandi L., Mora M., Bernasconi P., Mantegazza R., Gebbia M., Balestrini M. R., Cornelio F. Very small dystrophin molecule in a family with a mild form of Becker dystrophy. Neuromuscul Disord. 1993 Jan;3(1):65–70. doi: 10.1016/0960-8966(93)90043-j. [DOI] [PubMed] [Google Scholar]
  31. Morandi L., Mora M., Gussoni E., Tedeschi S., Cornelio F. Dystrophin analysis in Duchenne and Becker muscular dystrophy carriers: correlation with intracellular calcium and albumin. Ann Neurol. 1990 Nov;28(5):674–679. doi: 10.1002/ana.410280512. [DOI] [PubMed] [Google Scholar]
  32. Roberts A. B., Sporn M. B., Assoian R. K., Smith J. M., Roche N. S., Wakefield L. M., Heine U. I., Liotta L. A., Falanga V., Kehrl J. H. Transforming growth factor type beta: rapid induction of fibrosis and angiogenesis in vivo and stimulation of collagen formation in vitro. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4167–4171. doi: 10.1073/pnas.83.12.4167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Shah M., Foreman D. M., Ferguson M. W. Control of scarring in adult wounds by neutralising antibody to transforming growth factor beta. Lancet. 1992 Jan 25;339(8787):213–214. doi: 10.1016/0140-6736(92)90009-r. [DOI] [PubMed] [Google Scholar]
  34. Sharma K. R., Mynhier M. A., Miller R. G. Cyclosporine increases muscular force generation in Duchenne muscular dystrophy. Neurology. 1993 Mar;43(3 Pt 1):527–532. doi: 10.1212/wnl.43.3_part_1.527. [DOI] [PubMed] [Google Scholar]
  35. Shull M. M., Ormsby I., Kier A. B., Pawlowski S., Diebold R. J., Yin M., Allen R., Sidman C., Proetzel G., Calvin D. Targeted disruption of the mouse transforming growth factor-beta 1 gene results in multifocal inflammatory disease. Nature. 1992 Oct 22;359(6397):693–699. doi: 10.1038/359693a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Siebert P. D., Larrick J. W. Competitive PCR. Nature. 1992 Oct 8;359(6395):557–558. doi: 10.1038/359557a0. [DOI] [PubMed] [Google Scholar]
  37. Sporn M. B., Roberts A. B. Transforming growth factor-beta: recent progress and new challenges. J Cell Biol. 1992 Dec;119(5):1017–1021. doi: 10.1083/jcb.119.5.1017. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Wahl S. M., Allen J. B., Costa G. L., Wong H. L., Dasch J. R. Reversal of acute and chronic synovial inflammation by anti-transforming growth factor beta. J Exp Med. 1993 Jan 1;177(1):225–230. doi: 10.1084/jem.177.1.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Wahl S. M., Allen J. B., Wong H. L., Dougherty S. F., Ellingsworth L. R. Antagonistic and agonistic effects of transforming growth factor-beta and IL-1 in rheumatoid synovium. J Immunol. 1990 Oct 15;145(8):2514–2519. [PubMed] [Google Scholar]
  40. Wahl S. M., McCartney-Francis N., Mergenhagen S. E. Inflammatory and immunomodulatory roles of TGF-beta. Immunol Today. 1989 Aug;10(8):258–261. doi: 10.1016/0167-5699(89)90136-9. [DOI] [PubMed] [Google Scholar]
  41. Wahl S. M. Transforming growth factor beta: the good, the bad, and the ugly. J Exp Med. 1994 Nov 1;180(5):1587–1590. doi: 10.1084/jem.180.5.1587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Wang A. M., Doyle M. V., Mark D. F. Quantitation of mRNA by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9717–9721. doi: 10.1073/pnas.86.24.9717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Wolf Y. G., Rasmussen L. M., Ruoslahti E. Antibodies against transforming growth factor-beta 1 suppress intimal hyperplasia in a rat model. J Clin Invest. 1994 Mar;93(3):1172–1178. doi: 10.1172/JCI117070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Yamazaki M., Minota S., Sakurai H., Miyazono K., Yamada A., Kanazawa I., Kawai M. Expression of transforming growth factor-beta 1 and its relation to endomysial fibrosis in progressive muscular dystrophy. Am J Pathol. 1994 Feb;144(2):221–226. [PMC free article] [PubMed] [Google Scholar]
  45. Zentella A., Massagué J. Transforming growth factor beta induces myoblast differentiation in the presence of mitogens. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):5176–5180. doi: 10.1073/pnas.89.11.5176. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES