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Muscle-derived cell-mediated ex vivo gene therapy for urological dysfunction

Gene Therapy volume 9, pages 1617–1626 (2002)Cite this article

Abstract

We have tested the feasibility of muscle-based gene therapy and tissue engineering for urological dysfunction using highly purified muscle-derived cells (MDC) that display stem cell characteristics. We then explored the potential use of these MDC as an alternative therapy for the treatment of impaired detrusor contractility. The MDC were genetically engineered to express the gene encoding β-galactosidase and injected into the bladder walls of SCID mice. The injected bladders were harvested at various time-points after injection and assayed for β-galactosidase activity; the presence of myofibers within the injected tissue was determined by detection of fast myosin heavy chain isoform (MyHCs). We have demonstrated that the injected MDC are capable of not only surviving in the lower urinary tract, but also improving the contractility of the bladder following an induced injury. Two potential mechanisms can be used to explain this finding. First, we have observed that some of the β-galactosidase-expressing cells expressed α-smooth muscle actin, suggesting a differentiation into smooth muscle. Second, a stain for acetylcholine receptors (AChRs), which identifies the location of neuromuscular junctions, revealed that the myofibers derived from the doner cells became innervated into the bladder as early as 2 weeks after injection. These results suggest that gene therapy and tissue engineering based on MDC potentially can be used for urological dysfunction.

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References

  1. Agency for Health Care Policy and Research. Public Health Service: Urinary Incontinence, Public Information, US Department of Health and Human Services. December 1996.

  2. Gelber DA, Good DC, Laven LJ, Verhulst SJ . Causes of urinary incontinence after acute hemispheric stroke Stroke 1993 24: 378–382

    Article  CAS  Google Scholar 

  3. Gelber DA, Good DC, Laven LJ, Verhulst SJ . Causes of urinary incontinence after acute hemispheric stroke Chancellor MB, Blaivas JG. Atlas of Urodynamics. Williams and Wilkins: Philadelphia, 1996.

  4. Nakayama H et al. Prevalence and risk factors of incontinence after stroke. The Copenhagen Stroke Study Stroke 1997 28: 58–62

    Article  CAS  Google Scholar 

  5. Wein AJ, Raezer D, Malloy T . Failure of the bethanechol supersensitivity test to predict improved voiding after subcutaneous bethanechol administration J Urol 1980 123: 302

    Google Scholar 

  6. Wein AJ . Pharmacology of incontinence Urol Clin North Am 1995 22: 557–577

    CAS  PubMed  Google Scholar 

  7. Gussoni E et al. Dystrophin expression in the mdx mouse restored by stem cell transplantation Nature 1999 401: 390–394

    CAS  Google Scholar 

  8. Musgrave DM, Huard J . Muscle-based tissue engineering for the musculoskeletal system Gene Ther Mol Bio 1999 3: 207–221

    Google Scholar 

  9. Lee JY et al. Clonal isolation of muscle-derived cells capable of enhancing muscle regeneration and bone healing J Cell Biol 2000 150: 1085–1099

    Article  CAS  Google Scholar 

  10. Martinek V, Fu FH, Huard J . Gene therapy and tissue engineering in sports medicine Phys Sports Med 2000 28: 34–51

    Article  CAS  Google Scholar 

  11. Partridge TA et al. Conversion of mdx myofibers from dystrophin negative to positive by injection of normal myoblasts Nature 1989 337: 176–179

    Article  CAS  Google Scholar 

  12. Karpati G et al. Myoblast transfer in Duchenne muscular dystrophy Ann Neurol 1993 34: 8–17

    Article  CAS  Google Scholar 

  13. Tremblay JP et al. Results of a blind clinical study of myoblast transplantation without immunosuppressive treatment in young boys with DMD Cell Transplantation 1993 2: 99–112

    Article  CAS  Google Scholar 

  14. Beauchamp JR, Morgan JE, Pagel CN, Partridge TA . Quantitative studies of efficacy of myoblast transplantation Muscle Nerve 1994 18: S261

    Google Scholar 

  15. Huard J et al. Gene transfer into skeletal muscles by isogenic myoblasts Hum Gene Ther 1994 5: 949–958

    Article  CAS  Google Scholar 

  16. Huard J et al. High efficiency of muscle regeneration following human myoblast clone transplantation in SCID mice J Clin Invest 1994 93: 586–599

    Article  CAS  Google Scholar 

  17. Mendell JR et al. Myoblast transfer in the treatment of Duchenne's muscular dystrophy New Engl J Med 1995 333: 832–838

    Article  CAS  Google Scholar 

  18. Fan Y, Maley M, Beilharz M, Grounds M . Rapid death of injected myoblasts in myoblast transfer therapy Muscle Nerve 1996 19: 853–860

    Article  CAS  Google Scholar 

  19. Gussoni E, Blau HM, Kunkel LM . The fate of individual myoblasts after transplantation into muscles of DMD patients Nat Med 1997 3: 970–977

    Article  CAS  Google Scholar 

  20. Qu Z et al. Development of approaches to improve cell survival in myoblast transfer therapy J Cell Biol 1998 142: 1257–1267

    Article  CAS  Google Scholar 

  21. Richler C, Yaffe D . The in vitro cultivation and differentiation capacities of myogenic cell lines Dev Biol 1970 23: 1–22

    Article  CAS  Google Scholar 

  22. Rando TA, Blau HM . Primary mouse myoblast purification, characterization, and transplantation for cell-mediated gene therapy J Cell Biol 1994 125: 1275–1287

    Article  CAS  Google Scholar 

  23. Beauchamp JR, Morgan JE, Pagel CN, Partridge TA . Dynamics of myoblast transplantation reveal a discrete minority of precursors with stem cell-like properties as the myogenic source J Cell Biol 1999 144: 1113–1122

    Article  CAS  Google Scholar 

  24. Katagiri T et al. Bone morphogenetic protein-2 converts the differentiation pathway of C2C12 myoblasts into the osteoblast lineage J Cell Biol 1995 127: 1755–1766

    Article  Google Scholar 

  25. Baroffio A et al. Identification of self-renewing myoblasts in the progeny of single human muscle satellite cells Differentiation 1996 60: 47–57

    Article  CAS  Google Scholar 

  26. Cornelison DD, Wold BJ . Single-cell analysis of regulatory gene expression in quiescent and activated mouse skeletal muscle satellite cells Dev Biol 1997 191: 270–283

    Article  CAS  Google Scholar 

  27. Miller JB, Schaefer L, Dominov JA . Seeking muscle stem cells Curr Top Dev Biol 1999 43: 191–219

    Article  CAS  Google Scholar 

  28. Jackson KA, Mi T, Goodell MA . Hematopoietic potential of stem cells isolated from murine skeletal muscle Proc Natl Acad Sci USA 1999 96: 14482–14486

    Article  CAS  Google Scholar 

  29. Bosch P et al. Osteoprogenitor cells within skeletal muscle J Orthop Res 2000 18: 933–944

    Article  CAS  Google Scholar 

  30. McHugh KM . Molecular analysis of smooth muscle development in the mouse Dev Dyn 1995 204: 278–290

    Article  CAS  Google Scholar 

  31. Darby I, Skalli O, Gabbiani G . Alpha-smooth muscle actin is transiently expressed by myofibroblasts during experimental wound healing Lab Invest 1990 63: 21–29

    CAS  Google Scholar 

  32. Shimizu T, Matsumura K, Sunada Y, Mannen T . Dense immunostaining on both neuromuscular and myotendon junctions with an antidystrophin antibody Biomed Res 1989 10: 405–409

    Article  CAS  Google Scholar 

  33. Shortliffe LM et al. Treatment of urinary incontinence by the periurethral implantation of glutaraldehyde cross-linked collagen J Urol 1989 141: 538–541

    Article  CAS  Google Scholar 

  34. McGuire EJ, Appell R . Transurethral collagen injection for urinary incontinence Urology 1994 43: 413–415

    Article  CAS  Google Scholar 

  35. Frey P, Berger D, Jenny P, Herzog B . Suburethral collagen injection for the endoscopic treatment of vesicoureteral reflux in children: follow-up study of 97 treated ureters and histological analysis of collagen implants J Urol 1992 148: 718–723

    Article  CAS  Google Scholar 

  36. Herschorn S, Steele DJ, Radomski SB . Follow up of intraurethral collagen for female stress urinary incontinence J Urol 1996 156: 1305–1309

    Article  CAS  Google Scholar 

  37. Yokoyama T et al. Persistence and survival of autologous muscle-derived cells versus bovine collagen as a potential treatment of stress urinary incontinence J Urol 2001 165: 271–276

    Article  CAS  Google Scholar 

  38. Shimizu RT et al. The smooth muscle α-actin gene promoter is differentially regulated in smooth muscle versus non-smooth muscle cells J Biol Chem 1995 270: 7631–7643

    Article  CAS  Google Scholar 

  39. Gibson AJ . Dermal fibroblasts convert to a myogenic lineage in mdx mouse muscle J Cell Sci 1995 108: 207–214

    CAS  PubMed  Google Scholar 

  40. Gibson AJ . Dermal fibroblasts convert to a myogenic lineage in mdx mouse muscle Li Y, Huard J. Differentiation of muscle-derived cells into myofibroblasts in injured skeletal muscle. Am J Pathol 2002 (in press).

  41. Qu-Petersen Z et al. Identification of a novel population of muscle stem cells in mice: potential for muscle regeneration J Cell Biol 2002 157: 851–864

    Article  CAS  Google Scholar 

  42. Floyd SS et al. Ex vivo gene transfer using adenovirus-mediated full-length dystrophin delivery to dystrophic muscles Gene Therapy 1998 5: 19–30

    Article  CAS  Google Scholar 

  43. Yuasa K et al. Effective restoration of dystrophin associated proteins in vivo by adenovirus-mediated transfer of truncated dystrophin cDNAs FEBS Lett 1998 425: 329–336

    Article  CAS  Google Scholar 

  44. Yuasa K et al. Effective restoration of dystrophin associated proteins in vivo by adenovirus-mediated transfer of truncated dystrophin cDNAs Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning. A Laboratory Manual. Cold Spring Harbar Laboratory Press: Plainview, NY, 1989, pp 16.66-16.67.

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Acknowledgements

The authors wish to acknowledge Marcelle Pellerin, Rudrani Ghosh and Maggie Sistek for their technical and secretarial assistance, respectively, and Ryan Sauder for editorial assistance. The authors wish also to acknowledge the critical comments made by Mr Ron J Jankowski and Dr Shing-Hwa Lu. This work was supported by William F and Jean W Donaldson Chair, NIDDK support: NIH RO1 DK55387 and NIH 2R01 DK05538705, and Pittsburgh Tissue Engineering Initiative (PTEI).

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Authors and Affiliations

  1. Department of Urology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    T Yokoyama, J Y Lee & M B Chancellor

  2. Department of Pharmacology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    G T Somogyi & W C de Groat

  3. Department of Orthopaedic Surgery, and Molecular Genetics and Biochemistry, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    J Huard, R Pruchnic, Z Qu & Y Li

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  1. J Huard
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Huard, J., Yokoyama, T., Pruchnic, R. et al. Muscle-derived cell-mediated ex vivo gene therapy for urological dysfunction. Gene Ther 9, 1617–1626 (2002). https://doi.org/10.1038/sj.gt.3301816

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