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In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy

Gene Therapy volume 6, pages 1267–1275 (1999)Cite this article

Abstract

Gene-based therapies may provide a way to treat inherited skin disorders but current approaches suffer serious limitations. The surgical procedures required to transplant ex vivo modified keratinocytes are likely to result in scarring and contracture, thereby limiting the area that can be treated. In addition, none of the methods currently available for in vivo gene transfer to epidermis leads to long-term transgene expression. The goal of this study was to develop a means for in vivo gene transfer to epidermis that would result in long-term transgene expression. We report here the first successful in vivo gene transfer that results in sustained transgene expression in epidermis. Hyperplastic mouse skin was transduced by direct injection of VSV-G pseudotyped retroviral vectors encoding the LacZ reporter gene. In mice tolerant to β-galactosidase (β-gal), transgene expression was noted in hair follicles and interfollicular epidermis for the duration of the experiment (16 weeks after transduction). Based on the kinetics of epidermal turnover in mouse skin, expression for this length of time strongly suggests stem cell transduction. In immunocompetent mice intolerant to β-gal, transgene expression was lost by 3 weeks after transduction, concurrent with the onset of host immune responses to the transgene product.

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References

  1. Ghazizadeh S, Kolodka TM, Taichman LB . The skin as a vehicle for gene therapy. In: Jameson JL (ed) Principles of Molecular Medicine Humana Press: Totowa, NJ 1998 775–779

    Chapter  Google Scholar 

  2. Krueger GG et al. Genetically modified skin to treat disease: potential and limitations J Invest Dermatol 1995 103: 76S–84S

    Article  Google Scholar 

  3. Vogel JC, Walker PS, Hengge UR . Gene therapy for skin diseases Adv Dermatol 1996 11: 383–398

    CAS  PubMed  Google Scholar 

  4. Khavari PA, Krueger GG . Cutaneous gene therapy Adv Clin Res 1997 15: 27–35

    CAS  Google Scholar 

  5. Hengge UR et al. Cytokine gene expression in epidermis with biological effects following injection of naked DNA Nature 1995 10: 161–166

    CAS  Google Scholar 

  6. Li L, Hoffman RM . The feasibility of targeted selective gene therapy of the hair follicle Nature 1995 1: 705–706

    Google Scholar 

  7. Alexander MY, Akhurst RJ . Liposome-mediated gene transfer and expression via the skin Hum Molec Gen 1995 4: 2279–2285

    Article  CAS  Google Scholar 

  8. Setoguchi Y, Jaffe HA, Danel C, Crystal RG . Ex vivo and in vivo gene transfer to the skin using replication-deficient recombinant adenovirus vectors J Invest Dermatol 1994 102: 415–421

    Article  CAS  Google Scholar 

  9. Andree C et al. In vivo transfer and expression of a human epidermal growth factor gene accelerates wound repair Proc Natl Acad Sci USA 1994 91: 12188–12192

    Article  CAS  Google Scholar 

  10. Mackenzie IC . Retroviral transduction of murine epidermal stem cells demonstrates clonal units of epidermal structure J Invest Dermatol 1997 109: 377–383

    Article  CAS  Google Scholar 

  11. Potten CS . Cell replacement in epidermis (keratopoiesis) via discrete units of proliferation Int Rev Cytol 1981 69: 271–318

    Article  CAS  Google Scholar 

  12. Jones PH . Isolation and characterization of human epidermal stem cells Clin Sci 1996 91: 141–146

    Article  CAS  Google Scholar 

  13. Jolly D . Viral vector systems for gene therapy. In: Sobol RE, Scanlon KJ (eds) The Internet Book of Gene Therapy Appleton & Lange: Stamford, CT 1997 3–16

    Google Scholar 

  14. Miller DG, Adam MA, Miller AD . Gene transfer by retrovirus vectors occurs only in cells that are actively replicating at the time of infection Mol Cell Biol 1990 10: 4239–4242

    Article  CAS  Google Scholar 

  15. Al-Barwari SE, Potten CS . Regeneration and dose-response characteristics of irradiated mouse dorsal epidermal cells Int J Radiat Biol 1976 30: 201–216

    CAS  Google Scholar 

  16. Morris R, Argyris TS . Epidermal cell cycle and transit times during hyperplastic growth induced by abrasion or treatment with 12-O-tetradecanoylphorbol-13-acetate Cancer Res 1983 43: 4935–4942

    CAS  PubMed  Google Scholar 

  17. Yee JK et al. A general method for the generation of high-titer, pantropic retroviral vectors: highly efficient infection of primary hepatocytes Proc Natl Acad Sci USA 1994 91: 9564–9568

    Article  CAS  Google Scholar 

  18. Burns JC et al. Vesicular somatitis virus G glycoprotein pseudotyped retroviral vectors: concentration to very high titer and efficient gene transfer into mammalian and nonmammalian cells Proc Natl Acad Sci USA 1993 90: 8033–8037

    Article  CAS  Google Scholar 

  19. Fenjves ES, Yao S-N, Kurachi K, Taichman LB . Analysis of loss of expression of a retrovirus-transduced gene in grafted human keratinocytes J Invest Dermatol 1996 106: 576–578

    Article  CAS  Google Scholar 

  20. Choate KA, Medalie DA, Morgan JR, Khavari PA . Corrective gene transfer in the human skin disorder lamellar ichthyosis Nature 1996 2: 1263–1267

    CAS  Google Scholar 

  21. Gerrard AJ, Hudson DL, Brownlee GG, Watt FM . Towards gene therapy for haemophilia B using primary human keratinocytes Nature 1993 3: 180–183

    CAS  Google Scholar 

  22. White SJ, Page SM, Margaritis P, Brownlee GG . Long-term expression of human clotting factor IX from retrovirally transduced primary human keratinocytes in vivo Hum Gene Ther 1998 9: 1187–1195

    Article  CAS  Google Scholar 

  23. Rhim JA et al. Replacement of diseased mouse liver by hepatic cell transplantation Science 1994 263: 1149–1152

    Article  CAS  Google Scholar 

  24. Christiano AM, Uitto J . Molecular diagnosis of inherited skin diseases: the paradigm of dystrophic epidermolysis bullosa Adv Dermatol 1996 11: 199–213

    CAS  PubMed  Google Scholar 

  25. Dellambra E et al. Corrective transduction of human epidermal stem cells in laminin-5-dependent junctional epidermolysis bullosa Hum Gene Ther 1998 9: 1359–1370

    Article  CAS  Google Scholar 

  26. Garlick JA, Katz AB, Fenjves ES, Taichman LB . Retrovirus-mediated transduction of cultured epidermal keratinocytes J Invest Dermatol 1991 97: 824–829

    Article  CAS  Google Scholar 

  27. Kolodka TM, Garlick JA, Taichman LB . Evidence for keratinocyte stem cells in vitro: long term engraftment and persistence of transgene expression from retrovirus-transduced keratinocytes Proc Natl Acad Sci USA 1998 95: 4356–4361

    Article  CAS  Google Scholar 

  28. Mathor MB et al. Clonal analysis of stably transduced human epidermal stem cells in culture Proc Natl Acad Sci USA 1996 93: 10371–10376

    Article  CAS  Google Scholar 

  29. Hall PA, Watt FM . Stem cells: the generation and maintenance of cellular diversity Development 1989 106: 619–633

    CAS  PubMed  Google Scholar 

  30. Argyris TS . Kinetics of epidermal production during epidermal regeneration following abrasion in mice Am J Pathol 1976 83: 329–340

    CAS  PubMed  PubMed Central  Google Scholar 

  31. Lavker RM et al. Hair follicle stem cells: their location, role in hair cycle, and involvement in skin tumor formation J Invest Dermatol 1993 101: 16S–26S

    Article  CAS  Google Scholar 

  32. Kamimura J et al. Primary mouse keratinocytes cultures contain hair follicle progenitor cells with multiple differentation potential J Invest Dermatol 1997 109: 534–540

    Article  CAS  Google Scholar 

  33. Tuting T, Storkus WJ, Falo LD Jr . DNA immunization targeting the skin: molecular control of adaptive immunity J Invest Dermatol 1998 111: 183–188

    Article  CAS  Google Scholar 

  34. Raz E et al. Intradermal gene immunization: the possible role of DNA uptake in the induction of celluar immunity to viruses Proc Natl Acad Sci USA 1994 91: 9519–9523

    Article  CAS  Google Scholar 

  35. Steinman RM . The dendritic cell system and its role in immunogenicity Annu Rev Immunol 1991 9: 271–296

    Article  CAS  Google Scholar 

  36. Fava RA, McKanna J, Cohen S . Lipocortin I is abundant in a restricted number of differentiated cell types in adult organs J Cell Physiol 1989 141: 284–293

    Article  CAS  Google Scholar 

  37. Naldini L et al. In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector Science 1996 272: 263–267

    Article  CAS  Google Scholar 

  38. Ghazizadeh S, Harrington R, Garfield J, Taichman LB . Retrovirus-mediated transduction of porcine keratinocytes in organ culture J Invest Dermatol 1998 111: 492–496

    Article  CAS  Google Scholar 

  39. Kozarsky KF, Wilson JM . Gene therapy: adenovirus vectors Curr Opin Genet Dev 1993 3: 499–503

    Article  CAS  Google Scholar 

  40. Richardson C, Bank A . Developmental stage-specific expression and regulation of an amphotropic retroviral receptor in hematopoietic cells Mol Cell Biol 1996 16: 4240–4247

    Article  CAS  Google Scholar 

  41. Sambrook J, Fritsch EF, Maniatis T . Molecular Cloning: A Laboratory Manual Cold Spring Harbor Laboratory Press: Cold Spring Harbor 1989 pp 39–40

    Google Scholar 

Download references

Acknowledgements

We thank Drs Marcia Simon and Jonathan Garlick for helpful discussions, Chiron Technologies for the 293GP/LZRN cells, Dr Theodore Friedmann for the pHCMV-G plasmid, Drs Michael Blaese and Harry Muslow for the anti-NPTII antisera. This research was supported by grants from NIH (R37-DEO4511 and RO1-DK49093) and an NIH fellowship to SG (F32-AR08390).

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Authors and Affiliations

  1. Department of Oral Biology and Pathology, SUNY at Stony Brook, NY, USA

    S Ghazizadeh, R Harrington & L B Taichman

  2. Department of Dermatology, SUNY at Stony Brook, NY, USA

    L B Taichman

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  1. S Ghazizadeh
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  2. R Harrington
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  3. L B Taichman
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Ghazizadeh, S., Harrington, R. & Taichman, L. In vivo transduction of mouse epidermis with recombinant retroviral vectors: implications for cutaneous gene therapy. Gene Ther 6, 1267–1275 (1999). https://doi.org/10.1038/sj.gt.3300956

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