Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Dec 6;91(25):12188–12192. doi: 10.1073/pnas.91.25.12188

In vivo transfer and expression of a human epidermal growth factor gene accelerates wound repair.

C Andree 1, W F Swain 1, C P Page 1, M D Macklin 1, J Slama 1, D Hatzis 1, E Eriksson 1
PMCID: PMC45402  PMID: 7991604

Abstract

This report details the transfer of a human epidermal growth factor (hEGF) expression plasmid to porcine partial-thickness wound keratinocytes by particle-mediated DNA transfer (Accell). After gene transfer an external sealed fluid-filled wound chamber was used to protect the wound, provide containment of the exogenous DNA and expressed peptide, and permit sampling of the wound fluid. Analysis of wound fluid for hEGF and total protein, an indicator of reformation of the epithelial barrier, showed that wounds bombarded with the hEGF plasmid exhibited a 190-fold increase in EGF concentration and healed 20% (2.1 days) earlier than the controls. EGF concentrations in wound fluid persisted over the entire 10-day monitored period, decreasing from 200 pg/ml to 25 pg/ml over the first 5 days. Polymerase chain reaction results showed that plasmid DNA was present in the wound for at least 30 days. These findings demonstrate the possible utility of in vivo gene transfer to enhance epidermal repair.

Full text

PDF
12192

Images in this article

12190Image
on p.12190
12191Image
on p.12191
12191Image
on p.12191

Selected References

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

  1. Barr E., Leiden J. M. Systemic delivery of recombinant proteins by genetically modified myoblasts. Science. 1991 Dec 6;254(5037):1507–1509. doi: 10.1126/science.1962212. [DOI] [PubMed] [Google Scholar]
  2. Barrandon Y., Morgan J. R., Mulligan R. C., Green H. Restoration of growth potential in paraclones of human keratinocytes by a viral oncogene. Proc Natl Acad Sci U S A. 1989 Jun;86(11):4102–4106. doi: 10.1073/pnas.86.11.4102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bell G. I., Fong N. M., Stempien M. M., Wormsted M. A., Caput D., Ku L. L., Urdea M. S., Rall L. B., Sanchez-Pescador R. Human epidermal growth factor precursor: cDNA sequence, expression in vitro and gene organization. Nucleic Acids Res. 1986 Nov 11;14(21):8427–8446. doi: 10.1093/nar/14.21.8427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boshart M., Weber F., Jahn G., Dorsch-Häsler K., Fleckenstein B., Schaffner W. A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus. Cell. 1985 Jun;41(2):521–530. doi: 10.1016/s0092-8674(85)80025-8. [DOI] [PubMed] [Google Scholar]
  5. Breuing K., Eriksson E., Liu P., Miller D. R. Healing of partial thickness porcine skin wounds in a liquid environment. J Surg Res. 1992 Jan;52(1):50–58. doi: 10.1016/0022-4804(92)90278-8. [DOI] [PubMed] [Google Scholar]
  6. Brown G. L., Curtsinger L., 3rd, Brightwell J. R., Ackerman D. M., Tobin G. R., Polk H. C., Jr, George-Nascimento C., Valenzuela P., Schultz G. S. Enhancement of epidermal regeneration by biosynthetic epidermal growth factor. J Exp Med. 1986 May 1;163(5):1319–1324. doi: 10.1084/jem.163.5.1319. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Brown G. L., Curtsinger L., Jurkiewicz M. J., Nahai F., Schultz G. Stimulation of healing of chronic wounds by epidermal growth factor. Plast Reconstr Surg. 1991 Aug;88(2):189–196. [PubMed] [Google Scholar]
  8. Brown G. L., Nanney L. B., Griffen J., Cramer A. B., Yancey J. M., Curtsinger L. J., 3rd, Holtzin L., Schultz G. S., Jurkiewicz M. J., Lynch J. B. Enhancement of wound healing by topical treatment with epidermal growth factor. N Engl J Med. 1989 Jul 13;321(2):76–79. doi: 10.1056/NEJM198907133210203. [DOI] [PubMed] [Google Scholar]
  9. Buckley A., Davidson J. M., Kamerath C. D., Wolt T. B., Woodward S. C. Sustained release of epidermal growth factor accelerates wound repair. Proc Natl Acad Sci U S A. 1985 Nov;82(21):7340–7344. doi: 10.1073/pnas.82.21.7340. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Buckley A., Davidson J. M., Kamerath C. D., Woodward S. C. Epidermal growth factor increases granulation tissue formation dose dependently. J Surg Res. 1987 Oct;43(4):322–328. doi: 10.1016/0022-4804(87)90088-6. [DOI] [PubMed] [Google Scholar]
  11. COHEN S. Isolation of a mouse submaxillary gland protein accelerating incisor eruption and eyelid opening in the new-born animal. J Biol Chem. 1962 May;237:1555–1562. [PubMed] [Google Scholar]
  12. Chowdhury J. R., Grossman M., Gupta S., Chowdhury N. R., Baker J. R., Jr, Wilson J. M. Long-term improvement of hypercholesterolemia after ex vivo gene therapy in LDLR-deficient rabbits. Science. 1991 Dec 20;254(5039):1802–1805. doi: 10.1126/science.1722351. [DOI] [PubMed] [Google Scholar]
  13. Cohen S. Epidermal growth factor. J Invest Dermatol. 1972 Jul;59(1):13–16. doi: 10.1111/1523-1747.ep12625690. [DOI] [PubMed] [Google Scholar]
  14. Cornetta K., Morgan R. A., Anderson W. F. Safety issues related to retroviral-mediated gene transfer in humans. Hum Gene Ther. 1991 Spring;2(1):5–14. doi: 10.1089/hum.1991.2.1-5. [DOI] [PubMed] [Google Scholar]
  15. DeNoto F. M., Moore D. D., Goodman H. M. Human growth hormone DNA sequence and mRNA structure: possible alternative splicing. Nucleic Acids Res. 1981 Aug 11;9(15):3719–3730. doi: 10.1093/nar/9.15.3719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Dhawan J., Pan L. C., Pavlath G. K., Travis M. A., Lanctot A. M., Blau H. M. Systemic delivery of human growth hormone by injection of genetically engineered myoblasts. Science. 1991 Dec 6;254(5037):1509–1512. doi: 10.1126/science.1962213. [DOI] [PubMed] [Google Scholar]
  17. Falanga V., Eaglstein W. H., Bucalo B., Katz M. H., Harris B., Carson P. Topical use of human recombinant epidermal growth factor (h-EGF) in venous ulcers. J Dermatol Surg Oncol. 1992 Jul;18(7):604–606. doi: 10.1111/j.1524-4725.1992.tb03514.x. [DOI] [PubMed] [Google Scholar]
  18. Flowers M. E., Stockschlaeder M. A., Schuening F. G., Niederwieser D., Hackman R., Miller A. D., Storb R. Long-term transplantation of canine keratinocytes made resistant to G418 through retrovirus-mediated gene transfer. Proc Natl Acad Sci U S A. 1990 Mar;87(6):2349–2353. doi: 10.1073/pnas.87.6.2349. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. MacGregor G. R., Caskey C. T. Construction of plasmids that express E. coli beta-galactosidase in mammalian cells. Nucleic Acids Res. 1989 Mar 25;17(6):2365–2365. doi: 10.1093/nar/17.6.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. MacGregor G. R., Mogg A. E., Burke J. F., Caskey C. T. Histochemical staining of clonal mammalian cell lines expressing E. coli beta galactosidase indicates heterogeneous expression of the bacterial gene. Somat Cell Mol Genet. 1987 May;13(3):253–265. doi: 10.1007/BF01535207. [DOI] [PubMed] [Google Scholar]
  21. Marikovsky M., Breuing K., Liu P. Y., Eriksson E., Higashiyama S., Farber P., Abraham J., Klagsbrun M. Appearance of heparin-binding EGF-like growth factor in wound fluid as a response to injury. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3889–3893. doi: 10.1073/pnas.90.9.3889. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Morgan J. R., Barrandon Y., Green H., Mulligan R. C. Expression of an exogenous growth hormone gene by transplantable human epidermal cells. Science. 1987 Sep 18;237(4821):1476–1479. doi: 10.1126/science.3629250. [DOI] [PubMed] [Google Scholar]
  23. Mulligan R. C. The basic science of gene therapy. Science. 1993 May 14;260(5110):926–932. doi: 10.1126/science.8493530. [DOI] [PubMed] [Google Scholar]
  24. Mullis K. B., Faloona F. A. Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction. Methods Enzymol. 1987;155:335–350. doi: 10.1016/0076-6879(87)55023-6. [DOI] [PubMed] [Google Scholar]
  25. Ponder K. P., Gupta S., Leland F., Darlington G., Finegold M., DeMayo J., Ledley F. D., Chowdhury J. R., Woo S. L. Mouse hepatocytes migrate to liver parenchyma and function indefinitely after intrasplenic transplantation. Proc Natl Acad Sci U S A. 1991 Feb 15;88(4):1217–1221. doi: 10.1073/pnas.88.4.1217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Read L. C., George-Nascimento C. Epidermal growth factor: physiological roles and therapeutic applications. Biotechnol Ther. 1989 1990;1(3):237–272. [PubMed] [Google Scholar]
  27. Schultz G. S., White M., Mitchell R., Brown G., Lynch J., Twardzik D. R., Todaro G. J. Epithelial wound healing enhanced by transforming growth factor-alpha and vaccinia growth factor. Science. 1987 Jan 16;235(4786):350–352. doi: 10.1126/science.3492044. [DOI] [PubMed] [Google Scholar]
  28. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  29. Temin H. M. Safety considerations in somatic gene therapy of human disease with retrovirus vectors. Hum Gene Ther. 1990 Summer;1(2):111–123. doi: 10.1089/hum.1990.1.2-111. [DOI] [PubMed] [Google Scholar]
  30. Teumer J., Lindahl A., Green H. Human growth hormone in the blood of athymic mice grafted with cultures of hormone-secreting human keratinocytes. FASEB J. 1990 Nov;4(14):3245–3250. doi: 10.1096/fasebj.4.14.2227214. [DOI] [PubMed] [Google Scholar]
  31. Vogt P. M., Thompson S., Andree C., Liu P., Breuing K., Hatzis D., Brown H., Mulligan R. C., Eriksson E. Genetically modified keratinocytes transplanted to wounds reconstitute the epidermis. Proc Natl Acad Sci U S A. 1994 Sep 27;91(20):9307–9311. doi: 10.1073/pnas.91.20.9307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Yang N. S., Burkholder J., Roberts B., Martinell B., McCabe D. In vivo and in vitro gene transfer to mammalian somatic cells by particle bombardment. Proc Natl Acad Sci U S A. 1990 Dec;87(24):9568–9572. doi: 10.1073/pnas.87.24.9568. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES