Skip to main content
NCBI 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 May 24;91(11):5148–5152. doi: 10.1073/pnas.91.11.5148

In vivo production of human factor VII in mice after intrasplenic implantation of primary fibroblasts transfected by receptor-mediated, adenovirus-augmented gene delivery.

K Zatloukal 1, M Cotten 1, M Berger 1, W Schmidt 1, E Wagner 1, M L Birnstiel 1
PMCID: PMC43949  PMID: 8197198

Abstract

Hemophilia A is caused by defects in the factor VIII gene. This results in life-threatening hemorrhages and severe arthropathies. Today, hemophiliacs are treated with human blood-derived factor VIII. In the future, it may be possible to use gene therapy to avoid long-term complications of conventional therapy and to improve the quality of life. However, initial gene therapy models using retroviral vectors and nonviral gene transfer techniques to introduce factor VIII gene constructs have been hampered by low expression levels of factor VIII. We show here that high expression levels of the B-domain-deleted human factor VIII in primary mouse fibroblasts and myoblasts are obtained by using receptor-mediated, adenovirus-augmented gene delivery (transferrinfection). We demonstrate that, presumably owing to the high molecular weight of factor VIII or its metabolic instability, secretion into the blood and attainment of therapeutic in vivo levels of factor VIII is achieved only if transfected autologous primary fibroblasts or myoblasts are delivered to the liver or spleen, but not if myoblasts are implanted into muscle, a strategy known to be successful for factor IX delivery.

Full text

PDF
5148

Images in this article

5150Image
on p.5150
5150Image
on p.5150

Selected References

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

  1. Bontempo F. A., Lewis J. H., Gorenc T. J., Spero J. A., Ragni M. V., Scott J. P., Starzl T. E. Liver transplantation in hemophilia A. Blood. 1987 Jun;69(6):1721–1724. [PMC free article] [PubMed] [Google Scholar]
  2. Cotten M., Wagner E., Zatloukal K., Phillips S., Curiel D. T., Birnstiel M. L. High-efficiency receptor-mediated delivery of small and large (48 kilobase gene constructs using the endosome-disruption activity of defective or chemically inactivated adenovirus particles. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6094–6098. doi: 10.1073/pnas.89.13.6094. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Curiel D. T., Wagner E., Cotten M., Birnstiel M. L., Agarwal S., Li C. M., Loechel S., Hu P. C. High-efficiency gene transfer mediated by adenovirus coupled to DNA-polylysine complexes. Hum Gene Ther. 1992 Apr;3(2):147–154. doi: 10.1089/hum.1992.3.2-147. [DOI] [PubMed] [Google Scholar]
  4. Dai Y., Roman M., Naviaux R. K., Verma I. M. Gene therapy via primary myoblasts: long-term expression of factor IX protein following transplantation in vivo. Proc Natl Acad Sci U S A. 1992 Nov 15;89(22):10892–10895. doi: 10.1073/pnas.89.22.10892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eaton D. L., Wood W. I., Eaton D., Hass P. E., Hollingshead P., Wion K., Mather J., Lawn R. M., Vehar G. A., Gorman C. Construction and characterization of an active factor VIII variant lacking the central one-third of the molecule. Biochemistry. 1986 Dec 30;25(26):8343–8347. doi: 10.1021/bi00374a001. [DOI] [PubMed] [Google Scholar]
  6. Hoeben R. C., Einerhand M. P., Briët E., van Ormondt H., Valerio D., van der Eb A. J. Toward gene therapy in haemophilia A: retrovirus-mediated transfer of a factor VIII gene into murine haematopoietic progenitor cells. Thromb Haemost. 1992 Mar 2;67(3):341–345. [PubMed] [Google Scholar]
  7. Hoeben R. C., Fallaux F. J., Van Tilburg N. H., Cramer S. J., Van Ormondt H., Briët E., Van Der Eb A. J. Toward gene therapy for hemophilia A: long-term persistence of factor VIII-secreting fibroblasts after transplantation into immunodeficient mice. Hum Gene Ther. 1993 Apr;4(2):179–186. doi: 10.1089/hum.1993.4.2-179. [DOI] [PubMed] [Google Scholar]
  8. Hoeben R. C., van der Jagt R. C., Schoute F., van Tilburg N. H., Verbeet M. P., Briët E., van Ormondt H., van der Eb A. J. Expression of functional factor VIII in primary human skin fibroblasts after retrovirus-mediated gene transfer. J Biol Chem. 1990 May 5;265(13):7318–7323. [PubMed] [Google Scholar]
  9. Israel D. I., Kaufman R. J. Retroviral-mediated transfer and amplification of a functional human factor VIII gene. Blood. 1990 Mar 1;75(5):1074–1080. [PubMed] [Google Scholar]
  10. Kaufman R. J. Developing rDNA products for treatment of hemophilia A. Trends Biotechnol. 1991 Oct;9(10):353–359. doi: 10.1016/0167-7799(91)90118-2. [DOI] [PubMed] [Google Scholar]
  11. Lewis J. H., Bontempo F. A., Spero J. A., Ragni M. V., Starzl T. E. Liver transplantation in a hemophiliac. N Engl J Med. 1985 May 2;312(18):1189–1190. doi: 10.1056/NEJM198505023121812. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lynch C. M., Israel D. I., Kaufman R. J., Miller A. D. Sequences in the coding region of clotting factor VIII act as dominant inhibitors of RNA accumulation and protein production. Hum Gene Ther. 1993 Jun;4(3):259–272. doi: 10.1089/hum.1993.4.3-259. [DOI] [PubMed] [Google Scholar]
  13. Marchioro T. L., Hougie C., Ragde H., Epstein R. B., Thomas E. D. Hemophilia: role of organ homografts. Science. 1969 Jan 10;163(3863):188–190. doi: 10.1126/science.163.3863.188. [DOI] [PubMed] [Google Scholar]
  14. 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]
  15. Powell J. T., Klaasse Bos J. M., van Mourik J. A. The uptake and expression of the factor VIII and reporter genes by vascular cells. FEBS Lett. 1992 Jun 1;303(2-3):173–177. doi: 10.1016/0014-5793(92)80512-f. [DOI] [PubMed] [Google Scholar]
  16. Roman M., Axelrod J. H., Dai Y., Naviaux R. K., Friedmann T., Verma I. M. Circulating human or canine factor IX from retrovirally transduced primary myoblasts and established myoblast cell lines grafted into murine skeletal muscle. Somat Cell Mol Genet. 1992 May;18(3):247–258. doi: 10.1007/BF01233861. [DOI] [PubMed] [Google Scholar]
  17. Thompson A. R. Status of gene transfer for hemophilia A and B. Thromb Haemost. 1991 Jul 12;66(1):119–122. [PubMed] [Google Scholar]
  18. Toole J. J., Knopf J. L., Wozney J. M., Sultzman L. A., Buecker J. L., Pittman D. D., Kaufman R. J., Brown E., Shoemaker C., Orr E. C. Molecular cloning of a cDNA encoding human antihaemophilic factor. Nature. 1984 Nov 22;312(5992):342–347. doi: 10.1038/312342a0. [DOI] [PubMed] [Google Scholar]
  19. Toole J. J., Pittman D. D., Orr E. C., Murtha P., Wasley L. C., Kaufman R. J. A large region (approximately equal to 95 kDa) of human factor VIII is dispensable for in vitro procoagulant activity. Proc Natl Acad Sci U S A. 1986 Aug;83(16):5939–5942. doi: 10.1073/pnas.83.16.5939. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Vehar G. A., Keyt B., Eaton D., Rodriguez H., O'Brien D. P., Rotblat F., Oppermann H., Keck R., Wood W. I., Harkins R. N. Structure of human factor VIII. Nature. 1984 Nov 22;312(5992):337–342. doi: 10.1038/312337a0. [DOI] [PubMed] [Google Scholar]
  21. Wagner E., Zatloukal K., Cotten M., Kirlappos H., Mechtler K., Curiel D. T., Birnstiel M. L. Coupling of adenovirus to transferrin-polylysine/DNA complexes greatly enhances receptor-mediated gene delivery and expression of transfected genes. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):6099–6103. doi: 10.1073/pnas.89.13.6099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wion K. L., Kelly D., Summerfield J. A., Tuddenham E. G., Lawn R. M. Distribution of factor VIII mRNA and antigen in human liver and other tissues. Nature. 1985 Oct 24;317(6039):726–729. doi: 10.1038/317726a0. [DOI] [PubMed] [Google Scholar]
  23. Wood W. I., Capon D. J., Simonsen C. C., Eaton D. L., Gitschier J., Keyt B., Seeburg P. H., Smith D. H., Hollingshead P., Wion K. L. Expression of active human factor VIII from recombinant DNA clones. Nature. 1984 Nov 22;312(5992):330–337. doi: 10.1038/312330a0. [DOI] [PubMed] [Google Scholar]
  24. Yao S. N., Kurachi K. Expression of human factor IX in mice after injection of genetically modified myoblasts. Proc Natl Acad Sci U S A. 1992 Apr 15;89(8):3357–3361. doi: 10.1073/pnas.89.8.3357. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Zatloukal K., Wagner E., Cotten M., Phillips S., Plank C., Steinlein P., Curiel D. T., Birnstiel M. L. Transferrinfection: a highly efficient way to express gene constructs in eukaryotic cells. Ann N Y Acad Sci. 1992 Oct 28;660:136–153. doi: 10.1111/j.1749-6632.1992.tb21066.x. [DOI] [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