Download PDF
Thromb Haemost 1999; 81(02): 234-239
DOI: 10.1055/s-0037-1614449
Review Articles
Schattauer GmbH

Evaluation of an Adenoviral Vector Encoding Full-Length Human Factor VIII in Hemophiliac Mice

Sheila Connelly
3   Genetic Therapy, Inc. (A Novartis Company), Gaithersburg, MD, USA
,
Julie L. Andrews
3   Genetic Therapy, Inc. (A Novartis Company), Gaithersburg, MD, USA
,
Angela M. Gallo-Penn
3   Genetic Therapy, Inc. (A Novartis Company), Gaithersburg, MD, USA
,
Luigina Tagliavacca
1   From the Department of Biological Chemistry, Gaithersburg, MD, USA
,
Randal J. Kaufman
2   Howard Hughes Medical Institute, University of Michigan Medical School Ann Arbor, MI, Gaithersburg, MD, USA
,
Michael Kaleko
3   Genetic Therapy, Inc. (A Novartis Company), Gaithersburg, MD, USA
› Author Affiliations
Further Information

Publication History

Received26 June 1998

Accepted after resubmission26 October 1998

Publication Date:
08 December 2017 (online)

    Permissions and Reprints

Summary

Adenoviral vectors provide a promising gene therapy system for the treatment of hemophilia A. Potent vectors encoding a human factor VIII (FVIII) cDNA were developed that mediated sustained FVIII expression in normal and hemophiliac mice and complete phenotypic correction of the bleeding disorder in hemophiliac mice and dogs (Connelly and Kaleko, Haemophilia 1998; 4: 380-8). However, these studies utilized vectors encoding a truncated version of the human FVIII cDNA lacking the B-domain (BDD FVIII). In this work, an adenoviral vector encoding the human full-length (FL) FVIII cDNA was generated and characterized. While functional FL FVIII was secreted in vitro, expression of the FL protein was not detected in the plasma of vector-treated hemophiliac mice. Unexpectedly, the FL FVIII vector-treated animals demonstrated phenotypic correction of the bleeding defect as measured by a tail-clip survival study. FL FVIII protein was visualized in the mouse livers using human FVIII-specific immunohistochemical analyses. These data demonstrate that adenoviral vector-mediated in vivo expression of BDD FVIII is more efficient than that of the FL protein and that phenotypic correction can occur in the absence of detectable levels of FVIII.

 

  • References

  • 1 Connelly S, Kaleko M. Gene therapy for hemophilia A. Thromb Haemost 1997; 78: 31-6.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 2 Connelly S, Kaleko M. Haemophilia A gene therapy. Haemophilia 1998; 4: 380-8.
    CrossrefPubMedGoogle Scholar
  • 3 Smith TAG. Gene therapy for haemophilia. Exp Opin Invest Drugs 1997; 6: 1685-90.
    CrossrefPubMedGoogle Scholar
  • 4 Connelly S, Smith TAG, Dhir G, Gardner JM, Mehaffey MG, Zaret KS, McClelland A, Kaleko M. In vivo gene delivery and expression of physiological levels of functional human factor VIII in mice. Hum Gene Ther 1995; 6: 185-93.
    CrossrefPubMedGoogle Scholar
  • 5 Connelly S, Gardner JM, Lyons RM, McClelland A, Kaleko M. Sustained expression of therapeutic levels of human factor VIII in mice. Blood 1996; 87: 4671-7.
    PubMedGoogle Scholar
  • 6 Connelly S, Gardner JM, McClelland A, Kaleko M. High level tissue-specific expression of functional human factor VIII in mice. Hum Gene Ther 1996; 7: 183-95.
    CrossrefPubMedGoogle Scholar
  • 7 Connelly S, Andrews J, Gallo AM, Qian J, Hoyer L, Kadan MJ, Gorziglia MI, Trapnell BC, McClelland A, Kaleko M. Sustained phenotypic correction of murine hemophilia A by in vivo gene therapy. Blood 1998; 91: 3273-81.
    PubMedGoogle Scholar
  • 8 Connelly S, Mount J, Mauser A, Gardner JM, Kaleko M, McClelland A, Lothrop CD. Complete short-term correction of canine hemophilia A by in vivo gene therapy. Blood 1996; 88: 3846-53.
    PubMedGoogle Scholar
  • 9 Berkner KL. Development of adenovirus vectors for the expression of heterologous genes. Biotechniques 1988; 6: 616-29.
    PubMedGoogle Scholar
  • 10 Gorziglia I M, Kadan MJ, Yei S, Lim J, Lee GM, Luthra R, Trapnell BC. Elimination of both E1 and E2a from adenovirus vectors further improves prospects for in vivo human gene therapy. J Virol 1996; 6: 4173-8.
    PubMedGoogle Scholar
  • 11 Engelhardt JF, Ye X, Doranz B, Wilson JM. Ablation of E2A in recombinant adenoviruses improves transgene persistence and decreases inflammatory response in mouse liver. Proc Natl Acad Sci USA 1994; 91: 6196-200.
    CrossrefPubMedGoogle Scholar
  • 12 Engelhardt JF, Litzky L, Wilson JM. Prolonged transgene expression in cotton rat lung with recombinant adenoviruses defective in E2a. Hum Gene Ther 1994; 5: 1217-29.
    CrossrefPubMedGoogle Scholar
  • 13 Yang Y, Nunes FA, Berencsi K, Gonczol E, Engelhardt JF, Wilson JM. Inactivation of E2a in recombinant adenoviruses improves the prospect for gene therapy in cystic fibrosis. Nat Genet 1994; 7: 362-9.
    CrossrefPubMedGoogle Scholar
  • 14 Morral N, O’Neal W, Zhou H, Langston C, Beaudet A. Immune responses to reporter proteins and high viral dose limit duration of expression with adenoviral vectors: comparison of E2a wild type and E2a deleted vectors. Hum Gene Ther 1997; 8: 1275-86.
    CrossrefPubMedGoogle Scholar
  • 15 Fang B, Wang H, Gordon G, Bellinger DA, Read MS, Brinkhous KM, Woo SL, Eisensmith RC. Lack of persistence of E1- recombinant adenoviral vectors containing a temperature-sensitive E2A mutation in immunocompetent mice and hemophilia B dogs. Gene Therapy 1996; 3: 217-22.
    PubMedGoogle Scholar
  • 16 Eaton DL, Wood WI, Eaton D, Hass PE, Hollingshead P, Wion K, Mather J, Lawn RM, Vehar GA, Gorman C. Construction and characterization of an active factor VIII variant lacking the central one-third of the molecule. Biochem 1986; 25: 1-2.
    CrossrefPubMedGoogle Scholar
  • 17 Toole JJ, Pittman DD, Orr EC, Murtha P, Wasley LC, Kaufman RJ. A large region (~95 kDa of human factor VIII is dispensable for in vitro procoagulant activity. Proc Natl Acad Sci USA 1986; 83: 5939-42.
    CrossrefPubMedGoogle Scholar
  • 18 Meulien P, Faure T, Mischler F, Harrer H, Ulrich P, Bouderbala B, Dott K, Sainte Marie M, Mazurier C, Wiesel ML. A new recombinant procoagulant protein derived from the cDNA encoding human factor VIII. Protein Eng 1988; 2: 301-6.
    CrossrefPubMedGoogle Scholar
  • 19 Sarver N, Ricca GA, Link J, Nathan MH, Newman J, Drohan WN. Stable expression of recombinant factor VIII molecules using a bovine papillomavirus vector. DNA 1987; 6: 553-64.
    CrossrefPubMedGoogle Scholar
  • 20 Pittman DD, Alderman EM, Tomkinson KN, Wang JH, Giles AR, Kaufman RJ. Biochemical, Immunological, and in vivo functional characterization of B-domain-deleted factor VIII. Blood 1993; 81: 2925-35.
    PubMedGoogle Scholar
  • 21 Dorner AJ, Bole DG, Kaufman RJ. The relationship of N-linked glycosylation and heavy chain-binding protein association with the secretion of glycoproteins. J Cell Biol 1987; 105: 2665-74.
    CrossrefPubMedGoogle Scholar
  • 22 Dorner AJ, Wasley LC, Kaufman RJ. Overexpression of GRP78 mitigates stress induction of glucose regulated proteins and blocks secretion of selective proteins in Chinese hamster ovary cells. EMBO J 1992; 11: 1563-71.
    PubMedGoogle Scholar
  • 23 Mittereder N, March KL, Trapnell BC. Evaluation of the concentration and bioactivity of adenovirus vectors for gene therapy. J Virol 1996; 70: 7498-509.
    PubMedGoogle Scholar
  • 24 Tolstoshev P, Mittereder N, Yei S, Trapnell BC. Adenovirus vectors for gene therapy. In: Cancer chemotherapy: Challenges for the future. Excerpta Medica, Ltd; Tokyo: 1994: pp. 23-31.
    Google Scholar
  • 25 Knowles BB, Howe CC, Aden DP. Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science 1980; 209: 497-9.
    CrossrefPubMedGoogle Scholar
  • 26 Bi L, Lawler AM, Antonarakis SE, High KA, Gerhart JD, Kazazian HH. Targeted disruption of the mouse factor VIII gene produces a model of haemophilia A. Nature Genetics 1995; 10: 119-21.
    CrossrefPubMedGoogle Scholar
  • 27 Kay MA, Rothenberg SR, Landen CN, Bellinger DA, Leland F, Toman C, Finegold M, Thompson AR, Read MS, Brinkhous KM, Woo SLC. In vivo gene therapy of hemophilia B: Sustained partial correction in factor IX-deficient dogs. Science 1993; 262: 117-9.
    CrossrefPubMedGoogle Scholar
  • 28 Berntorp E. Second generation, B-domain deleted recombinant factor VIII. Thromb Haemost 1997; 78: 256-60.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 29 Fijnvandraat K, Berntorp E, ten Cate JW, Johnsson H, Peters M, Savidge G, Tengborn L, Spira J, Stahl C. Recombinant, B-domain deleted factor VIII (r-VIII SQ): pharmacokinetics and initial safety aspects in hemophilia A patients. Thromb Haemost 1997; 77: 298-302.
    Article in Thieme ConnectPubMedGoogle Scholar
  • 30 Rosenberg JB, Foster PA, Kaufman RJ, Vokac EA, Moussalli M, Kroner PA, Montgomery RR. Intracellular trafficking of factor VIII to von Willebrand factor storage granules. J Clin Invest 1998; 101: 613-24.
    CrossrefPubMedGoogle Scholar
  • 31 Swaroop M, Moussalli M, Pipe SW, Kaufman RJ. Mutagenesis of a potential immunoglobulin-binding protein-binding site enhances secretion of coagulation factor VIII. J Biol Chem 1997; 272: 24121-4.
    CrossrefPubMedGoogle Scholar
  • 32 Stel HV, van der Kwast TH, Veerman EC. Detection of factor VIII/coagulant antigen in human liver tissue. Nature 1983; 303: 530-2.
    CrossrefPubMedGoogle Scholar
  • 33 Stel HV, Oudejan CB, Veerman EC, Meijer CJ. Immunohistological localization of factor VIII in placental endothelial cells. Br J Haematol 1986; 63: 565-9.
    CrossrefPubMedGoogle Scholar
  • 34 Schiedner G, Morral N, Parks RJ, Wu Y, Koopmans SC, Langston C, Graham FL, Beaudet AL, Kochanek S. Genomic DNA transfer with a high-capacity adenovirus vector results in improved in vivo gene expression and decreased toxicity. Nat Genet 1998; 18: 180-3.
    CrossrefPubMedGoogle Scholar
  • 35 Morsy MA, Gu M, Motzel S, Zhau J, Lin J, Su Q, Allen H, Franlin L, Parks RJ, Graham FL, Kochanek S, Bett AJ, Caskey CT. An adenoviral vector deleted for all viral coding sequences results in enhanced safety and extended expression of a leptin transgene. Proc Natl Acad Sci USA 1998; 95: 7866-71.
    CrossrefPubMedGoogle Scholar
  • 36 Hoyer LW. Hemophilia A. N Engl J Med 1994; 330: 38-47.
    CrossrefPubMedGoogle Scholar