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Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer

Gene Therapy volume 6, pages 1249–1257 (1999)Cite this article

Abstract

Developmental patterns of gene expression were determined following intravascular administration of adenovirus in utero, during sequential stages of murine development. Replication-deficient adenovirus (AdCMV.LacZ) was injected into yolk sac vessels of mouse embryos 12, 13, 15 and 18 days post-conception (d.p.c.). β-Galactosidase (β-gal) expression was evaluated 24–48 h after injection, at birth, and 5 weeks following normal delivery. Gene expression was detected in myocardial cells, endothelial cells of heart, lung, kidney, adrenal, gut, and in hepatocytes. The patterns of expression were distinct for each stage of virus administration and time-point of analysis. Intensity of individual organ expression varied with injection time-point, with the largest number of organs express- ing the transgene when embryos were injected at 15 d.p.c. β-Gal activity was detected in only a subset of cells expressing the murine coxsackievirus and adenovirus receptor (CAR), indicating factors other than receptor distribution were responsible for the pattern of transgene expression observed. These studies begin to define critical parameters affecting intravascular gene delivery in utero and indicate that intrinsic developmental regulatory mechanisms may control exogenous gene expression. Intravenous administration of adenovirus provides a unique approach for in utero gene transduction and will be a useful adjunct in evaluating genes which have early lethal mutations.

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References

  1. Copp AJ . Death before birth: clues from gene knockouts and mutations Trends Genet 1995 11: 87–93

    Article  CAS  PubMed  Google Scholar 

  2. Rossant J . Mouse mutants and cardiac development; new molecular insights into cardiogenesis Circ Res 1996 78: 349–353

    Article  CAS  PubMed  Google Scholar 

  3. Fishman GI . Timing is everything in life: conditional transgene expression in the cardiovascular system Circ Res 1998 82: 837–844

    Article  CAS  PubMed  Google Scholar 

  4. Pitt BR et al. Retrovirus-mediated gene transfer in lungs of living fetal sheep Gene Therapy 1995 2: 344–350

    CAS  PubMed  Google Scholar 

  5. McCray PB et al. Adenoviral-mediated gene transfer to fetal pulmonary epithelia in vitro and in vivo J Clin Invest 1995 95: 2620–2632

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Vincent MC et al. Adenovirus-mediated gene transfer to the respiratory tract of fetal sheep in utero Hum Gene Ther 1995 6: 1019–1028

    Article  CAS  PubMed  Google Scholar 

  7. Sekhon H, Larson JE . In utero gene transfer into the pulmonary epithelium Nature Med 1995 1: 1201–1203

    Article  CAS  PubMed  Google Scholar 

  8. Holzinger A et al. Intraamniotic administration of an adenoviral vector for gene transfer to fetal sheep and mouse tissues Pediat Res 1995 38: 844–850

    Article  CAS  PubMed  Google Scholar 

  9. Clapp DW, Dumenco LL, Hatzoglou M, Gerson SL . Fetal liver hematopoietic stem cells as a target for in utero retroviral gene transfer Blood 1991 78: 1132–1139

    CAS  PubMed  Google Scholar 

  10. Clapp DW et al. Myeloproliferative sarcoma virus directed expression of beta-galactosidase following retroviral transduction of murine hematopoietic cells Exp Hematol 1995 23: 630–638

    CAS  PubMed  Google Scholar 

  11. Wang G et al. Ultrasound-guided gene transfer to hepatocytes in utero Fetal Diagn Ther 1998 13: 197–205

    Article  PubMed  Google Scholar 

  12. Kass-Eisler A et al. The impact of developmental stage, route of administration and the immune system on adenovirus-meditated gene transfer Gene Therapy 1994 1: 395–402

    CAS  PubMed  Google Scholar 

  13. Tsukamoto M et al. Gene transfer and expression in progeny after intravenous DNA injection into pregnant mice Nat Genet 1995 9: 243–248

    Article  CAS  PubMed  Google Scholar 

  14. Kantoff PW et al. In utero gene transfer and expression: a sheep transplantation model Blood 1989 73: 1066–1073

    CAS  PubMed  Google Scholar 

  15. Hatzoglou M et al. Hepatic gene transfer in animals using retroviruses containing the promoter for the gene for phosphoenolpyruvate carboxykinase J Biol Chem 1990 265: 17285–17293

    CAS  PubMed  Google Scholar 

  16. Woo YJ et al. In utero cardiac gene transfer via intraplacental delivery of recombinant adenovirus Circulation 1997 96: 3561–3569

    Article  CAS  PubMed  Google Scholar 

  17. Fleischman RA, Mintz B . Prevention of genetic anemias in mice by microinjection of normal hematopoietic stem cells into the fetal placenta Proc Natl Acad Sci USA 1979 76: 5736–5740

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Senut MC, Suhr ST, Gage FH . Gene transfer to the rodent placenta in situ. A new strategy for delivering gene products to the fetus J Clin Invest 1998 101: 1565–1571

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Huard J et al. The route of administration is a major determinant of transduction efficiency of rat tissues by adenoviral recombinants Gene Therapy 1995 2: 107–115

    CAS  PubMed  Google Scholar 

  20. Stratford-Perricaudet LD, Makeh I, Perricaudet M, Briand P . Widespread long-term gene transfer to mouse skeletal muscles and heart J Clin Invest 1992 90: 626–630

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Hogers B et al. Intracardiac blood flow patterns related to the yolk sac circulation of the chick embryo Circ Res 1995 76: 871–877

    Article  CAS  PubMed  Google Scholar 

  22. Rasanen J et al. Role of the pulmonary circulation in the distribution of human fetal cardiac output during the second half of pregnancy Circulation 1996 94: 1068–1073

    Article  CAS  PubMed  Google Scholar 

  23. Bergelson JM et al. The murine CAR homolog is a receptor for coxsackie B viruses and adenoviruses J Virol 1998 72: 415–419

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Wickham TJ, Mathias P, Cheresh DA, Nemerow GR . Integrins alpha v beta 3 and alpha v beta 5 promote adenovirus internalization but not virus attachment Cell 1993 73: 309–319

    Article  CAS  PubMed  Google Scholar 

  25. Clesham FJ, Browne H, Efstathiou S, Weissberg PL . Enhancer stimulation unmasks latent gene transfer after adenovirus-mediated gene delivery into human vascular smooth muscle cells Circ Res 1996 79: 1188–1195

    Article  CAS  PubMed  Google Scholar 

  26. Baldwin HS, Mickanin C, Buck CA . Adenoviral mediated gene transfer during initial organogenesis in the mammalian embryo is promoter-dependent and tissue-specific Gene Therapy 1997 4: 1142–1149

    Article  CAS  PubMed  Google Scholar 

  27. Rome JJ et al. Adenoviral vector-mediated gene transfer into sheep arteries using a double-balloon catheter Hum Gene Ther 1994 5: 1249–1258

    Article  CAS  PubMed  Google Scholar 

  28. Schulick AH et al. Endothelium-specific in vivo gene transfer Circ Res 1995 77: 475–485

    Article  CAS  PubMed  Google Scholar 

  29. Yang Y, Li Q, Ertl HCJ, Wilson JM . Cellular and humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses J Virol 1995 69: 2004–2015

    CAS  PubMed  PubMed Central  Google Scholar 

  30. Walter J et al. Successful expression of human factor IX following repeat administration of an adenoviral vector in mice Proc Natl Acad Sci USA 1996 93: 3056–3061

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Antica M, Wu L, Shortman K, Scollay R . Intrathymic lymphoid precursor cells during fetal thymus development J Immunol 1993 151: 5887–5895

    CAS  PubMed  Google Scholar 

  32. Muneoka K, Wanek N, Bryant SV . Mouse embryos develop normally ex utero J Exp Zoo 1996 239: 289–293

    Article  Google Scholar 

  33. Graham FL, Prevec L . Methods for construction of adenovirus vectors Mol Biotechnol 1995 3: 207–220

    Article  CAS  PubMed  Google Scholar 

  34. Hogan B, Beddington R, Costantini F, Lacy E (eds) . Manipulating the Mouse Embryo; a Laboratory Manual, edn 2 Cold Spring Harbor Laboratory Press: Cold Spring Harbor 1994

    Google Scholar 

  35. Bergelson JM et al. Isolation of a common receptor for Coxsackie B viruses and adenoviruses 2 and 5 Science 1997 275: 1320–1323

    Article  CAS  PubMed  Google Scholar 

  36. Wilkinson DG, Green J . In situ hybridization and the three-dimensional reconstruction of serial sections. In: Copp AJ, Cockroft DL (eds) Postimplantation Mammalian Embryos. A Practical Approach Oxford University Press: New York 1990 pp 155–171

    Google Scholar 

Download references

Acknowledgements

We would like to acknowledge the support of the Institute for Human Gene Therapy at the University of Pennsylvania, and Leslie Underkoffler for the in situ hybridization studies. This work was funded in part by AHA Fellowship (9604602S) to SKS; NIH A135667 and HL54734 to JMB; NIH CA19144–21, CA10815–27, HL51533, HL47670 to CAB; HL56582, DCD 02027, AHA 95014210 and a grant from the ML Smith Lead Charitable Trust to HSB. JMB and HSB are Established Investigators of the American Heart Association.

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Authors and Affiliations

  1. Division of Cardiology, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    S K Schachtner

  2. Division of Cardiology, The Wistar Institute and Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    C A Buck

  3. Division of Immunologic and Infectious Diseases, Children’s Hospital of Philadelphia, Philadelphia, PA, USA

    J M Bergelson

  4. Division of Cardiology, Children’s Hospital of Philadelphia and The Wistar Institute, Philadelphia, PA, USA

    H S Baldwin

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  1. S K Schachtner
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  4. H S Baldwin
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Schachtner, S., Buck, C., Bergelson, J. et al. Temporally regulated expression patterns following in utero adenovirus-mediated gene transfer. Gene Ther 6, 1249–1257 (1999). https://doi.org/10.1038/sj.gt.3300939

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