Skip to main content
SpringerLink
Log in

Adeno-Associated Viral Vector-Delivered Hypoxia-Inducible Gene Expression in Ischemic Hearts

  • Protocol
Cardiac Gene Expression

Part of the book series: Methods in Molecular Biology ((MIMB,volume 366))

Abstract

This chapter describes a system using adeno-associated viral (AAV) vector to deliver hypoxia-inducible gene expression to ischemic hearts. The hypoxia induction of gene expression in this system is based on the accumulation of hypoxia-inducible factor-1 (HIF-1) in ischemic hearts and the use of hypoxia-response element (HRE) identified from the enhancers of genes, the expression of which can be induced by hypoxia. The methods of plasmid and AAV vector construction for hypoxia-inducible gene expression, viral vector production and purification, and viral titer determination are described. This chapter also illustrates the methods that can be used to test hypoxia-inducible gene expression in vitro and in vivo, including hypoxia treatment of cultured cells, generation of murine ischemic heart models, and analysis of gene expression.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Jones, S. P., Greer, J. J., Kakkar, A. K., et al. (2004) Endothelial nitric oxide synthase overexpression attenuates myocardial reperfusion injury. Am. J. Physiol. Heart Circ. Physiol. 286, H276–H282.

    Article  CAS  PubMed  Google Scholar 

  2. Jones, S. P., Greer, J. J., van Haperen, R., Duncker, D. J., de Crom, R., and Lefer, D. J. (2003) Endothelial nitric oxide synthase overexpression attenuates congestive heart failure in mice. Proc. Natl. Acad. Sci. USA 100, 4891–4896.

    Article  CAS  PubMed  Google Scholar 

  3. Woo, Y. J., Zhang, J. C., Vijayasarathy, C., et al. (1998) Recombinant adenovirus-mediated cardiac gene transfer of Superoxide dismutase and catalase attenuates postischemic contractile dysfunction. Circulation 98, II255–II260; discussion II260–II261.

    CAS  PubMed  Google Scholar 

  4. Mack, C. A., Patel, S. R., Schwarz, E. A., et al. (1998) Biologic bypass with the use of adenovirus-mediated gene transfer of the complementary deoxyribonucleic acid for vascular endothelial growth factor 121 improves myocardial perfusion and function in the ischemic porcine heart. J. Thorac. Cardiovasc. Surg. 115, 168–176; discussion 176–177.

    Article  CAS  PubMed  Google Scholar 

  5. Giordano, F. J., Ping, P., McKirnan, M. D., et al. (1996) Intracoronary gene transfer of fibroblast growth factor-5 increases blood flow and contractile function in an ischemic region of the heart [see comments]. Nat. Med. 2, 534–539.

    Article  CAS  PubMed  Google Scholar 

  6. Laham, R. J., Simons, M., and Sellke, F. (2001) Gene transfer for angiogenesis in coronary artery disease. Annu. Rev. Med. 52, 485–502.

    Article  CAS  PubMed  Google Scholar 

  7. Banai, S., Jaklitsch, M. T., Shou, M., et al. (1994) Angiogenic-induced enhancement of collateral blood flow to ischemic myocardium by vascular endothelial growth factor in dogs. Circulation 89, 2183–2189.

    CAS  PubMed  Google Scholar 

  8. Lee, R. J., Springer, M. L., Blanco-Bose, W. E., Shaw, R., Ursell, P. C., and Blau, H. M. (2000) VEGF gene delivery to myocardium: deleterious effects of unregulated expression. Circulation 102, 898–901.

    CAS  PubMed  Google Scholar 

  9. Springer, M. L., Chen, A. S., Kraft, P. E., Bednarski, M., and Blau, H. M. (1998) VEGF gene delivery to muscle: potential role for vasculogenesis in adults. Mol. Cell 2, 549–558.

    Article  CAS  PubMed  Google Scholar 

  10. Dennery, P. A., Sridhar, K. J., Lee, C. S., et al. (1997) Heme oxygenase-mediated resistance to oxygen toxicity in hamster fibroblasts. J. Biol. Chem. 272, 14937–14942.

    Article  CAS  PubMed  Google Scholar 

  11. Suttner, D. M. and Dennery, P. A. (1999) Reversal of HO-1 related cytoprotection with increased expression is due to reactive iron. FASEB J. 13, 1800–1809.

    CAS  PubMed  Google Scholar 

  12. Bohl, D., Salvetti, A., Moullier, P., and Heard, J. M. (1998) Control of erythropoietin delivery by doxycycline in mice after intramuscular injection of adeno-associated vector. Blood 92, 1512–1517.

    CAS  PubMed  Google Scholar 

  13. Hofmann, A., Nolan, G. P., and Blau, H. M. (1996) Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette [see comments]. Proc. Natl. Acad. Sci. USA 93, 5185–5190.

    Article  CAS  PubMed  Google Scholar 

  14. Bohl, D., Naffakh, N., and Heard, J. M. (1997) Long-term control of erythropoietin secretion by doxycycline in mice transplanted with engineered primary myoblasts [see comments]. Nat. Med. 3, 299–305.

    Article  CAS  PubMed  Google Scholar 

  15. Gossen, M. and Bujard, H. (1992) Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc. Natl. Acad. Sci. USA 89, 5547–5551.

    Article  CAS  PubMed  Google Scholar 

  16. Wang, Y., O’Malley, B. W., Jr., Tsai, S. Y., and O’Malley, B. W. (1994) A regulatory system for use in gene transfer. Proc. Natl. Acad. Sci. USA 91, 8180–8184.

    Article  CAS  PubMed  Google Scholar 

  17. Blau, H. M. and Rossi, F. M. (1999) Tet B or not tet B: advances in tetracycline-inducible gene expression [comment]. Proc. Natl. Acad. Sci. USA 96, 797–799.

    Article  CAS  PubMed  Google Scholar 

  18. Rossi, F. M., Guicherit, O. M., Spicher, A., et al. (1998) Tetracycline-regulatable factors with distinct dimerization domains allow reversible growth inhibition by pl6. Nat. Genet. 20, 389–393.

    Article  CAS  PubMed  Google Scholar 

  19. Kringstein, A. M., Rossi, F. M., Hofmann, A., and Blau, H. M. (1998) Graded transcriptional response to different concentrations of a single transactivator. Proc. Natl. Acad. Sci. USA 95, 13670–13675.

    Article  CAS  PubMed  Google Scholar 

  20. Wang, G. L., Jiang, B. H., Rue, E. A., and Semenza, G. L. (1995) Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc. Natl. Acad. Sci. USA 92, 5510–5514.

    Article  CAS  PubMed  Google Scholar 

  21. Wang, G. L. and Semenza, G. L. (1995) Purification and characterization of hypoxia-inducible factor 1. J. Biol. Chem. 270, 1230–1237.

    Article  CAS  PubMed  Google Scholar 

  22. Jiang, B. H., Rue, E., Wang, G. L., Roe, R., and Semenza, G. L. (1996) Dimerization, DNA binding, and transactivation properties of hypoxia-inducible factor 1. J. Biol. Chem. 271, 17771–17778.

    Article  CAS  PubMed  Google Scholar 

  23. Su, H., Arakawa-Hoyt, J., and Kan, Y. W. (2002) Adeno-associated viral vector-mediated hypoxia response element-regulated gene expression in mouse ischemic heart model. Proc. Natl. Acad. Sci. USA 99, 9480–9485.

    Article  CAS  PubMed  Google Scholar 

  24. Pachori, A. S., Melo, L. G., Hart, M. L., et al. (2004) Hypoxia-regulated therapeutic gene as a preemptive treatment strategy against ischemia/reperfusion tissue injury. Proc. Natl. Acad. Sci. USA 101, 12282–12287.

    Article  CAS  PubMed  Google Scholar 

  25. Su, H., Lu, R., and Kan, Y. W. (2000) Adeno-associated viral vector-mediated vascular endothelial growth factor gene transfer induces neovascular formation in ischemic heart. Proc. Natl. Acad. Sci. USA 97, 13801–13806.

    Article  CAS  PubMed  Google Scholar 

  26. Du, L., Kido, M., Lee, D. V., et al. (2004) Differential myocardial gene delivery by recombinant serotype-specific adeno-associated viral vectors. Mol Ther. 10, 604–608.

    Article  CAS  PubMed  Google Scholar 

  27. Semenza, G. L. and Wang, G. L. (1992) A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol. Cell Biol. 12, 5447–5454.

    CAS  PubMed  Google Scholar 

  28. Wang, G. L. and Semenza, G. L. (1993) General involvement of hypoxia-inducible factor 1 in transcriptional response to hypoxia. Proc. Natl. Acad. Sci. USA 90, 4304–4308.

    Article  CAS  PubMed  Google Scholar 

  29. Chavez, J. C., Agani, F., Pichiule, P., and LaManna, J. C. (2000) Expression of hypoxia-inducible factor-1 alpha in the brain of rats during chronic hypoxia. J. Appl Physiol. 89, 1937–1942.

    CAS  PubMed  Google Scholar 

  30. Ruan, H., Su, H., Hu, L., Lamborn, K. R., Kan, Y. W., and Deen, D. F. (2001) A hypoxia-regulated adeno-associated virus vector for cancer-specific gene therapy. Neoplasia 3, 255–263.

    Article  CAS  PubMed  Google Scholar 

  31. Shibata, T., Giaccia, A. J., and Brown, J. M. (2000) Development of a hypoxia-responsive vector for tumor-specific gene therapy. Gene Ther. 7, 493–498.

    Article  CAS  PubMed  Google Scholar 

  32. Gao, G. P., Alvira, M. R., Wang, L., Calcedo, R., Johnston, J., and Wilson, J. M. (2002) Novel adeno-associated viruses from rhesus monkeys as vectors for human gene therapy. Proc. Natl Acad. Sci. USA 99, 11854–11859.

    Article  CAS  PubMed  Google Scholar 

  33. Rutledge, E. A., Halbert, C. L., and Russell, D. W. (1998) Infectious clones and vectors derived from adeno-assoicated virus (AAV) serotypes other than AAV type 2. J. Virol 72, 309–319.

    CAS  PubMed  Google Scholar 

  34. Muramatsu, S., Mizukami, H., Young, N. S., and Brown, K. E. (1996) Nucleotide sequencing and generation of an infectious clone of adeno-associated virus 3. Virology 221, 208–217.

    Article  CAS  PubMed  Google Scholar 

  35. Chiorini, J. A., Yang, L., Liu, Y., Safer, B., and Kotin, R. M. (1997) Cloning of adeno-associated virus type 4 (AAV4) and generation of recombinant AAV4 particles. J. Virol 71, 6823–6833.

    CAS  PubMed  Google Scholar 

  36. Chiorini, J. A., Kim, F., Yang, L., and Kotin, R. M. (1999) Cloning and characterization of adeno-associated virus type 5. J. Virol 73, 1309–1319.

    CAS  PubMed  Google Scholar 

  37. Xiao, W., Chirmule, N., Berta, S. C., McCullough, B., Gao, G., and Wilson, J. M. (1999) Gene therapy vectors based on adeno-associated virus type 1. J. Virol 73, 3994–4003.

    CAS  PubMed  Google Scholar 

  38. Du, L., Sullivan, C. C., Chu, D., et al. (2003) Signaling molecules in nonfamilial pulmonary hypertension. N. Engl. J. Med. 348, 500–509.

    Article  CAS  PubMed  Google Scholar 

  39. Clark, K. R., Liu, X., McGrath, J. P., and Johnson, P. R. (1999) Highly purified recombinant adeno-associated virus vectors are biologically active and free of detectable helper and wild-type viruses. Hum. Gene Ther. 10, 1031–1039.

    Article  CAS  PubMed  Google Scholar 

  40. Zolotukhin, S., Potter, M., Zolotukhin, L, et al. (2002) Production and purification of serotype 1, 2, and 5 recombinant adeno-associated viral vectors. Methods 28, 158–167.

    Article  CAS  PubMed  Google Scholar 

  41. Zolotukhin, S., Byrne, B. J., Mason, E., et al. (1999) Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield. Gene Ther. 6, 973–985.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Cardiovascular Research Institute, University of California, San Francisco, San Francisco, CA, USA

    Hua Su & Yuet Wai Kan

  2. Department of Medicine, University of California, San Francisco, San Francisco, CA, USA

    Hua Su & Yuet Wai Kan

  3. Department of Laboratory Medicine, University of California, San Francisco, San Francisco, CA, USA

    Yuet Wai Kan

Authors
  1. Hua Su
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yuet Wai Kan
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Division of Cardiology, CVRL/Geffen School of Medicine at UCLA, Los Angeles, CA

    Jun Zhang

  2. Division of Cardiology, University of Louisville, Louisville, KY

    Gregg Rokosh

Rights and permissions

Reprints and permissions

Copyright information

© 2007 Humana Press Inc.

About this protocol

Cite this protocol

Su, H., Kan, Y.W. (2007). Adeno-Associated Viral Vector-Delivered Hypoxia-Inducible Gene Expression in Ischemic Hearts. In: Zhang, J., Rokosh, G. (eds) Cardiac Gene Expression. Methods in Molecular Biology, vol 366. Humana Press. https://doi.org/10.1007/978-1-59745-030-0_19

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-030-0_19

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-352-7

  • Online ISBN: 978-1-59745-030-0

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation