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Trinucleotide Repeat Protocols pp 199–213Cite as

Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disorders

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 277))

Summary

This chapter describes the potential use of viral-mediated gene transfer in the central nervous system as a new strategy in developing animal models of neurodegenerative diseases. To illustrate the approach, procedures for the production of lentiviral vectors encoding polyQ proteins are provided, as well as methods for the determination of viral titers, in vitro infection, and basic protocols for in vivo studies in rodents.

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References

  1. Link, C. D. (2001) Transgenic invertebrate models of age-associated neurodegenerative diseases. Mech. Ageing Dev. 122, 1639–1649.

    Article  PubMed  CAS  Google Scholar 

  2. Sipione, S. and Cattaneo, E. (2001) Modeling huntington’s disease in cells, flies, and mice. Mol. Neurobiol. 23, 21–51.

    Article  PubMed  CAS  Google Scholar 

  3. Zoghbi, H. and Botas, J. (2002) Mouse and fly models of neurodegeneration. Trends Genet. 18, 463.

    Article  PubMed  CAS  Google Scholar 

  4. Cattaneo, E. and Conti, L. (1998) Generation and characterization of embryonic striatal conditionally immortalized ST14A cells. J. Neurosci. Res. 53, 223–234.

    Article  PubMed  CAS  Google Scholar 

  5. Saudou, F., Finkbeiner, S., Devys, D., et al. (1998) Huntingtin acts in the nucleus to induce apoptosis but death does not correlate with the formation of intranuclear inclusions. Cell 95, 55–66.

    Article  PubMed  CAS  Google Scholar 

  6. Lunkes, A. and Mandel, J. L. (1998) A cellular model that recapitulates major pathogenic steps of Huntington’s disease. Hum. Mol. Genet. 7, 1355–1361.

    Article  PubMed  CAS  Google Scholar 

  7. 7 Ho, L. W., Brown, R., Maxwell, M., et al. (2001) Wild type Huntingtin reduces the cellular toxicity of mutant Huntingtin in mammalian cell models of Huntington’s disease. J. Med. Genet. 38, 450–452.

    Article  PubMed  CAS  Google Scholar 

  8. Senut, M. C., Suhr, S. T., Kaspar, B., et al. (2000) Intraneuronal aggregate formation and cell death after viral expression of expanded polyglutamine tracts in the adult rat brain. J. Neurosci. 20, 219–229.

    PubMed  CAS  Google Scholar 

  9. de Almeida, L. P., Ross, C. A., Zala, D., et al. (2002) Lentiviral-mediated delivery of mutant huntingtin in the striatum of rats induces a selective neuropathology modulated by polyglutamine repeat size, huntingtin expression levels, and protein length. J. Neurosci. 22, 3473–3483.

    PubMed  Google Scholar 

  10. Klein, R. L., King, M. A., Hamby, M. E., et al. (2002) Dopaminergic cell loss induced by human A30P alpha-synuclein gene transfer to the rat substantia nigra. Hum. Gene Ther. 13, 605–612.

    Article  PubMed  CAS  Google Scholar 

  11. Lo Bianco, C., Ridet, J. L., Schneider, B. L., et al. (2002) Alpha-synucleinopathy and selective dopaminergic neuron loss in a rat lentiviral-based model of Parkinson’s disease. Proc. Natl. Acad. Sci. USA 99, 10,813–10,818.

    Article  PubMed  Google Scholar 

  12. Kirik, D., Rosenblad, C., Burger, C., et al. (2002) Parkinson-like neurodegeneration induced by targeted overexpression of alpha-synuclein in the nigrostriatal system. J. Neurosci. 22, 2780–2791.

    PubMed  CAS  Google Scholar 

  13. Kirik, D., Annett, L. E., Burger, C., et al. (2003) Nigrostriatal alpha-synucleinopathy induced by viral vector-mediated overexpression of human alpha-synuclein: a new primate model of Parkinson’s disease. Proc. Natl. Acad. Sci. USA 100, 2884–2889.

    Article  PubMed  CAS  Google Scholar 

  14. Janson, C. G., McPhee, S. W., Leone, P., et al. (2001) Viral-based gene transfer to the mammalian CNS for functional genomic studies. Trends Neurosci. 24, 706–712.

    Article  PubMed  CAS  Google Scholar 

  15. Hsich, G., Sena-Esteves, M., and Breakefield, X. O. (2002) Critical issues in gene therapy for neurologic disease. Hum. Gene Ther. 13, 579–604.

    Article  PubMed  CAS  Google Scholar 

  16. Kay, M. A., Glorioso, J. C., and Naldini, L. (2001) Viral vectors for gene therapy: the art of turning infectious agents into vehicles of therapeutics. Nature Med. 7, 33–40.

    Article  PubMed  CAS  Google Scholar 

  17. Hurlbert, M. S., Zhou, W., Wasmeier, C., et al. (1999) Mice transgenic for an expanded CAG repeat in the Huntington’s disease gene develop diabetes. Diabetes 48, 649–651.

    Article  PubMed  CAS  Google Scholar 

  18. Mangiarini, L. F.-S., Sathasivam, K. F.-S., Seller, M. F.-C., et al. (1996) Exon 1 of the HD gene with an expanded CAG repeat is sufficient to cause a progressive neurological phenotype in transgenic mice. Cell 87(3), 493–506.

    Article  PubMed  CAS  Google Scholar 

  19. Reddy, P. H., Williams, M., and Tagle, D. A. (1999) Recent advances in understanding the pathogenesis of Huntington’s disease. Trends Neurosci. 22, 248–255.

    Article  PubMed  CAS  Google Scholar 

  20. Kafri, T., van Praag, H., Gage, F. H., et al. (2000) Lentiviral vectors: regulated gene expression. Mol. Ther. 1, 516–521.

    Article  PubMed  CAS  Google Scholar 

  21. Mansuy, I. M. F.-B. and Bujard, H. (2000) Tetracycline-regulated gene expression in the brain. Curr. Opin. Neurobiol. 10, 593–596.

    Article  PubMed  CAS  Google Scholar 

  22. Regulier, E., Pereira de Almeida, L., Sommer, B., et al. (2002) Dose-dependent neuroprotective effect of ciliary neurotrophic factor delivered via tetracycline-regulated lentiviral vectors in the quinolinic acid rat model of Huntington’s disease. Hum. Gene Ther. 13, 1981–1990.

    Article  PubMed  CAS  Google Scholar 

  23. Washbourne, P. and McAllister, A. K. (2002) Techniques for gene transfer into neurons. Curr. Opin. Neurobiol. 12, 566–573.

    Article  PubMed  CAS  Google Scholar 

  24. Zufferey, R., Nagy, D., Mandel, R. J., et al. (1997) Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nature 15, 871–875.

    Article  CAS  Google Scholar 

  25. Naldini, L., Blomer, U., Gallay, P., et al. (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272, 263–267.

    Article  PubMed  CAS  Google Scholar 

  26. Naldini, L., Blomer, U., Gage, F. H., et al. (1996) Efficient transfer, integration, and sustained long-term expression of the transgene in adult rat brains injected with a lentiviral vector. Proc. Natl. Acad. Sci. USA 93, 11,382–11,388.

    Article  PubMed  CAS  Google Scholar 

  27. Poeschla, E. M., Wong-Staal, F., and Looney, D. J. (1998) Efficient transduction of nondividing human cells by feline immunodeficiency virus lentiviral vectors. Nature Med. 4, 354–357.

    Article  PubMed  CAS  Google Scholar 

  28. Rohll, J. B., Mitrophanous, K. A., Martin-Rendon, E., et al. (2002) Design, production, safety, evaluation, and clinical applications of nonprimate lentiviral vectors. Methods Enzymol. 346, 466–500.

    Article  PubMed  CAS  Google Scholar 

  29. Negre, D., Duisit, G., Mangeot, P. E., et al. (2002) Lentiviral vectors derived from simian immunodeficiency virus. Curr. Topics Microbiol. Immunol. 261, 53–74.

    CAS  Google Scholar 

  30. Zufferey, R., Donello, J. E., Trono, D., et al. (1999) Woodchuck hepatitis virus posttranscriptional regulatory element enhances expression of transgenes delivered by retroviral vectors. J. Virol. 73, 2886–2892.

    PubMed  CAS  Google Scholar 

  31. Zennou, V., Petit, C., Guetard, D., et al. (2000) HIV-1 genome nuclear import is mediated by a central DNA flap. Cell 101, 173–185.

    Article  PubMed  CAS  Google Scholar 

  32. Follenzi, A., Ailles, L. E., Bakovic, S., et al. (2000) Gene transfer by lentiviral vectors is limited by nuclear translocation and rescued by HIV-1 pol sequences. Nature Genet. 25, 217–222.

    Article  PubMed  CAS  Google Scholar 

  33. Sirven, A., Pflumio, F., Zennou, V., et al. (2000) The human immunodeficiency virus type-1 central DNA flap is a crucial determinant for lentiviral vector nuclear import and gene transduction of human hematopoietic stem cells. Blood 96, 4103–4110.

    PubMed  CAS  Google Scholar 

  34. Deglon, N., Tseng, J. L., Bensadoun, J. C., et al. (2000) Self-inactivating lentiviral vectors with enhanced transgene expression as potential gene transfer system in Parkinson’s disease. Hum. Gene Ther. 11, 179–190.

    Article  PubMed  CAS  Google Scholar 

  35. Dull, T., Zufferey, R., Kelly, M., et al. (1998) A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471.

    PubMed  CAS  Google Scholar 

  36. Deglon, N. and Aebischer, P. (2002) Lentiviruses as vectors for CNS diseases. Curr. Topics Microbiol. Immunol. 261, 191–209.

    CAS  Google Scholar 

  37. Blomer, U., Kafri, T., Randolph-Moore, L., et al. (1998) Bcl-xL protects adult septal cholinergic neurons from axotomized cell death. Proc. Natl. Acad. Sci. USA 95, 2603–2608.

    Article  PubMed  CAS  Google Scholar 

  38. Farson, D., Witt, R., McGuinness, R., et al. (2001) A new-generation stable inducible packaging cell line for lentiviral vectors. Hum. Gene Ther. 12, 981–997.

    Article  PubMed  CAS  Google Scholar 

  39. Yang, S., Delgado, R., King, S. R., et al. (1999) Generation of retroviral vector for clinical studies using transient transfection. Hum. Gene Ther. 10, 123–132.

    Article  PubMed  CAS  Google Scholar 

  40. Mitrophanous, K., Yoon, S., Rohll, J., et al. (1999) Stable gene transfer to the nervous system using a non-primate lentiviral vector. Gene Ther. 6, 1808–1818.

    Article  PubMed  CAS  Google Scholar 

  41. Negre, D., Mangeot, P. E., Duisit, G., et al. (2000) Characterization of novel safe lentiviral vectors derived from simian immunodeficiency virus (SIVmac251) that efficiently transduce mature human dendritic cells. Gene Ther. 7, 1613–1623.

    Article  PubMed  CAS  Google Scholar 

  42. Stitz, J., Buchholz, C. J., Engelstadter, M., et al. (2000) Lentiviral vectors pseudotyped with envelope glycoproteins derived from gibbon ape leukemia virus and murine leukemia virus 10A1. Virology 273, 16–20.

    Article  PubMed  CAS  Google Scholar 

  43. Duisit, G., Conrath, H., Saleun, S., et al. (2002) Five recombinant simian immunodeficiency virus pseudotypes lead to exclusive transduction of retinal pigmented epithelium in rat. Mol. Ther. 6, 446–454.

    Article  PubMed  CAS  Google Scholar 

  44. Kumar, M., Bradow, B. P., and Zimmerberg, J. (2003) Large-scale production of pseudotyped lentiviral vectors using baculovirus GP64. Hum. Gene Ther. 14, 67–77.

    Article  PubMed  CAS  Google Scholar 

  45. Sastry, L., Johnson, T., Hobson, M. J., et al. (2002) Titering lentiviral vectors: comparison of DNA, RNA and marker expression methods. Gene Ther. 9, 1155–1162.

    Article  PubMed  CAS  Google Scholar 

  46. Scherr, M., Battmer, K., Blomer, U., et al. (2001) Quantitative determination of lentiviral vector particle numbers by real-time PCR. Biotechniques 31, 520–524.

    PubMed  CAS  Google Scholar 

  47. Bensadoun, J. C., Mirochnitchenko, O., Inouye, M., et al. (1998) Attenuation of 6-OHDAinduced neurotoxicity in glutathione peroxidase transgenic mice. Eur. J. Neurosci. 10, 3231–3236.

    Article  PubMed  CAS  Google Scholar 

  48. Messier, C., Emond, S., and Ethier, K. (1999) New techniques in stereotaxic surgery and anesthesia in the mouse. Pharmacol. Biochem. Behav. 63, 313–318.

    Article  PubMed  CAS  Google Scholar 

  49. Paxinos, G., Watson, C., Pennisi, M., et al. (1985) Bregma, lambda and the interaural midpoint in stereotaxic surgery with rats of different sex, strain and weight. J. Neurosci. Methods 13, 139–143.

    Article  PubMed  CAS  Google Scholar 

  50. Bensadoun, J. C., Deglon, N., Tseng, J. L., et al. (2000) Lentiviral vectors as a gene delivery system in the mouse midbrain: cellular and behavioral improvements in a 6-OHDA model of Parkinson’s disease using GDNF. Exp. Neurol. 164(1), 15–24.

    Article  PubMed  CAS  Google Scholar 

  51. Richardson, J. H., Hofmann, W., Sodroski, J. G., et al. (1998) Intrabody-mediated knockout of the high-affinity IL-2 receptor in primary human T cells using a bicistronic lentivirus vector. Gene Ther. 5, 635–644.

    Article  PubMed  CAS  Google Scholar 

  52. Reiser, J. (2000) Production and concentration of pseudotyped HIV-1-based gene transfer vectors. Gene Ther. 7, 910–913.

    Article  PubMed  CAS  Google Scholar 

  53. Zhang, B., Xia, H. Q., Cleghorn, G., et al. (2001) A highly efficient and consistent method for harvesting large volumes of high-titre lentiviral vectors. Gene Ther. 8, 1745–1751.

    Article  PubMed  CAS  Google Scholar 

  54. Scherr, M., Battmer, K., Eder, M., et al. (2002) Efficient gene transfer into the CNS by lentiviral vectors purified by anion exchange chromatography. Gene Ther. 9, 1708–1714.

    Article  PubMed  CAS  Google Scholar 

  55. Connolly, J. B. (2002) Lentiviruses in gene therapy clinical research. Gene Ther. 9, 1730–1734.

    Article  PubMed  CAS  Google Scholar 

  56. Follenzi, A. and Naldini, L. (2002) HIV-based vectors. Preparation and use. Methods Mol. Med. 69, 259–274.

    PubMed  CAS  Google Scholar 

  57. Higashikawa, F. and Chang, L. (2001) Kinetic analyses of stability of simple and complex retroviral vectors. Virology 280, 124–131.

    Article  PubMed  CAS  Google Scholar 

  58. Clavel, F. and Charneau, P. (1994) Fusion from without directed by human immunodeficiency virus particles. J. Virol. 68, 1179–1185.

    PubMed  CAS  Google Scholar 

  59. Delenda, C., Audit, M., and Danos, O. (2002) Biosafety issues in lentivector production. Curr. Topics Microbiol. Immunol. 261, 123–141.

    CAS  Google Scholar 

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Author information

Authors and Affiliations

  1. Institute of Neuroscience, School of Life Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), Lausanne, Switzerland

    Etienne Régulier, Diana Zala, Patrick Aebischer & Nicole Déglon

Authors
  1. Etienne Régulier
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  2. Diana Zala
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  3. Patrick Aebischer
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  4. Nicole Déglon
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Editor information

Editors and Affiliations

  1. Life Science Division, Lawrence Berkeley Laboratory, Berkeley, CA

    Yoshinori Kohwi

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© 2004 Humana Press Inc.

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Régulier, E., Zala, D., Aebischer, P., Déglon, N. (2004). Lentiviral-Mediated Gene Transfer to Model Triplet Repeat Disorders. In: Kohwi, Y. (eds) Trinucleotide Repeat Protocols. Methods in Molecular Biology™, vol 277. Humana Press. https://doi.org/10.1385/1-59259-804-8:199

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  • DOI: https://doi.org/10.1385/1-59259-804-8:199

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-243-8

  • Online ISBN: 978-1-59259-804-5

  • eBook Packages: Springer Protocols

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