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
Link, C. D. (2001) Transgenic invertebrate models of age-associated neurodegenerative diseases. Mech. Ageing Dev. 122, 1639–1649.
Sipione, S. and Cattaneo, E. (2001) Modeling huntington’s disease in cells, flies, and mice. Mol. Neurobiol. 23, 21–51.
Zoghbi, H. and Botas, J. (2002) Mouse and fly models of neurodegeneration. Trends Genet. 18, 463.
Cattaneo, E. and Conti, L. (1998) Generation and characterization of embryonic striatal conditionally immortalized ST14A cells. J. Neurosci. Res. 53, 223–234.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Hsich, G., Sena-Esteves, M., and Breakefield, X. O. (2002) Critical issues in gene therapy for neurologic disease. Hum. Gene Ther. 13, 579–604.
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.
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.
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.
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.
Kafri, T., van Praag, H., Gage, F. H., et al. (2000) Lentiviral vectors: regulated gene expression. Mol. Ther. 1, 516–521.
Mansuy, I. M. F.-B. and Bujard, H. (2000) Tetracycline-regulated gene expression in the brain. Curr. Opin. Neurobiol. 10, 593–596.
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.
Washbourne, P. and McAllister, A. K. (2002) Techniques for gene transfer into neurons. Curr. Opin. Neurobiol. 12, 566–573.
Zufferey, R., Nagy, D., Mandel, R. J., et al. (1997) Multiply attenuated lentiviral vector achieves efficient gene delivery in vivo. Nature 15, 871–875.
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.
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.
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.
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.
Negre, D., Duisit, G., Mangeot, P. E., et al. (2002) Lentiviral vectors derived from simian immunodeficiency virus. Curr. Topics Microbiol. Immunol. 261, 53–74.
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.
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.
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.
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.
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.
Dull, T., Zufferey, R., Kelly, M., et al. (1998) A third-generation lentivirus vector with a conditional packaging system. J. Virol. 72, 8463–8471.
Deglon, N. and Aebischer, P. (2002) Lentiviruses as vectors for CNS diseases. Curr. Topics Microbiol. Immunol. 261, 191–209.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Scherr, M., Battmer, K., Blomer, U., et al. (2001) Quantitative determination of lentiviral vector particle numbers by real-time PCR. Biotechniques 31, 520–524.
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.
Messier, C., Emond, S., and Ethier, K. (1999) New techniques in stereotaxic surgery and anesthesia in the mouse. Pharmacol. Biochem. Behav. 63, 313–318.
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.
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.
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.
Reiser, J. (2000) Production and concentration of pseudotyped HIV-1-based gene transfer vectors. Gene Ther. 7, 910–913.
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.
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.
Connolly, J. B. (2002) Lentiviruses in gene therapy clinical research. Gene Ther. 9, 1730–1734.
Follenzi, A. and Naldini, L. (2002) HIV-based vectors. Preparation and use. Methods Mol. Med. 69, 259–274.
Higashikawa, F. and Chang, L. (2001) Kinetic analyses of stability of simple and complex retroviral vectors. Virology 280, 124–131.
Clavel, F. and Charneau, P. (1994) Fusion from without directed by human immunodeficiency virus particles. J. Virol. 68, 1179–1185.
Delenda, C., Audit, M., and Danos, O. (2002) Biosafety issues in lentivector production. Curr. Topics Microbiol. Immunol. 261, 123–141.
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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
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