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Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice

Gene Therapy volume 8, pages 977–986 (2001)Cite this article

Abstract

We investigated genes expression by retrograde axonal transport of replication-defective adenoviruses carrying genes for LacZ (AdLacZ) and Bcl-2 in motor neurons of transgenic mice expressing mutant human Cu/Zn superoxide dismutase (SOD1) gene containing a substitution of alanine for glycine at position 93. We found that intramuscular injection of AdLacZ into the tongue of mutant SOD1 transgenic mice and their wild-type littermates at various ages results in high expression of the transgene and similar time course of expression in hypoglossal cranial nerve nuclei, suggesting no difference in the behavior of the transgene expression between the two groups. Subsequently, we employed a molecular switching cassette for Bcl-2 designed to express Bcl-2 by Cre-loxP recombination using adenoviral vectors, and examined the COS7 and primary neuronal cells with the mutant SOD1 gene. The overexpression of Bcl-2 in both cells and the neuronal protection against staurosporine-induced apoptosis were observed, after dual infection of adenoviral vectors with cassette for Bcl-2 (AxCALNLBcl-2) and Cre recombinase (AxCANCre). After inoculation of AxCALNLBcl-2 followed by AxCANCre into the tongue of both mutant SOD1 transgenic mice and wild-type littermates, Bcl-2 was detected in both the injection site and the hypoglossal nuclei of brainstems, suggesting that this was the result of retrograde transport of AxCALNLBcl-2 and AxCANCre and expression of Bcl-2 by Cre recombinase in the hypoglossal nuclei. This strategy for delivery of exogenous genes such as Bcl-2 will be useful for studying neuronal death/survival and introducing foreign genes into postmitotic motor neurons, and in gene therapy for motor neuron diseases such as ALS.

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References

  1. Williams DB, Windebank AJ . Motor neuron disease (amyotrophic lateral sclerosis) Mayo Clin Proc 1991 66: 54–82

    Article  CAS  PubMed  Google Scholar 

  2. Rosen DR et al. Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis Nature 1993 362: 59–62

    Article  CAS  PubMed  Google Scholar 

  3. Mulder DW, Kurland LT, Offord KP, Beard CM . Familial adult motor neuron disease: amyotrophic lateral sclerosis Neurology 1986 36: 511–517

    Article  CAS  PubMed  Google Scholar 

  4. Gurney ME et al. Motor neuron degeneration in mice that express a human Cu,Zn superoxide dismutase mutation Science 1994 264: 1772–1775

    Article  CAS  PubMed  Google Scholar 

  5. Ripps ME et al. Transgenic mice expressing an altered murine superoxide dismutase gene provide an animal model of amyotrophic lateral sclerosis Proc Natl Acad Sci USA 1995 92: 689–693

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Dal Canto MC, Gurney ME . Development of central nervous system pathology in a murine transgenic model of human amyotrophic lateral sclerosis Am J Pathol 1994 145: 1271–1279

    CAS  PubMed  PubMed Central  Google Scholar 

  7. Rothstein JD . Therapeutic horizons for amyotrophic lateral sclerosis Curr Opin Neurobiol 1996 6: 679–687

    Article  CAS  PubMed  Google Scholar 

  8. Aebischer P et al. Intrathecal delivery of CNTF using encapsulated genetically modified xenogeneic cells in amyotrophic lateral sclerosis patients Nat Med 1996 2: 696–699

    Article  CAS  PubMed  Google Scholar 

  9. Haase G et al. Gene therapy of murine motor neuron disease using adenoviral vectors for neurotrophic factors Nat Med 1997 3: 429–436

    Article  CAS  PubMed  Google Scholar 

  10. Azzouz M et al. Increased motoneuron survival and improved neuromuscular function in transgenic ALS mice after intraspinal injection of an adeno-associated virus encoding Bcl-2 Hum Mol Genet 2000 9: 803–811

    Article  CAS  PubMed  Google Scholar 

  11. Costantini LC, Bakowska JC, Breakefield XO, Isacson O . Gene therapy in the CNS Gene Therapy 2000 7: 93–109

    Article  CAS  PubMed  Google Scholar 

  12. Akli S et al. Transfer of a foreign gene into the brain using adenovirus vectors Nat Genet 1993 3: 224–228

    Article  CAS  PubMed  Google Scholar 

  13. Davidson BL et al. A model system for in vivo gene transfer into the central nervous system using an adenoviral vector Nat Genet 1993 3: 219–223

    Article  CAS  PubMed  Google Scholar 

  14. Le Gal La Salle G et al. An adenovirus vector for gene transfer into neurons and glia in the brain Science 1993 259: 988–990

    Article  CAS  PubMed  Google Scholar 

  15. Byrnes AP, Rusby JE, Wood MJ, Charlton HM . Adenovirus gene transfer causes inflammation in the brain Neuroscience 1995 66: 1015–1024

    Article  CAS  PubMed  Google Scholar 

  16. Ohmoto Y et al. Variation in the immune response to adenoviral vectors in the brain: influence of mouse strain, environmental conditions and priming Gene Therapy 1999 6: 471–481

    Article  CAS  PubMed  Google Scholar 

  17. Ghadge GD et al. CNS gene delivery by retrograde transport of recombinant replication-defective adenoviruses Gene Therapy 1995 2: 132–137

    CAS  PubMed  Google Scholar 

  18. Ho DY, McLaughlin JR, Sapolsky RM . Inducible gene expression from defective herpes simplex virus vectors using the tetracycline-responsive promoter system Mol Brain Res 1996 41: 200–209

    Article  CAS  PubMed  Google Scholar 

  19. Oligino T et al. Drug inducible transgene expression in brain using a herpes simplex virus vector Gene Therapy 1998 5: 491–496

    Article  CAS  PubMed  Google Scholar 

  20. Sato N et al. A novel strategy for introducing exogenous bcl-2 into neuronal cells: the Cre/loxP system-mediated activation of bcl-2 for preventing programmed cell death using recombinant adenoviruses Mol Cell Neurosci 1998 12: 65–78

    Article  CAS  PubMed  Google Scholar 

  21. Warita H, Abe K, Setoguchi Y, Itoyama Y . Expression of adenovirus-mediated E. coli lacZ gene in skeletal muscles and spinal motor neurons of transgenic mice with a mutant superoxide dismutase gene Neurosci Lett 1998 246: 153–156

    Article  CAS  PubMed  Google Scholar 

  22. Warita H, Itoyama Y, Abe K . Selective impairment of fast anterograde axonal transport in the peripheral nerves of asymptomatic transgenic mice with a G93A mutant SOD1 gene Brain Res 1999 819: 120–131

    Article  CAS  PubMed  Google Scholar 

  23. Anderson WF . Human gene therapy Nature 1998 392 (Suppl.): 25–30

    Google Scholar 

  24. Navarro V et al. Efficient gene transfer and long-term expression in neurons using a recombinant adenovirus with a neuron-specific promoter Gene Therapy 1999 6: 1884–1892

    Article  CAS  PubMed  Google Scholar 

  25. Niwa H, Yamamura K, Miyazaki J . Efficient selection for high-expression transfectants with a novel eukaryotic vector Gene 1991 108: 193–199

    Article  CAS  PubMed  Google Scholar 

  26. Rabizadeh S et al. Mutations associated with amyotrophic lateral sclerosis convert superoxide dismutase from an antiapoptotic gene to a proapoptotic gene: studies in yeast and neural cells Proc Natl Acad Sci USA 1995 92: 3024–3028

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Kostic V et al. Bcl-2: prolonging life in a transgenic mouse model of familial amyotrophic lateral sclerosis Science 1997 277: 559–562

    Article  CAS  PubMed  Google Scholar 

  28. Kanegae Y et al. Efficient gene activation system on mammalian cell chromosomes using recombinant adenovirus producing Cre recombinase Gene 1996 181: 207–212

    Article  CAS  PubMed  Google Scholar 

  29. Kanegae Y et al. Efficient gene activation in mammalian cells by using recombinant adenovirus expressing site-specific Cre recombinase Nucleic Acids Res 1995 23: 3816–3821

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Korsmeyer SJ . Bcl-2 initiates a new category of oncogenes: regulators of cell death Blood 1992 80: 879–886

    CAS  PubMed  Google Scholar 

  31. Ghadge GD et al. Mutant superoxide dismutase-1-linked familial amyotrophic lateral sclerosis: molecular mechanisms of neuronal death and protection J Neurosci 1997 17: 8756–8766

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Pasinelli P et al. Caspase-1 is activated in neural cells and tissue with amyotrophic lateral sclerosis-associated mutations in copper-zinc superoxide dismutase Proc Natl Acad Sci USA 1998 95: 15763–15768

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Mena MA et al. Effects of wild-type and mutated copper/zinc superoxide dismutase on neuronal survival and L-DOPA-induced toxicity in postnatal midbrain culture J Neurochem 1997 69: 21–33

    Article  CAS  PubMed  Google Scholar 

  34. Finiels F et al. Specific and efficient gene transfer strategy offers new potentialities for the treatment of motor neurone diseases Neuroreport 1995 7: 373–378

    Article  CAS  PubMed  Google Scholar 

  35. Namikawa K et al. Akt/protein kinase B prevents injury-induced motoneuron death and accelerates axonal regeneration J Neurosci 2000 20: 2875–2886

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Tamatani M et al. Tumor necrosis factor induces Bcl-2 and Bcl-x expression through NFkappaB activation in primary hippocampal neurons J Biol Chem 1999 274: 8531–8538

    Article  CAS  PubMed  Google Scholar 

  37. Morsy MA et al. Efficient adenoviral-mediated ornithine transcarbamylase expression in deficient mouse and human hepatocytes J Clin Invest 1993 92: 1580–1586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Mita S et al. Detection of ‘deleted’ mitochondrial genomes in cytochrome-c oxidase-deficient muscle fibers of a patient with Kearns–Sayre syndrome Proc Natl Acad Sci USA 1989 86: 9509–9513

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Mita S et al. Single muscle fiber analysis of myoclonus epilepsy with ragged-red fibers Muscle Nerve 1998 21: 490–497

    Article  CAS  PubMed  Google Scholar 

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Author notes

  1. S Yamashita and S Mita: The first two authors contributed equally to this work

Authors and Affiliations

  1. Department of Neurology, Kumamoto University, School of Medicine, Kumamoto, Japan

    S Yamashita, S Mita, T Arima, Y Maeda, E Kimura, Y Nishida, T Murakami & M Uchino

  2. Department of Neurobiology, Tokyo Metropolitan Institute for Neuroscience, Tokyo, Japan

    H Okado

  3. Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology, Saitama, Kawaguchi, Japan

    H Okado

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  1. S Yamashita
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Yamashita, S., Mita, S., Arima, T. et al. Bcl-2 expression by retrograde transport of adenoviral vectors with Cre-loxP recombination system in motor neurons of mutant SOD1 transgenic mice. Gene Ther 8, 977–986 (2001). https://doi.org/10.1038/sj.gt.3301479

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