Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research Article
  • Published:

Modulation of neuronal survival and axonal growth in vivo by tetracycline-regulated neurotrophin expression

Gene Therapy volume 8, pages 954–960 (2001)Cite this article

Abstract

Vector systems for the regulated and reversible expression of therapeutic genes are likely to improve the safety and efficacy of gene therapy for medical disease. In the present study, we investigated whether the expression of genes transferred into the central nervous system by ex vivo gene therapy can be regulated in vivo leading to controlled neuronal survival and axonal growth. Primary rat fibroblasts were transfected with a retrovirus containing a tetracycline responsive promoter for the expression of the neurotrophin nerve growth factor (NGF) or green fluorescent protein as a control (GFP). After lesions of basal forebrain cholinergic neurons, NGF-mediated neuronal rescue and axonal growth could be completely controlled over a 2-week period by the addition or removal of the tetracycline modulator doxycycline in the animals’ drinking water. Further, continued expression of the reporter gene GFP could be reliably and repeatedly turned on and off in the injured CNS for at least 3 months post-grafting, the longest time point investigated. These data constitute the first report of regulated neuronal rescue and axonal growth by controlled neurotrophin gene delivery and long-term, regulated expression using ex vivo CNS gene therapy.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5

Similar content being viewed by others

References

  1. Tuszynski MH, Senut MC, Ray J, Roberts J . Somatic gene transfer to the adult primate central nervous system: in vitro and in vivo characterization of cells genetically modified to secrete nerve growth factor Neurobiol Dis 1994 1: 67–78

    Article  CAS  PubMed  Google Scholar 

  2. Kordower JH et al. Neurodegeneration prevented by lentiviral vector delivery of GDNF in primate models of Parkinson's disease Science 2000 290: 767–773

    Article  CAS  PubMed  Google Scholar 

  3. Simon DE, Roberts J, Gage FH, Tuszynski MH . Age-associated neuronal atrophy occurs in the primate brain and is reversible by growth factor gene therapy Proc Natl Acad Sci USA 1999 96: 10893–10898

    Article  Google Scholar 

  4. Blömer U et al. Applications of gene therapy to the CNS Hum Mol Genet 1996 5: 1397–1404

    Article  PubMed  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  6. Choi-Lundberg DL et al. Dopaminergic neurons protected from degeneration by GDNF gene therapy Science 1997 275: 838–841

    Article  CAS  PubMed  Google Scholar 

  7. Martinez-Serrano A et al. Long-term functional recovery from age-induced spatial memory impairments by nerve growth factor gene transfer to the rat basal forebrain Proc Natl Acad Sci USA 1996 93: 6355–6360

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Breakefield X, Jacobs A, Wang S . Genetic engineering for CNS regeneration. In: Tuszcynski MH, Kordower JH (eds) CNS Regeneration: Basic Science and Clinical Applications Academic Press: San Diego 1999 251–291

    Chapter  Google Scholar 

  9. Conner JM, Darracq MA, Roberts J, Tuszynski MH . Non-tropic actions of neurotrophins: subcortical NGF gene delivery reversess age-related degeneration of primate cortical cholinergic innervation Proc Natl Acad Sci USA 2001 98: 1941–1946

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Mohajeri MH, Figlewicz DA, Bohn MC . Intramuscular grafts of myoblasts genetically modified to secrete glial cell line-derived neurotrophic factor prevent motoneuron loss and disease progression in a mouse model of familial amyotrophic lateral sclerosis Hum Gene Ther 1999 10: 1853–1866

    Article  CAS  PubMed  Google Scholar 

  11. Gossen M et al. Transcriptional activation by tetracyclines in mammalian cells Science 1995 268: 1766–1769

    Article  CAS  PubMed  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Paulus W et al. Self-contained, tetracycline-regulated retroviral vector system for gene delivery to mammalian cells J Virol 1996 70: 62–67

    CAS  PubMed  PubMed Central  Google Scholar 

  14. Hofmann A, Nolan GP, Blau HM . Rapid retroviral delivery of tetracycline-inducible genes in a single autoregulatory cassette Proc Natl Acad Sci USA 1996 93: 5185–5190

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Hoshimaru M, Ray J, Sah DW, Gage FH . Differentiation of the immortalized adult neuronal progenitor cell line HC2S2 into neurons by regulatable suppression of the v-myc oncogene Proc Natl Acad Sci USA 1996 93: 1518–1523

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Blesch A, Uy H, Diergardt N, Tuszynski MH . Neurite outgrowth can be modulated in vitro using a tetracycline-repressible gene therapy vector expressing human nerve growth factor J Neurosci Res 2000 59: 402–409

    Article  CAS  PubMed  Google Scholar 

  17. Rosenberg MB et al. Grafting genetically modified cells to the damaged brain: restorative effects of NGF expression Science 1988 242: 1575–1578

    Article  CAS  PubMed  Google Scholar 

  18. Hefti F . Nerve growth factor promotes survival of septal cholinergic neurons after fimbrial transections J Neurosci 1986 6: 2155–2162

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Fischer W et al. Amelioration of cholinergic neuron atrophy and spatial memory impairment in aged rats by nerve growth factor Nature 1987 329: 65–68

    Article  CAS  PubMed  Google Scholar 

  20. Tuszynski MH, Armstrong DM, Gage FH . Basal forebrain cell loss following fimbria/fornix transection Brain Res 1990 508: 241–248

    Article  CAS  PubMed  Google Scholar 

  21. Kordower JH et al. The aged monkey basal forebrain: rescue and sprouting of axotomized basal forebrain neurons after grafts of encapsulated cells secreting human nerve growth factor Proc Natl Acad Sci USA 1994 91: 10898–10902

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Tuszynski MH et al. Gene therapy in the adult primate brain: intraparenchymal grafts of cells genetically modified to produce nerve growth factor prevent cholinergic neuronal degeneration Gene Therapy 1996 3: 305–314

    CAS  PubMed  Google Scholar 

  23. Kawaja MD, Rosenberg MB, Yoshida K, Gage FH . Somatic gene transfer of nerve growth factor promotes the survival of axotomized septal neurons and the regeneration of their axons in adult rats J Neurosci 1992 12: 2849–2864

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Blesch A et al. LIF augments corticospinal axon growth and neurotrophin expression after adult CNS injury J Neurosci 1999 19: 3556–3566

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Grill R et al. Cellular delivery of neurotrophin-3 promotes corticospinal axonal growth and partial functional recovery after spinal cord injury J Neurosci 1997 17: 5560–5572

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Tuszynski MH et al. Nerve growth factor delivery by gene transfer induces differential outgrowth of sensory, motor, and noradrenergic neurites after adult spinal cord injury Exp Neurol 1996 137: 157–173

    Article  CAS  PubMed  Google Scholar 

  27. Blesch A, Diergardt N, Uy HS, Tuszynski MH . Cellularly delivered NT-4/5 has axonal growth promoting effects in the injured adult rat spinal cord Soc Neurosci Abstr 1999 25: 497

    Google Scholar 

  28. Weidner N, Blesch A, Grill RJ, Tuszynski MH . Nerve growth factor-hypersecreting Schwann cell grafts augment and guide spinal cord axonal growth and remyelinate central nervous system axons in a phenotypically appropriate manner that correlates with expression of L1 J Comp Neurol 1999 413: 495–506

    Article  CAS  PubMed  Google Scholar 

  29. Harding TC et al. Switching transgene expression in the brain using an adenoviral tetracycline-regulatable system Nat Biotech 1998 16: 553–555

    Article  CAS  Google Scholar 

  30. Corti O et al. Long-term doxycycline-controlled expression of human tyrosine hydroxylase after direct adenovirus-mediated gene transfer to a rat model of Parkinson's disease Proc Nat Acad Sci USA 1999 96: 12120–12125

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Bohl D, Naffakh N, Heard JM . Long-term control of erythropoietin secretion by doxycycline in mice transplanted with engineered primary myoblasts Nat Med 1997 3: 299–305

    Article  CAS  PubMed  Google Scholar 

  32. Tuszynski MH, Gage FH . In vivo assay of neuron-specific effects of nerve growth factor Meth Enzymol 1991 198: 35–48

    Article  CAS  Google Scholar 

  33. Conner JM, Varon S . Nerve growth factor influences the distribution of sympathetic sprouting into the hippocampal formation by implanted superior cervical ganglia Exp Neurol 1994 130: 15–23

    Article  CAS  PubMed  Google Scholar 

  34. Tuszynski MH, U HS, Amaral DG, Gage FH . Nerve growth factor infusion in the primate brain reduces lesion-induced cholinergic neuronal degeneration J Neurosci 1990 10: 3604–3614

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the Paralysis Project, Hollfelder Foundation, Stein Institute for Research on Aging, Alzheimer's Association, Veterans Affairs, National Institute of Neurological Disorders and Stroke (NS37083) and National Institute for Aging (AG10435).

Author information

Authors and Affiliations

  1. Department of Neurosciences-0626, University of California, San Diego, La Jolla, CA, USA

    A Blesch, JM Conner & MH Tuszynski

  2. Veterans Administration Medical Center, San Diego, CA, USA

    MH Tuszynski

Authors
  1. A Blesch
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. JM Conner
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. MH Tuszynski
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Blesch, A., Conner, J. & Tuszynski, M. Modulation of neuronal survival and axonal growth in vivo by tetracycline-regulated neurotrophin expression. Gene Ther 8, 954–960 (2001). https://doi.org/10.1038/sj.gt.3301480

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/sj.gt.3301480

Keywords

Search

Advanced search

Quick links