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Amelioration of chronic neuropathic pain after partial nerve injury by adeno-associated viral (AAV) vector-mediated over-expression of BDNF in the rat spinal cord

Gene Therapy volume 9, pages 1387–1395 (2002)Cite this article

Abstract

Changing the levels of neurotrophins in the spinal cord micro-environment after nervous system injury has been proposed to recover normal function, such that behavioral response to peripheral stimuli does not lead to chronic pain. We have investigated the effects of recombinant adeno-associated viral (rAAV)-mediated over-expression of brain-derived neurotrophic factor (BDNF) in the spinal cord on chronic neuropathic pain after unilateral chronic constriction injury (CCI) of the sciatic nerve. The rAAV-BDNF vector was injected into the dorsal horn at the thirteenth thoracic spinal cord vertebra (L1 level) 1 week after CCI. Allodynia and hyperalgesia induced by CCI in the hindpaws were permanently reversed, beginning 1 week after vector injection, compared with a similar injection of a control rAAV-GFP vector (green fluorescent protein) or saline. In situ hybridization for BDNF demonstrated that both dorsal and ventral lumbar spinal neurons contained an intense signal for BDNF mRNA, at 1 to 8 weeks after vector injection. There was no similar BDNF mRNA over-expression associated with either injections of saline or rAAV-GFP. These data suggest that chronic neuropathic pain is sensitive to early spinal BDNF levels after partial nerve injury and that rAAV-mediated gene transfer could potentially be used to reverse chronic pain after nervous system injuries in humans.

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References

  1. Dickenson AH, Chapman V, Green GM . The pharmacology of excitatory and inhibitory amino acid-mediated events in the transmission and modulation of pain in the spinal cord Gen Pharm 1997 28: 633–638

    Article  CAS  Google Scholar 

  2. Kerchner GA, Li P, Zhuo M . Speaking out of turn: a role for silent synapses in pain IUBMB Life 1999 48: 251–256

    Article  CAS  PubMed  Google Scholar 

  3. Kawamata M, Omote K . Involvement of increased excitatory amino acids and intracellular Ca2+ concentration in the spinal dorsal horn in an animal model of neuropathic pain Pain 1996 68: 85–96

    Article  CAS  PubMed  Google Scholar 

  4. Zhang A-L, Hao J-X, Seiger A, Xu X-J . Decreased GABA immunoreactivity in spinal cord dorsal horn neurons after transient spinal cord ischemia in the rat Brain Res 1994 656: 187–190

    Article  CAS  PubMed  Google Scholar 

  5. Kingery WS, Fields RD, Kocsis JD . Diminished dorsal root GABA sensitivity following chronic peripheral nerve injury Exp Neurol 1988 100: 478–490

    Article  CAS  PubMed  Google Scholar 

  6. Deng YS, Zhong JH, Zhou XF . Effects of endogenous neurotrophins on sympathetic sprouting in the dorsal root ganglia and allodynia following spinal nerve injury Exp Neurol 2000 164: 344–350

    Article  CAS  PubMed  Google Scholar 

  7. Zhou XF, Deng YS, Xian CJ, Zhong JH . Neurotrophins from dorsal root ganglia trigger allodynia after spinal nerve injury in rats Eur J Neurosci 2001 12: 100–105

    Article  Google Scholar 

  8. Millan MJ . The induction of pain: an integrative review Prog Neurobiol 1999 57: 1–164

    Article  CAS  PubMed  Google Scholar 

  9. Ohsawa F, Widmer HR, Knusel B, Denton TL . Response of embryonic rat hippocampal neurons in culture to neurotropin-3, brain-derived neurotrophic factor and basic fibroblast factor Neurosci 1993 57: 67–77

    Article  CAS  Google Scholar 

  10. Hyman C, Juhasz M, Jackson C, Wright P . Overlapping and distinct actions of the neurotrophins BDNF,NT-3 and NT-4/5 on cultured dopaminergic and GABAergic neurons of the ventral mesencephalon J Neurosci 1994 14: 335–347

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Martin-Iverson MT, Todd KG, Altar CA . Brain-derived neurotrophic factor and neurotrophin-3 activate striatal dopamine and serotonin metabolism and related behaviors: interactions with amphetamine J Neurosci 1994 14: 1262–1270

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Zhou X, Rush R . Endogenous brain-derived neurotrophic factor is anterogradely transported in primary sensory neurons Neurosci 1996 74: 945–953

    Article  CAS  Google Scholar 

  13. Cho H, Kim J, Park H, Kim D . Changes in brain-derived neurotrophic factor immunoreactivity in rat dorsal root ganglia, spinal cord, and gracile nuclei following cut or crush injuries Exp Neurol 1998 154: 224–230

    Article  CAS  PubMed  Google Scholar 

  14. Hermens WT, Verhaagen J . Viral vectors, tools for gene transfer in the nervous system Prog Neurobiol 1998 55: 399–432

    Article  CAS  PubMed  Google Scholar 

  15. Kaplitt M, Makimura H . Defective viral vectors as agents for gene transfer in the nervous system J Neurosci Meth 1997 71: 125–132

    Article  CAS  Google Scholar 

  16. Bankiewicz K, Leff S, Nagy D, Jungles S . Practical aspects of the development of ex vivo and in vivo gene therapy for parkinson's disease Exp Neurol 1997 144: 147–156

    Article  CAS  PubMed  Google Scholar 

  17. Rolling F, Samulski R . AAV as a viral vector for human gene therapy. Generation of recombinant virus Mol Biotech 1995 3: 9–15

    Article  CAS  Google Scholar 

  18. Peel MJ, Zolotukhin S, Schrimsher GW, Muzyczka N . Efficient transduction of green fluorescence protein in spinal cord neurons using adeno-associated virus vectors containing cell type specific promoters Gene Therapy 1997 4: 16–24

    Article  CAS  PubMed  Google Scholar 

  19. Kaplitt M, Leone P, Samulski R, Xiao X . Long-term gene expression and phenotypic correction using adeno-associated virus vectors in the mammalian brain Nat Gen 1994 8: 148–154

    Article  CAS  Google Scholar 

  20. McCown T, Xiao X, Li J, Breese G . Differential and persistent expression patterns of CNS gene transfer by an adeno-associated virus (AAV) vector Brain Res 1996 713: 99–107

    Article  CAS  PubMed  Google Scholar 

  21. Holtmaat A, Hermens W, Sonnemans M, Giger R . Adenoviral vector-mediated expression of B-50/GAP-43 induces alterations in the membrane organization of olfactory axon terminals in vivo J Neurosci 1997 17: 6575–6586

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Bueler H . Adeno-associated viral vectors for gene transfer and gene therapy Biol Chem 1999 380: 613–622

    Article  CAS  PubMed  Google Scholar 

  23. Peel AL, Klein RL . Adeno-associated virus vectors: activity and applications in the CNS J Neurosci Meth 2000 98: 95–104

    Article  CAS  Google Scholar 

  24. Klein RL, Meyer EM, Peel AL, Zolotukhin S . Neuron-specific transduction in the rat septohippocampal or nigrostriatal pathway by recombinant adeno-associated virus vectors Exp Neurol 1998 150: 183–194

    Article  CAS  PubMed  Google Scholar 

  25. Klein RL, Lewis MH, Muzyczka N, Meyer EM . Prevention of 6-hydroxydopamine-indiced rotational behavior by BDNF somatic gene transfer Brain Res 1999 847: 314–320

    Article  CAS  PubMed  Google Scholar 

  26. Loeb JE, Cordier WS, Harris ME, Weitzman MD . Enhanced expression of transgenes from adeno-associated virus vectors with the woodchuck hepatitis virus posttranscriptional regulatory element: implications for gene therapy Hum Gene Ther 1999 10: 2295–2305

    Article  CAS  PubMed  Google Scholar 

  27. Paterna JC, Moccetti T, Mura A, Feldon J . Influence of promoter and WHV post-transcriptional regulatory element on AAV-mediated transgene expression in the rat brain Gene Therapy 2000 7: 1304–1311

    Article  CAS  PubMed  Google Scholar 

  28. Paterna JC, Moccetti T, Mura A, Feldon J . Influence of promoter and WHV post-transcriptional regulatory element on AAV-mediated transgene expression in the rat brain Oudega M et al. Amelioration of chronic neuropathic pain by adeno-associated viral (AAV) vector-mediated overexpression of BDNF in the rat spinal cord. Soc Neurosci Abstr 2000; 26: 169a.

  29. Arenas E, Akerud P, Wong V, Boylan C . Effects of BDNF and NT4/5 on striatonigral neuropeptides or nigral GABA neurons in vivo Eur J Neurosci 1996 8: 1707–1717

    Article  CAS  PubMed  Google Scholar 

  30. Larkfors L, Lindsay RM, Alderson RF . Characterization of the responses of Purkinje cells to neurotrophin treatment J Neurochem 1996 66: 1362–1373

    Article  CAS  PubMed  Google Scholar 

  31. Mizuno K, Carnahan J, Nawa H . Brain-derived neurotrophic factor promotes differentiation of striatal GABAergic neurons Dev Biol 1994 165: 243–256

    Article  CAS  PubMed  Google Scholar 

  32. Spenger C, Hyman C, Studer L, Egli M . Effects of BDNF on dopaminergic, serotonergic and GABA ergic neurons in cultures of human fetal ventral mesencephalon Exp Neurol 1995 133: 50–63

    Article  CAS  PubMed  Google Scholar 

  33. Siuciak JA, Altar CA, Wiegand SJ, Lindsay RM . Antinociceptive effect of brain-derived neurotrophic factor and neurotrophin-3 Brain Res 1994 63: 326–330

    Article  Google Scholar 

  34. Xu XM, Guenard V, Kleitman N, Bunge MB . A combination of BDNF and NT-3 promotes supraspinal axonal regeneration into Schwann cell grafts in adult rat thoracic spinal cord Dev Biol 1995 168: 124–137

    Article  Google Scholar 

  35. Eaton MJ, Plunkett JA, Martinez MA, Karmally S . Changes in GAD and GABA immunoreactivity in the spinal dorsal horn after peripheral nerve injury and promotion of recovery by lumbar transplant of immortalized serotonergic neurons J Chem Neuroanat 1998 16: 57–72

    Article  CAS  PubMed  Google Scholar 

  36. Olson L . Grafts and growth factors in CNS. Basic science with clinical promise Stereo Func Neurosurg 1990 54: 250–267

    Article  Google Scholar 

  37. Hao J-X, Xu X-J, Aldskogius H, Seiger A . Chronic pain related syndrome in rats after ischemic spinal cord lesion: a possible animal model for pain in patients with spinal cord injury Pain 1992 48: 279–290

    Article  PubMed  Google Scholar 

  38. Yaksh TL . Behavioral and autonomic correlates of the tactile evoked allodynia produced by spinal glycine inhibition: effects of modulatory receptor systems and excitatory amino acid antagonists Pain 1989 37: 111–123

    Article  CAS  PubMed  Google Scholar 

  39. Stiller CO, Cui JG, O'Connor WT, Brodin EG . ABA-ergic mechanisms may be involved in the spinal cord stimulation induced reversion of allodynia: animal studies with microdialysis Am Pain Soc Abs 1995 95: 801

    Google Scholar 

  40. Kjaer M, Nielsen H . The analgesic effect of the GABA-agonist THIP in patients with chronic pain of malignant origin Br J Clin Pharm 1983 16: 477–485

    Article  CAS  Google Scholar 

  41. Klein R . Role of mouse neurotrophins in mouse neuronal development FASEB J 1994 8: 738–744

    Article  CAS  PubMed  Google Scholar 

  42. Koliatsos VE, Clatterbuck RE, Winslow JW, Cayouette MH . Evidence that brain-derived neurotrophic factor is a trophic factor for motor neurons in vivo Neuron 1993 10: 359–367

    Article  CAS  PubMed  Google Scholar 

  43. McMahon SB, Armanini MP, Ling L, Phillips HS . Expression and coexpression of trk receptors in subpopulations of adult primary sensory neurons projecting to identified peripheral targets Neuron 1994 12: 1161–1171

    Article  CAS  PubMed  Google Scholar 

  44. Schectersom L, Bothwell M . Novel roles for neurotrophins are suggested by BDNF and NT-3 mRNA expression in developing neurons Neuron 1992 9: 449–463

    Article  Google Scholar 

  45. Michael G, Averill S, Nitkunan A, Rattray M . Nerve growth factor treatment increases brain-derived neurotrophic factor selectively in TrkA-expressing dorsal root ganglion cells and in their central terminations within the spinal cord J Neurosci 1997 17: 8476–8490

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Griesbeck O, Canossa M, Campana G, Gartner A . Are there differences between secretion characteristics of NGF and BDNF? Implications for the modulatory role of neurotrophins in activity-dependent neuronal plasticity Microsc Res Tech 1999 45: 262–275

    Article  CAS  PubMed  Google Scholar 

  47. Thompson S, Bennett D, Kerr B, Bradbury E . Brain-derived neurotrophic factor is an endogenous modulator of nociceptive responses in the spinal cord Proc Nat Acad Sci USA 1999 96: 7714–7718

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  48. Kerr B, Bradbury E, Bennet D, Trivedi P . Brain-derived neurotropic factor modulates nociceptive sensory inputs and NMDA-evoked responses in the rat spinal cord J Neurosci 1999 19: 5138–5148

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  49. Rutherford LC, Nelson S, Turrigiano G . BDNF has opposite effects on the quantal amplitude of pyramidal neuron and interneuron excitatory synapses Neuron 1998 21: 521–530

    Article  CAS  PubMed  Google Scholar 

  50. Oyelese A, Rizzo M, Waxman S, Kocsis J . Differential effects of NGF and BDNF on axotomy-induced changes in GABA (A)-receptor-mediated conductance and sodium currents in cutaneous afferent neurons J Neurophysiology 1997 78: 31–42

    Article  CAS  Google Scholar 

  51. Frank L, Wiegand SJ, Siuciak JA, Lindsay RM . Effects of BDNF infusion on the regulation of TrkB protein and message in adult rat brain Exp Neurol 1997 145: 62–70

    Article  CAS  PubMed  Google Scholar 

  52. Eaton MJ, Staley JK, Globus MYT, Whittemore SR . Developmental regulation of early serotonergic neuronal differentiation: the role of brain-derived neurotrophic factor and membrane depolarization Dev Biol 1995 170: 169–182

    Article  CAS  PubMed  Google Scholar 

  53. Eaton MJ, Dancausse HR, Santiago DI, Whittemore SR . Lumbar transplants of immortalized serotonergic neurons alleviates chronic neuropathic pain Pain 1997 72: 59–69

    Article  CAS  PubMed  Google Scholar 

  54. Hains BC, Johnson KM, McAdoo DJ, Eaton MJ . Engraftment of immortalized serotonergic neurons enhances locomotor function and attenuates pain-like behavior following spinal hemisection injury in the rat Exp Neurol 2001 171: 361–378

    Article  CAS  PubMed  Google Scholar 

  55. Hains BC, Fullwood SD, Eaton MJ, Hulsebosch CE . Subdural engraftment of serotonergic neurons following spinal hemisection restores spinal serotonin, downregulates the serotonin transporter, and increases BDNF tissue content in the rat Brain Res 2001 913: 35–46

    Article  CAS  PubMed  Google Scholar 

  56. Yaksh TL, Wilson PR . Spinal serotonin terminal system mediates antinociception J Pharm Exp Ther 1979 208: 446–453

    CAS  Google Scholar 

  57. Davies JE . Roberts MHT. 5-hydroxytryptamine reduces substance P responses on dorsal interneurones: a possible interaction of neurotransmitters Brain Res 1981 217: 399–404

    Article  CAS  PubMed  Google Scholar 

  58. Anden NE, Jukes MGM, Lundberg A . Spinal reflexes and monoamine liberation Nature 1964 202: 1222–1223

    Article  CAS  PubMed  Google Scholar 

  59. Anden NE, Jukes MGM, Lundberg A . Spinal reflexes and monoamine liberation Besson J-M. Serotonin and Pain. Excerpta Medica: New York, 1990.

  60. Millan MJ . Serotonin and pain: evidence that activation of 5-HT1A receptors does not elicit antinociception against noxious thermal, mechanical and chemical stimuli in mice Pain 1994 58: 45–61

    Article  CAS  PubMed  Google Scholar 

  61. Carter JE, Schuchman EH . Gene therapy for neurodegenerative diseases: fact or fiction? Br J Psychiatry 2001 178: 392–394

    Article  CAS  PubMed  Google Scholar 

  62. Crameri A, Whitehorn EA, Tate E, Stemmer WP . Improved green fluorescent protein by molecular evolution using DNA shuffling Nat Biotechnol 1996 14: 315–319

    Article  CAS  PubMed  Google Scholar 

  63. Zolotukhin S, Potter M, Hauswirth WW, Guy J . A ‘humanized’ green fluorescent protein cDNA adapted for high-level expression in mammalian cells J Virol 1996 70: 4646–4654

    CAS  PubMed  PubMed Central  Google Scholar 

  64. Donello JE, Loeb JE, Hope TJ . Woodchuck hepatitis virus contains a tripartite posttranscriptional regulatory element J Virol 1998 72: 5085–5092

    CAS  PubMed  PubMed Central  Google Scholar 

  65. Grimm D, Kern A, Rittner K, Kleinschmidt JA . Novel tools for production and purification of recombinant adeno associated virus vectors Hum Gene Ther 1998 9: 2745–2760

    Article  CAS  PubMed  Google Scholar 

  66. Hermens WT, ter Brake O, Dijkhuizen PA, Sonnemans MA . Purification of recombinant adeno-associated virus by iodixanol gradient ultracentrifugation allows rapid and reproducible preparation of vector stocks for gene transfer in the nervous system Hum Gene Ther 1999 10: 1885–1891

    Article  CAS  PubMed  Google Scholar 

  67. Salvetti A, Oreve S, Chadeuf G, Favre D . Factors influencing recombinant adeno-associated virus production Hum Gene Ther 1998 9: 695–706

    Article  CAS  PubMed  Google Scholar 

  68. Bennett GJ, Xie YK . A peripheral mononeuropathy in rat that produces disorders of pain sensation like those seen in man Pain 1988 33: 87–107

    Article  CAS  PubMed  Google Scholar 

  69. Cejas PJ, Martinez M, Karmally S, McKillop M . Lumbar transplant of neurons genetically modified to secrete brain-derived neurotrophic factor attenuate allodynia and hyperalgesia after sciatic nerve constriction Pain 2000 86: 195–210

    Article  CAS  PubMed  Google Scholar 

  70. Zimmermann M . Ethical guidelines for investigations of experimental pain in conscious animals Pain 1983 16: 109–110

    Article  CAS  PubMed  Google Scholar 

  71. Schaeren-Wiemers N, Gerfin-Moser A . A single protocol to detect transcripts of various types and expression levels in neural tissue and cultured cells: in situ hybridization using digoxigenin-labeled cRNA probes Histochem 1993 100: 431–440

    Article  CAS  Google Scholar 

  72. Giger RJ, Wolfer DP, De Wit GM, Verhaagen J . Anatomy of rat semaphorin III/collapsin-1 mRNA expression and relationship to developing nerve tracts during neuroembryogenesis J Comp Nerol 1996 375: 378–392

    Article  CAS  Google Scholar 

  73. Pasterkamp RJ, Giger RJ, Ruitenberg MJ, Holtmaat A . Expression of the gene encoding the chemorepellent semaphorin III is induced in the fibroblast component of neural scar tissue formed following injuries of adult but not neonatal CNS Mol Cell Neurosci 1999 13: 143–166

    Article  CAS  PubMed  Google Scholar 

  74. Hargreaves K, Dubner R, Brown F, Flores C . A new and sensitive method for measuring thermal nociception in cutaneous hyperalgesia Pain 1988 32: 77–88

    Article  CAS  PubMed  Google Scholar 

  75. Chaplan SR, Bach FW, Pogrel JW, Chung JM . Quantitative assessment of tactile allodynia in the rat paw J Neurosci Meth 1994 53: 55–63

    Article  CAS  Google Scholar 

  76. Randall L, Selitto J . A method for measurement of analgesic activity on inflamed tissue Arch Int Pharmacodyn 1957 4: 409–419

    Google Scholar 

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Acknowledgements

The authors would like to thank Dr Beata Frydel, Director of the Image Analysis Facility, Mr Miguel Martinez, and Mr Jose Lorenzo, of the Miami Project to Cure Paralysis, for their excellent technical assistance. The work done in the laboratories of Dr M Eaton was supported by the Miami Project to Cure Paralysis and The State of Florida. Work performed in the laboratories of Dr J Verhaagen was supported by a Pioneer grant from the Netherlands Organization for Scientific Research.

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Authors and Affiliations

  1. The Miami Project To Cure Paralysis, University of Miami School of Medicine, Miami, FL, USA

    M J Eaton, B Blits & M Oudega

  2. Department of Neurological Surgery, University of Miami School of Medicine, Miami, FL, USA

    M J Eaton & M Oudega

  3. VA RR&D Center of Excellence in Functional Recovery in Chronic Spinal Cord Injury, VAMC, Miami, FL, USA

    M J Eaton

  4. Graduate School for Neurosciences Amsterdam, Netherlands Institute for Brain Research, Amsterdam, The Netherlands

    B Blits, M J Ruitenberg & J Verhaagen

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Eaton, M., Blits, B., Ruitenberg, M. et al. Amelioration of chronic neuropathic pain after partial nerve injury by adeno-associated viral (AAV) vector-mediated over-expression of BDNF in the rat spinal cord. Gene Ther 9, 1387–1395 (2002). https://doi.org/10.1038/sj.gt.3301814

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