Defective HSV-1 vector expressing BDNF in auditory ganglia elicits neurite outgrowth: model for treatment of neuron loss following cochlear degeneration.

The neurotrophins are a family of growth factors that play an important role in the development and maintenance of the nervous system. Brain-derived neurotrophic factor (BDNF) is a member of the neurotrophin family that appears to participate in the maturation and function of mammalian auditory neurons. Forms of deafness due to varied injurious stimuli that are amenable to treatment with implantable prosthetic devices require the survival of these BDNF-responsive auditory neurons for effective outcome. To evaluate the feasibility of developing a gene therapy for deafness that may be used in conjunction with a prosthetic device, we constructed replication-defective herpes simplex virus (HSV) amplicon vectors that carry the human BDNF cDNA. Using these vectors, HSVbdnf and HSVbdnflac (expresses BDNF and Escherichia coli beta-galactosidase), we evaluated the expression and biological activity in established cell lines and explant cultures prepared from spiral ganglia of the murine ear. Gene transfer with HSVbdnf resulted in the efficient expression of human BDNF mRNA in murine fibroblasts. Using two BDNF-responsive cell lines, PC12trkB and MG87trkB, we demonstrate efficient secretion of biologically active BDNF. Finally, transduction of explanted spiral ganglia with HSVbdnflac elicited robust neuritic process outgrowth comparable to exogenously added BDNF. Overall, these data demonstrate that HSV vectors can efficiently transfer and express the BDNF gene in many cell types, including auditory neurons. Moreover, they suggest that similar vectors may be used to express the neurotrophin in auditory neurons in vivo and perhaps as adjunctive gene therapy for deafness.

[1]  M. Saarma,et al.  Brain-derived neurotrophic factor and neurotrophin 3 mRNAs in the peripheral target fields of developing inner ear ganglia. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[2]  T. R. Van De Water,et al.  The effect of target tissues on survival and differentiation of mammalian statoacoustic ganglion neurons in organ culture. , 1987, Acta Oto-Laryngologica.

[3]  E. Castrén,et al.  Light regulates expression of brain-derived neurotrophic factor mRNA in rat visual cortex. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[4]  D. Goeddel,et al.  Neurotrophin-5: A novel neurotrophic factor that activates trk and trkB , 1991, Neuron.

[5]  G. Yancopoulos,et al.  BDNF is a neurotrophic factor for dopaminergic neurons of the substantia nigra , 1991, Nature.

[6]  P. Chomczyński,et al.  Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. , 1987, Analytical biochemistry.

[7]  Molly Webster,et al.  Spiral ganglion neuron loss following organ of corti loss: A quantitative study , 1981, Brain Research.

[8]  R. Espinosa,et al.  Mammalian neurotrophin-4: structure, chromosomal localization, tissue distribution, and receptor specificity. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[9]  H. Federoff,et al.  Herpes simplex virus type 1 amplicon vectors with glucocorticoid-inducible gene expression. , 1995, Human gene therapy.

[10]  I. Fariñas,et al.  Severe sensory and sympathetic deficits in mice lacking neurotrophin-3 , 1994, Nature.

[11]  Nancy Y. Ip,et al.  Potentiation of developing neuromuscular synapses by the neurotrophins NT-3 and BDNF , 1993, Nature.

[12]  P. Bergold,et al.  Localized gene transfer into organotypic hippocampal slice cultures and acute hippocampal slices , 1993, Journal of Neuroscience Methods.

[13]  J. Bryan,et al.  An efficient deletion mutant packaging system for defective herpes simplex virus vectors: potential applications to human gene therapy and neuronal physiology. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[14]  J. Aran,et al.  Spiral ganglion changes after massive aminoglycoside treatment in the guinea pig. Counts and ultrastructure. , 1982, Acta oto-laryngologica.

[15]  R. Klein.  Role of neurotrophins in mouse neuronal development , 1994, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[16]  R. Everett,et al.  A prominent serine-rich region in Vmw175, the major transcriptional regulator protein of herpes simplex virus type 1, is not essential for virus growth in tissue culture. , 1990, The Journal of general virology.

[17]  D. Boothman,et al.  Expression of the E. coli Lac Z gene from a defective HSV‐1 vector in various human normal, cancer‐prone and tumor cells , 1989, FEBS letters.

[18]  H. Thoenen,et al.  Neurotrophin-6 is a new member of the nerve growth factor family , 1994, Nature.

[19]  D. Choi,et al.  Potentiated necrosis of cultured cortical neurons by neurotrophins. , 1995, Science.

[20]  M. Saarma,et al.  Coordinated expression and function of neurotrophins and their receptors in the rat inner ear during target innervation , 1994, Hearing Research.

[21]  Richard R. Fay,et al.  Clinical Aspects of Hearing , 1996, Springer Handbook of Auditory Research.

[22]  M. Saarma,et al.  Expression patterns of neurotrophin and their receptor mRNAs in the rat inner ear , 1993, Hearing Research.

[23]  M. Goldfarb,et al.  Growth factor requirements of oncogene-transformed NIH 3T3 and BALB/c 3T3 cells cultured in defined media. , 1986, Molecular and cellular biology.

[24]  L. Belluscio,et al.  NT-3, BDNF, and NGF in the developing rat nervous system: Parallel as well as reciprocal patterns of expression , 1990, Neuron.

[25]  J. Nichols,et al.  Dicistronic targeting constructs: reporters and modifiers of mammalian gene expression. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[26]  F. Hefti,et al.  Brain-derived neurotrophic factor administration protects basal forebrain cholinergic but not nigral dopaminergic neurons from degenerative changes after axotomy in the adult rat brain , 1992, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[27]  G. Moonen,et al.  Peripheral and central target-derived trophic factor(s) effects on auditory neurons , 1992, Hearing Research.

[28]  J. Aran,et al.  Progressive degeneration in the cochlear nucleus after chemical destruction of the cochlea. , 1986, Acta oto-laryngologica.

[29]  R. Romand,et al.  Neurotrophins and the development of cochlear innervation. , 1994, Life sciences.

[30]  M. Bothwell,et al.  Neurotrophin and neurotrophin receptor mRNA expression in developing inner ear , 1994, Hearing Research.

[31]  Y. Barde,et al.  Identification and characterization of a novel member of the nerve growth factor/brain-derived neurotrophic factor family , 1990, Nature.

[32]  B. Ryals,et al.  Changes in the acoustic nerve after hair cell regeneration , 1992, Experimental Neurology.

[33]  P. Ernfors,et al.  Cells Expressing mRNA for Neurotrophins and their Receptors During Embryonic Rat Development , 1992, The European journal of neuroscience.

[34]  G. Yancopoulos,et al.  Trkl3 mediates BDNF/NT-3-dependent survival and proliferation in fibroblasts lacking the low affinity NGF receptor , 1991, Cell.

[35]  D. Pfaff,et al.  Preproenkephalin promoter yields region-specific and long-term expression in adult brain after direct in vivo gene transfer via a defective herpes simplex viral vector. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[36]  M. Avila,et al.  Brain-derived neurotrophic factor and neurotrophin-3 support the survival and neuritogenesis response of developing cochleovestibular ganglion neurons , 1993 .

[37]  H. Sobkowicz,et al.  Epithelial repair following mechanical injury of the developing organ of Corti in culture: An electron microscopic and autoradiographic study , 1992, Experimental Neurology.

[38]  Rudolf Jaenisch,et al.  Complementary roles of BDNF and NT-3 in vestibular and auditory development , 1995, Neuron.

[39]  G. Moonen,et al.  Temporal pattern of nerve growth factor (NGF) binding in vivo and the in vitro effects of NGF on cultures of developing auditory and vestibular neurons. , 1991, Acta oto-laryngologica.

[40]  G. Moonen,et al.  Neurotrophins affect survival and neuritogenesis by adult injured auditory neurons in vitro. , 1994, Neuroreport.

[41]  J. T. Weber,et al.  The neurotrophins and their receptors: Structure, function, and neuropathology , 1994, Neuroscience & Biobehavioral Reviews.

[42]  R. Altschuler,et al.  Protective Effect of Electrical Stimulation in the Deafened Guinea Pig Cochlea , 1991, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[43]  J. Wersäll,et al.  Degeneration of neural elements in the cochlea of the guinea-pig after damage to the organ of corti by ototoxic antibiotics. , 1974, Acta oto-laryngologica. Supplementum.

[44]  M. Ard,et al.  Trophic interactions between the cochleovestibular ganglion of the chick embryo and its synaptic targets in culture , 1985, Neuroscience.

[45]  H. Federoff,et al.  Expression of vascular endothelial growth factor from a defective herpes simplex virus type 1 amplicon vector induces angiogenesis in mice. , 1995, Circulation research.

[46]  T. Friedmann,et al.  Cytotoxicity of a replication-defective mutant of herpes simplex virus type 1 , 1992, Journal of virology.

[47]  H. Thoenen,et al.  Placode and neural crest-derived sensory neurons are responsive at early developmental stages to brain-derived neurotrophic factor. , 1985, Developmental biology.