Defective HSV-1 vector expressing BDNF in auditory ganglia elicits neurite outgrowth: Model for treatment of neuron loss following cochlear degeneration
Defective HSV-1 vector expressing BDNF in auditory ganglia elicits neurite outgrowth: Model for treatment of neuron loss following cochlear degeneration
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DOI：
10.1089/hum.1996.7.2-173
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发表时间：
1996-01-20
影响因子：
4.2
通讯作者：
Federoff, HJ
中科院分区：
文献类型：
--
作者：
Geschwind, MD;Hartnick, CJ;Federoff, HJ
神经营养因子是一类在神经系统发育和维持中起重要作用的生长因子。脑源性神经营养因子（BDNF）是神经营养因子家族的一员，参与哺乳动物听觉神经元的成熟和功能。由于各种伤害性刺激而导致的耳聋形式可以用植入式假体装置进行治疗，需要这些BDNF反应性听觉神经元的存活以获得有效的结果。为了评估开发可与假体装置结合使用的耳聋基因治疗的可行性，我们构建了携带人BDNF cDNA的复制缺陷型单纯疱疹病毒（HSV）扩增子载体。使用这些载体，HSVbdnf和HSVbdnflac（表达BDNF和大肠杆菌P-半乳糖苷酶），我们评估了在建立的细胞系和外植体培养制备的小鼠耳螺旋神经节的表达和生物活性。用HSVbdnf基因转染小鼠成纤维细胞，使人BDNF mRNA在细胞内高效表达。使用两个BDNF响应细胞系，PC 12 trkB和MG 87 trkB，我们证明了生物活性BDNF的有效分泌。最后，用HSVbdnflac转导受损的螺旋神经节引起了与外源性添加的BDNF相当的强大的神经炎过程生长。总之，这些数据表明，HSV载体可以有效地转移和表达BDNF基因在许多细胞类型，包括听觉神经元。此外，他们建议类似的载体可以用于在体内听觉神经元中表达神经营养因子，并可能作为耳聋的预防性基因治疗。
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 P-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.