Herpes simplex virus mediated nerve growth factor expression in bladder and afferent neurons: Potential treatment for diabetic bladder dysfunction
Herpes simplex virus mediated nerve growth factor expression in bladder and afferent neurons: Potential treatment for diabetic bladder dysfunction
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DOI：
10.1016/s0022-5347(05)66407-5
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发表时间：
2001-05-01
影响因子：
6.6
通讯作者：
Chancellor, MB
中科院分区：
文献类型：
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作者：
Goins, WF;Yoshimura, N;Chancellor, MB
目的：由感觉神经病变引起的糖尿病性膀胱病变有可能通过直接基因治疗。研究表明，神经生长因子（NGF）对糖尿病患者控制膀胱功能的传出背根神经节神经元的死亡具有改善作用。我们研究了神经生长因子基因转移到膀胱和膀胱传入通路治疗糖尿病性膀胱病变。我们使用具有复制能力和复制缺陷的1型单纯疱疹病毒（HSV-1）载体，表达小鼠NGF的β -亚基（β -NGF）的功能活性形式，以检测给药载体后治疗性基因表达的水平和持续时间。材料与方法：将1 × 10(6) ~ 1 × 10(8) pfu HSV-NGF表达载体注入成年大鼠膀胱壁后，采用酶联免疫吸附试验（ELISA）和免疫组化检测NGF在急性感染（3 d）和潜伏感染（21 d）期间的表达情况。结果：用强人巨细胞病毒即时早期启动子驱动β -NGF基因表达的HSV载体在感染后3天膀胱和膀胱传入神经元所在的L6 ~ S1背根神经节中NGF升高，ELISA分析显示膀胱组织和背rest神经节中的NGF分别比对照载体增加7 ~ 9倍和2 ~ 4倍。抗NGF抗体免疫组化染色也检测到L6 ~ S1背根神经节神经元NGF表达升高。注射后21 d， ELISA检测延长的NGF表达。含有HSV-1潜伏期启动子（LAP-2）驱动NGF的复制缺陷载体在膀胱注射后21天在膀胱和背根神经节表达NGF。ELISA分析证实，膀胱和L6至S1背根神经节中NGF的表达增加了约2至3倍。结论：NGF基因可通过HSV载体在膀胱和膀胱传入通路中转移表达。据我们所知，我们的研究首次证明了基因治疗改变内脏感觉神经元神经营养表达的有效性。这种基因转移技术可能有助于治疗某些类型的神经源性膀胱功能障碍，如糖尿病性膀胱病变，其中NGF转运减少可能是一个致病因素。
Purpose: Diabetic cystopathy resulting from sensory neuropathy may potentially be treated by direct gene therapy. It has been suggested that nerve growth factor (NGF) has an ameliorative effect in preventing the death in diabetes of efferent dorsal root ganglion neurons, which control bladder function. We investigated NGF gene transfer to the bladder and bladder afferent pathways for treating diabetic cystopathy. We used replication competent and replication defective herpes simplex virus type 1 (HSV-1) vectors that express a functionally active form of the beta -subunit of mouse NGF (beta -NGF) to examine the level and duration of therapeutic, gene expression after administration of the vectors.Materials and Methods: NGF expression during acute (3 days) and latent (21 days) infections was assessed by enzyme-linked immunosorbent assay (ELISA) and immunohistochemical testing after the injection of 1 x 10(6) to 1 x 10(8) pfu HSV-NGF expression vectors into the bladder wall of adult rats.Results: HSV vectors with the strong human cytomegalovirus immediate early promoter used to drive beta -NGF gene expression exhibited increased NGF 3 days after infection in the bladder and L6 to S1 dorsal root ganglia, where bladder afferent neurons are located, ELISA analysis revealed that NGF in the bladder tissue and dorsal rest ganglia was increased 7 to 9 and 2 to 4-fold, respectively, over the control vector. Increased NGF expression in L6 to S1 dorsal root ganglia neurons was also detected by immunohistochemical staining with antiNGF antibodies. Extended NGF expression was detected by ELISA 21 days after injection. Replication defective vectors containing HSV-1 latency promoter (LAP-2) driving NGF expressed NGF in the bladder and dorsal root ganglia 21 days after bladder injection. ELISA analysis confirmed an approximate 2 to 3-fold increase of NGF expression in the bladder and L6 to S1 dorsal 1 root ganglia.Conclusions: The NGF gene may be transferred and expressed in the bladder and bladder afferent pathways using HSV vectors. To our knowledge our study represents the first demonstration of the effectiveness of gene therapy for altering neurotrophic expression in visceral sensory neurons. This technique of gene transfer may be useful for treating certain types of neurogenic bladder dysfunction, such as, diabetic cystopathy, in which decreased NGF transport may be a causative factor.