Neuroprotection and ERK activation by HSV-2 gene ICP10PK

神经元死亡(细胞凋亡)在急性和慢性神经退行性疾病中都会发生，例如 中风、阿尔茨海默氏症、帕金森氏症和亨廷顿氏病，主要由激活的 半胱氨酸蛋白酶(Caspase)。神经元死亡的级联逐渐出现，提供了一段时间 _可用于治疗干预。然而，神经元损伤的广泛性表现为 ‘，对治疗策略的发展提出了不可忽视的挑战。设计了几种策略来 _终止细胞凋亡级联反应，但受限于毒性或未能保留 神经元的功能，可能是由于靶向的效应，功能后期的凋亡级联。我们的 最新数据表明，HSV-2基因(ICP10PK)可在体外阻止中枢神经系统神经元的凋亡 由各种刺激引起的。我们构建了一个生长受限的HSV-2突变体(ICP10 RR)， 具有ICP10PK和抗凋亡活性，纹状体内注射后无毒，并传播到 鼻腔给药时中枢神经系统(包括海马体)的连接部位。神经保护潜能 病毒感染细胞是由于激活了Raf/MEK/ERK生存通路。我们建议评估 ICP10PK在急性兴奋性毒性损伤中的体内治疗作用及机制探讨 抗细胞凋亡活性。具体目的是：(1)研究ICP10 PK抗细胞凋亡的机制 以去除营养生长支持或氧化应激为代表的范例中的活性，(Ii)工程 同时针对上游(ICP10 Pk)和下游(XIAP或p35)凋亡效应因子的载体 并检测它们的抗凋亡性(相对于ICP10 RR)，并在含有卡拉胶的器官型培养中进行检测 ACID(兴奋性毒性模型)，(III)测定ICP10 RR和XlAP/p35突变体防止 以及(Iv)测定ICP10PK在海马区的表达 维持突触传递和功能可塑性。这些研究将提供重要的信息 开发基于ICP10PK的治疗急慢性疾病所需的药物 与细胞凋亡相关的神经退行性疾病。

Death of neurons (apoptosis) occurs in both acute and chronic neurodegenerative diseases such as stroke, Alzheimer's, Parkinson's, and Huntington's diseases and is primarily mediated by activated cysteine proteases (caspases). Cascades of neuronal death emerge gradually, providing a period _vailable for therapeutic intervention. However, the widespread nature of the neuronal injury presents a ',onsiderable challenge to the development of therapeutic strategies. Several strategies were designed to _terrupt the apoptotic cascade but they are limited by problems related to toxicity or the failure to retain neuronal function, likely due to targeting of effectors that function late in the apoptotic cascade. Our )reliminary data indicate that a HSV-2 gene (ICP10PK) prevents apoptosis of CNS neurons in vitro rlduced by various stimuli. We constructed a growth-compromised HSV-2 mutant (ICP10 RR) that etains ICP10PK and anti-apoptotic activity, is not toxic following intrastriatal injection and disseminates to ;onnected sites in the CNS (including hippocampus) upon intranasal delivery. Neuroprotective potential invirus infected cells is due to activation of the Raf/MEK/ERK survival pathway. We propose to evaluate the therapeutic potential of ICP10 PK in acute excitotoxic injury in vivo and define the mechanism of anti-apoptotic activity. The specific aims are: (i) To examine the mechanism of ICP10 PK anti-apoptotic activity in paradigms represented by removal of trophic growth support or oxidative stress, (ii) To engineer vectors that target both both upstream (ICP10 PK) and downstream (XIAP or p35) apoptotic effectors and examine their anti-apopotic activity (relative to ICP10 RR) in organotypic cultures teated with kianic acid (excitotoxic model), (iii) To determine the ability of ICP10 RR and the XlAP/p35 mutants to prevent excitotoxic death in vivo, and (iv) To determine whether ICP10PK expression in the hippocampus maintains synaptic transmission and functional plasticity. The studies will provide significant information required for the development of ICP10PK based therapies for the treatment of acute and chronic neurodegenerative diseases that are associated with apoptosis.