Neuroprotection and ERK activation by HSV-2 gene ICP10PK

超出提供的空间。神经元死亡(细胞凋亡)发生在急性和慢性神经退行性疾病中，如中风、阿尔茨海默病、帕金森病和亨廷顿病，主要由激活的半胱氨酸蛋白酶(Caspase)介导。神经元死亡的级联逐渐出现，为治疗干预提供了一个有效的时期。然而，神经元损伤的广泛性给治疗策略的发展提出了一个不可忽视的挑战。人们设计了几种策略来阻断凋亡级联反应，但它们都受到毒性或未能保留神经元功能的问题的限制，这可能是由于靶向作用于在凋亡级联反应中较晚发挥作用的效应器。我们的最新数据表明，HSV-2基因(ICP10PK)可防止各种刺激诱导的体外中枢神经系统神经元的凋亡。我们构建了一个生长受损的HSV-2突变体(ICP10 RR)，它保持了ICP10PK和抗凋亡活性，在纹状体注射后无毒，并在鼻腔给药时扩散到中枢神经系统(包括海马区)的连接部位。病毒感染细胞的神经保护作用是由于激活了Raf/MEK/ERK生存通路。我们建议评估ICP10 PK在体内急性兴奋性毒性损伤中的治疗潜力，并确定其抗凋亡活性的机制。这些具体目标是：(1)检测以去除营养生长支持或氧化应激为代表的范例中ICP10 PK的抗凋亡活性的机制；(2)构建同时针对上游(ICP10 PK)和下游(XIAP或p35)的凋亡效应载体，并在含有奎尼酸的器官型培养物中检测它们(相对于ICP10 RR)的抗凋亡活性；(3)确定ICP10 RR和XlAP/p35突变体在体内防止兴奋性死亡的能力；以及(Iv)确定ICP10PK在海马区的表达是否维持突触传递和功能可塑性。这些研究将为开发基于ICP10PK的治疗与细胞凋亡相关的急慢性神经退行性疾病提供所需的重要信息。表演网站========================================Section End===========================================

EXCEED THE SPACE PROVIDED. 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. PERFORMANCE SITE ========================================Section End===========================================