Neuroprotection and ERK activation by HSV-2 gene ICP10PK

超出所提供的空间。神经元的死亡（凋亡）发生在急性和慢性神经变性疾病如中风、阿尔茨海默病、帕金森病和亨廷顿病中，并且主要由活化的半胱氨酸蛋白酶（半胱天冬酶）介导。神经元死亡的级联反应逐渐出现，为治疗干预提供了一个有效的时间。然而，神经元损伤的广泛性对治疗策略的发展提出了相当大的挑战。设计了几种策略来中断凋亡级联，但它们受到与毒性或不能保留神经元功能相关的问题的限制，这可能是由于靶向在凋亡级联中后期起作用的效应物。初步研究表明，HSV-2基因（ICP 10 PK）可抑制体外培养的中枢神经系统神经元因各种刺激而发生的凋亡。我们构建了一个生长受损的HSV-2突变体（ICP 10 RR），它具有ICP 10 PK和抗凋亡活性，在纹状体内注射后没有毒性，并在鼻内递送后扩散到CNS（包括海马）中的连接部位。病毒感染细胞的神经保护潜力是由于Raf/MEK/ERK存活途径的激活。我们建议评估ICP 10 PK在体内急性兴奋性毒性损伤中的治疗潜力，并确定抗凋亡活性的机制。具体目标是：（i）在以去除营养生长支持或氧化应激为代表的范例中检查ICP 10 PK抗凋亡活性的机制，（ICP 10 PK）和下游（XIAP或p35）凋亡效应物，并检查它们的抗凋亡活性（相对于ICP 10 RR）在用kianic acid处理的器官型培养物中（iii）为了确定ICP 10 RR和X1 AP/p35突变体在体内预防兴奋性毒性死亡的能力，以及（iv）确定海马中的ICP 10 PK表达是否维持突触传递和功能可塑性。这些研究将为开发用于治疗与细胞凋亡相关的急性和慢性神经退行性疾病的基于ICP 10 PK的疗法提供重要信息。性能现场=

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===========================================