Role of 14-3-3 Proteins in Alpha-Synuclein-Induced Neurotoxicity

描述(由申请人提供)：α-突触核蛋白在帕金森氏病的发病机制中起着核心作用，但它如何触发多巴胺能神经元毒性尚不清楚。最近对转基因α-突触核蛋白小鼠的基因芯片研究表明，α-突触核蛋白的过度表达会导致一类编码14-3-3蛋白的基因表达下降。由于14-3-3S调控包括细胞凋亡在内的关键信号级联反应，14-3-3S的减少可能是α-突触核蛋白诱导的毒性的重要一步。候选人已经获得了支持14-3-3蛋白，特别是theta异构体的神经保护潜力的初步数据。拟议的实验将确定在帕金森病的体外和体内模型中，14-3-3亚型可以在多大程度上减轻α-突触核蛋白的毒性，以及它们是否可以降低这些模型中的凋亡因子活性。α-突触核蛋白如何导致14-3-3表达减少的潜在机制将被研究。评估磷酸化在14-3-3S和α-突触核蛋白相互作用中的作用的研究也将被进行。这些研究的结果将验证14-3-3蛋白作为帕金森病和相关疾病治疗的潜在靶点。拟议职业发展计划的研究部分将加深候选人对帕金森氏病病理生理学的理解，以及她在实验技能方面的培训，如体内动物工作、表观遗传学方法和使用病毒载体进行基因传递。她将继续接受帕金森氏症和相关疾病的诊断和治疗方面的专科培训和临床实践。此外，候选人将在临床研究培训中进行正式研究，以获得关于将动物模型中的神经保护分子转化为人类患者治疗的主要问题的知识。到五年计划结束时，候选人预计将完全准备好指导帕金森氏病机制的研究，并将此类研究转化为潜在的治疗方法。14-3-3蛋白质看起来很有希望，但无论它们最终是否成为实用的神经保护剂，研究这些蛋白质获得的经验将教会候选人如何测试候选蛋白质是否真的具有保护性，如果是的话，如何开始将它们开发成临床治疗。

DESCRIPTION (provided by applicant): Alpha-synuclein has a central role in the pathogenesis of Parkinson's disease, yet how it triggers dopaminergic neuron toxicity is poorly understood. Recent gene microarray studies in transgenic alpha-synuclein mice have shown that over-expression of alpha-synuclein results in decreased expression of a class of genes encoding the 14-3-3 proteins. Because the 14-3-3s regulate key signaling cascades, including apoptosis, the decrease in 14-3-3s may be an important step in alpha-synuclein-induced toxicity. The candidate has obtained preliminary data supporting the neuroprotective potential of the 14-3-3 proteins, particularly the theta isofom. Proposed experiments will determine the extent to which 14-3-3 isoforms can mitigate alpha-synuclein toxicity in both in vitro and in vivo models of Parkinson's disease and whether they can reduce apoptotic factor activity in these models. Potential mechanisms of how alpha-synuclein causes decreased 14-3-3 expression will be examined. Studies to evaluate the role of phosphorylation in the interaction between 14-3-3s and alpha-synuclein will also be performed. Results from these studies will look to validate the 14-3-3 proteins as potential targets for therapy in Parkinson's disease and related disorders. The research portion of the proposed career development program will further the candidate's understanding of Parkinson's disease pathophysiology and her training in experimental skills, such as in vivo animal work, epigenetic methods and the use of viral vectors for gene delivery. She will continue her subspecialty training and clinical practice in the diagnosis and treatment of Parkinson's disease and related disorders. In addition, the candidate will pursue formal studies in clinical research training to acquire knowledge on the major issues for converting a molecule neuroprotective in animal models into treatment for human patients. By the end of the five-year plan, the candidate expects to be fully equipped to direct research on Parkinson's disease mechanisms and the translation of such research into potential therapies. The 14-3-3 proteins appear promising, but whether or not they turn out to become practical neuroprotective agents, the experience gained in studying these proteins will have taught the candidate how to test whether candidate proteins are truly protective and, if so, how to begin developing them into clinical therapy.