Genetic basis for alpha-synuclein toxicity

描述（由申请人提供）：已知突变和环境因素诱导α-突触核蛋白的各种翻译后改变，其中α-突触核蛋白的改变在帕金森病（PD）的发作和进展中起重要作用。特别是，α-突触核蛋白不溶性和α-突触核蛋白聚集的增加似乎是PD以及PD实验模型中观察到的发病机制和神经毒性的中心事件。衰老是PD和相关突触核蛋白病的主要风险因素，尽管目前尚不清楚与年龄相关的α-突触核蛋白改变的潜在遗传基础。鉴定负性和正性调节有丝分裂后神经元内α-突触核蛋白的年龄相关变化的每个基因可能导致更好地理解PD发病机制的发生和进展的分子基础，以及潜在地导致用于治疗PD的新型疗法的开发。近年来已经鉴定了α-突触核蛋白中的许多突变，观察到α-突触核蛋白中的突变促进了某些家族性形式的PD。与衰老一样，重要的是鉴定在有丝分裂后神经元中与突变的α-突触核蛋白相关的正性和负性调节发病机制和毒性的基因。酿酒酵母（Saccharomyces cerevisiae，S.酿酒酵母）模型系统已被证明对研究细胞衰老是有用的，其中衰老的稳定期模型有助于我们理解有丝分裂后细胞衰老。此外，S.酿酒酵母已经证明在研究许多人类基因的生物学中是有用的，包括与疾病相关的突变人类基因。目前的建议的重点是测试的假设，诱变实验与S。酿酒酵母稳定期老化模型有助于发展对野生型和突变体α-突触核蛋白聚集/不溶性的年龄相关改变以及野生型和突变体α-突触核蛋白毒性的年龄相关增加的遗传基础的理解。研究将在表达野生型和突变型（A53T.A30P）α-突触核蛋白的细胞中进行。具体目的如下：1）利用诱变来理解α-突触核蛋白发生的年龄相关改变的遗传基础2）利用诱变来理解α-突触核蛋白毒性的年龄相关增加的遗传调节3）确认实验有效性和对PD的影响，在前述具体目的中获得的每个数据。

DESCRIPTION (provided by applicant): Mutations and environmental factors are known to induce various post-translational alterations in alpha- synuclein, with alterations in alpha-synuclein playing an important role in the onset and progression of Parkinson's disease (PD). In particular, increases in alpha-synuclein insolubility and alpha-synuclein aggregation appear to be central events in the pathogenesis and neurotoxicity observed in PD, as well as experimental models of PD. Aging is a major risk factor for PD and related synucleinopathies, although the underlying genetic basis for age-related alterations to alpha-synuclein is not currently known. Identifying each of the genes which negatively, as well as positively, regulate age-related changes in alpha-synuclein within post-mitotic neurons is likely to lead to a better understanding of the molecular basis for the onset and progression of pathogenesis in PD, as well as potentially lead to the development of novel therapeutics for the treatment of PD. A number of mutations in alpha-synuclein have been identified in recent years, with mutations in alpha-synuclein observed to promote some familial forms of PD. As with aging, it is important to identify the genes which positively and negatively regulate the pathogenesis and, toxicity associated with mutant alpha-synuclein in post-mitotic neurons. The Saccharomyces cerevisiae (S. cerevisiae) model system has been demonstrated to be useful for studying cellular aging, with the stationary phase model of aging contributing to our understanding of post- mitotic cell aging. Additionally, S. cerevisiae have proved useful in studying the biology of many human genes, including mutant human genes associated with disease. The focus of the current proposal is to test the hypothesis that mutagenesis experimentation with the S. cerevisiae stationary phase model of aging aid in developing an understanding of the genetic basis for age-related alterations to wild-type and mutant alpha- synuclein aggregation/insolubility, and age-related increases in wild-type and mutant alpha-synuclein toxicity. Studies will be conducted in cells expressing wild-type and mutant (A53T.A30P) alpha-synuclein. The specific aims are as follows: 1) To utilize mutagenesis to understand the genetic basis for the age-related alterations which occur to alpha-synuclein 2) To utilize mutagenesis to understand the genetic regulation of age-related increases in alpha-synuclein toxicity 3) To confirm the experimental validity, and implications for PD, each of the data obtained in the preceding specific aims.