Genetic basis for alpha-synuclein toxicity

描述(申请人提供)：已知突变和环境因素可导致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氏病(PD)的发生和发展中起着重要作用。特别是，α-突触核蛋白不溶性和α-突触核蛋白聚集性的增加似乎是帕金森病发病机制和神经毒性观察到的中心事件，以及帕金森病的实验模型。衰老是帕金森病和相关联核病的主要危险因素，尽管与年龄相关的α-联核蛋白改变的潜在遗传基础目前尚不清楚。识别每一个负向和正向调节有丝分裂后神经元中与年龄相关的α-突触核蛋白变化的基因，可能有助于更好地理解帕金森病发病和发展的分子基础，并有可能导致治疗帕金森病的新疗法的发展。近年来，已经发现了一些α-突触核蛋白的突变，观察到α-突触核蛋白的突变会促进某些家族性帕金森病的发生。与衰老一样，重要的是确定与有丝分裂后神经元中突变的α-突触核蛋白相关的正向和负向调控发病机制和毒性的基因。酿酒酵母(S.cerevisiae)模型系统已被证明是研究细胞衰老的有用模型，衰老的稳定期模型有助于我们理解有丝分裂后的细胞衰老。此外，酿酒酵母已被证明在研究许多人类基因的生物学上很有用，包括与疾病相关的突变人类基因。目前建议的重点是检验这样一种假设，即用酿酒酵母衰老的固定相模型进行突变实验有助于理解与年龄相关的野生型和突变型α-突触核蛋白聚集/不溶解变化的遗传基础，以及与年龄相关的野生型和突变型α-突触核蛋白毒性的增加。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。其具体目的如下：1)利用诱变来了解α-突触核蛋白发生年龄相关性改变的遗传基础2)利用诱变来了解与年龄相关的α-突触核蛋白毒性增加的遗传规律3)确认实验的有效性，以及对帕金森病的影响，每个数据都在前面的特定目的中获得。

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.