Spinocerebellar Ataxia Type 10

描述（改编自申请人摘要）：脊髓小脑共济失调10型 (SCA 10）是一种以小脑共济失调为特征的常染色体显性疾病， 癫痫发作和预期。SCA 10突变是一个大规模（4-22 kb）扩增 在E46的内含子9中的ATTCT五核苷酸重复，E46是一个未知的新基因， 功能，在染色体22q13.3。该项目的长期目标是 阐明SCA 1 0的分子机制，制定合理的治疗方案 基于这种渐进的致病机制的策略 使人衰弱的疾病本申请将调查的机制， 扩展的ATTCT重复序列导致疾病表型。的假设 拟议的项目是，ATTCT重复的扩大导致疾病， 通过重复序列大小依赖性机制改变E46转录物。SCA1 0 是一个新认识的显性遗传性共济失调，其 流行病学、临床和遗传特征尚未完全 表征了因此，进一步表征基因型、表型和 SCA 10患者的基因型-表型相关性需要作为起点 该项目的目标，并指定为具体目标1。完成这一 具体目标应该加强我们的假设，即致病因素 机制取决于重复的大小。具体目标2将检查E46 在可用的组织中寻找数量变化， SCA 10中加工的转录物和转录物的异常定位。 具体目标3将通过分析E46蛋白的功能来研究E46蛋白的功能。 通过蛋白质印迹法测定该蛋白质的量和同种型， 通过免疫细胞化学进行蛋白质定位，分离相互作用的蛋白质 与正常的和异常的E46蛋白产物（如果检测到的话）， 酵母双杂交和免疫共沉淀技术， 用于细胞存活、增殖和增殖的E46缺陷型细胞培养模型 分化研究。从具体目标2和3获得的数据将 与扩大的重复规模相比，如果适用的话。完成这些 具体目标将提供重要的见解，了解致病性 SCA 10中这种新型五核苷酸扩增的结果。

DESCRIPTION (Adapted from applicant's abstract): Spinocerebellar ataxia type 10 (SCA 10) is an autosomal dominant disease characterized by cerebellar ataxia, seizure and anticipation. The SCA1O mutation is a massive (4-22 kb) expansion of the ATTCT pentanucleotide repeat in intron 9 of E46, a novel gene of unknown function, on chromosome 22q13.3. The long-term objective of this project is to elucidate the molecular mechanisms of SCA1 0, and to develop rational treatment strategies based on the disease-causing mechanism for this progressive debilitating disease. This application will investigate the mechanism by which the expanded ATTCT repeat leads to the disease phenotype. The hypothesis of the proposed project is that an expansion of the ATTCT repeat causes the disease by altering the E46 transcript through a repeat size-dependent mechanism. SCA1 0 is one of the newly recognized dominantly inherited ataxias, and its epidemiological, clinical and genetic features have not been fully characterized. Thus, further characterization of the genotype, phenotype, and genotype-phenotype correlation in SCA10 patients needs to be the starting point of this project, and is designated as Specific Aim 1. Accomplishing this Specific Aim should strengthen our hypothesis that the disease-causing mechanism is dependent on the repeat size. Specific Aim 2 will examine the E46 transcripts in available tissues to look for quantitative changes, aberrantly processed transcripts and abnormal localization of transcripts in SCA10. Specific Aim 3 will investigate E46 protein functions by analyzing the quantities and isoforms of this protein by western blot, determining the protein localization by immuno-cytochemistry, isolating proteins that interact with the normal and, if detected, aberrant E46 protein products by yeast-two-hybrid and co-immunoprecipitation techniques, and developing E46-deficient cell culture models for cell survival, proliferation and differentiation studies. Data obtained from Specific Aims 2 and 3 will be compared with the expanded repeat size, when applicable. Accomplishing these Specific Aims will provide important insights into understanding the pathogenic consequences of this novel pentanucleotide expansion in SCA10.