Protein Trafficking in Neurodegenerative Diseases

神经退行性疾病中的蛋白质贩运

• 批准号: 7454328
• 负责人: Ai Yamamoto
• 金额: $ 35.3万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-20 至 2009-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7454328
• 关键词: Affect; Animals; Area; Biochemical Genetics; Biological Assay; Cell Line; Cells; Chemicals; Clear Cell; Degradation Pathway; Disease; Gene Expression; Gene Silencing; Genes; Genetic; Genetic Techniques; Grant; Green Fluorescent Proteins; Huntington Disease; Lead; Lysosomes; Mediating; Modeling; Mutation; Neurodegenerative Disorders; Neurotransmitters; Pathway interactions; Proteins; Recovery; Research Personnel; Running; Small Interfering RNA; Source; Specificity; Sum; Symptoms; Testing; Tissues; Variant; base; cell type; genetic profiling; human Huntingtin protein; insight; intracellular protein transport; mutant; polyglutamine; programs; protein aggregation; protein degradation; protein transport; stable cell line; trafficking; trait

DESCRIPTION (provided by applicant): Accumulation and aggregation of mutant proteins are common traits across different neurodegenerative disorders, such as the polyglutamine expansion disorder Huntington's disease (HD). A recently emerging theme is that if mutant protein accumulation is eliminated, symptomatic progression not only halts but also recovers. For example, in an inducible model of Huntington's disease, loss of mutant protein accumulation in symptomatic animals led to complete reversion of the disease-like symptoms. Therefore, if we can accelerate the clearance of a disease-causing mutant protein, there exists the tantalizing possibility of recovery from disease. But how do cells clear these mutant proteins? And how does clearance of the mutant proteins lead to recovery from symptoms? To gain insight into these questions we have run Affymetrix gene arrays on stably transfected cell lines that carry mutant huntingtin protein. The comparison of the different genetic profiles revealed surprisingly robust changes in pathways indicating lysosome-mediated degradation and vesicular trafficking. These two areas are little explored in Huntington's disease and polyglutamine diseases in general, and is thus a rich source of questions. In this proposal we will therefore systematically test the following hypotheses: 1) Lysosome-mediated degradation has a significant impact on the degradation of mutant huntingtin proteins; and 2) Aggregation leads to reversible deficits in vesicular trafficking. Using a combination of biochemical and genetic techniques we also propose to identify regulators of protein aggregation and clearance using a functional cell-based assay: a stable cell line that conditionally expresses mutant proteins fused to variants ol GFP. In sum, during this grant period we will reveal targets that directly alter the level of mutant proteins in a cell, elucidate the basic degradation pathways crucial for handling these difficult proteins, and examine how deficits in protein degradation alters vesicular trafficking in the cell.

描述（由申请人提供）：突变蛋白的积累和聚集是不同神经退行性疾病的常见特征，例如聚谷氨酰胺扩张障碍亨廷顿氏病（HD）。最近一个新兴的主题是，如果消除了突变蛋白的积累，则有症状的进展不仅停止，而且会恢复。例如，在亨廷顿氏病的诱导模型中，有症状动物中突变​​蛋白积累的丧失导致疾病样症状的完全恢复。因此，如果我们可以加速致病的突变蛋白的清除，则存在诱人的疾病恢复的可能性。 但是细胞如何清除这些突变蛋白？突变蛋白的清除如何导致从症状中恢复？为了深入了解这些问题，我们已经在携带突变蛋白蛋白的稳定转染细胞系上运行Affymetrix基因阵列。不同遗传特征的比较表明，途径的稳健变化表明溶酶体介导的降解和囊泡运输。这两个领域在亨廷顿氏病和多谷氨酰胺疾病中几乎没有探索，因此是广泛的问题根源。因此，在此提案中，我们将系统地检验以下假设：1）溶酶体介导的降解对突变型亨廷汀蛋白的降解具有重大影响； 2）聚集会导致囊泡贩运的可逆缺陷。使用生化技术和遗传技术的组合，我们还建议使用基于功能细胞的测定法鉴定蛋白质聚集和清除的调节剂：稳定的细胞系，该稳定的细胞系有条件地表达与变体OL GFP融合的突变蛋白。总而言之，在此赠款期间，我们将揭示直接改变细胞中突变蛋白水平的靶标，阐明对处理这些困难蛋白至关重要的基本降解途径，并研究蛋白质降解的缺陷如何改变细胞中的囊泡运输。