Identification of pathogenic mechanisms important in multiple system atrophy

 描述(申请人提供)：有许多例子表明，对罕见疾病的研究从根本上改变了我们对具有相似表型特征的常见疾病的理解。这在很大程度上是正确的，因为这种罕见的疾病代表了一个更清晰的“透镜”，通过它来了解这种疾病的机制。这项拨款建议重点研究多系统萎缩(MSA)，这是一种罕见但严重的神经退行性突触核病，以帕金森氏症和/或小脑症状的自主神经功能障碍为特征。对MSA特别感兴趣的是由纤维状α-突触核蛋白(a-SYN)蛋白形成的少突胶质细胞和神经元胞浆内含物(GCIS和NCIS)，尽管这些聚集的致病机制尚不清楚。目前，还没有针对MSA的疾病预防或修改治疗方法，迫切需要研究以确定可以作为治疗目标的途径。很少有人研究这种疾病的病因，特别是在现代基因组测序领域，尽管这种方法彻底改变了罕见的儿童疾病的调查方式。我们的第一个目标是使用基于家族的基因组学来发现与MSA相关的罕见基因变异。我们将对MSA患者、父母和一名未受影响的兄弟姐妹进行基因组测序。已识别的遗传变异将在 利用CRISPR-Cas9工程对现有的细胞系进行体外培养。我们的第二个目标是通过结合激光捕获显微解剖和下一代RNA-Seq在神经路径确认的MSA大脑和匹配的对照组中识别MSA a-SYN积累的重要致病机制。这两个目标都是为了确定潜在的疾病机制和关键靶点，这些目标可以被用作未来潜在的治疗选择。我们的发现可能会为帕金森氏病和路易体痴呆等更常见的联体核病的发病机制提供更多的线索。

 DESCRIPTION (provided by applicant): There are many examples where the study of rare disease has fundamentally changed our understanding of common diseases with similar phenotypic characteristics. This is true to a large extent because the rare disease represents a much clearer "lens" by which to understand the mechanisms of the disorder. This grant proposal focuses on the investigation of multiple system atrophy (MSA), a rare but severe neurodegenerative synucleinopathy with adult onset characterized by autonomic dysfunction with parkinsonism and/or cerebellar symptoms. Of specific interest in MSA are the oligodendroglial and neuronal cytoplasmic inclusions (GCIs and NCIs) formed by fibrillar alpha-synuclein (a-SYN) proteins although the pathogenic mechanisms driving these accumulations are unclear. Currently, there are no disease preventing or modifying treatments for MSA, and a crucial need for research to identify pathways that can be targeted for therapy. Little research has been done to understand the etiology of this disease especially in the area of modern genomic sequencing even though such an approach has revolutionized the way rare childhood disorders are investigated. Our first goal is to use family-based genomics to uncover rare genetic variants associated with MSA. We will perform genomic sequencing on quads of MSA patients, parents, and one unaffected sibling. Identified genetic variants will be characterized in vitro by using CRISPR-Cas9 engineering of existing cell-lines. Our second goal is to identify the pathogenic mechanisms important to MSA a-SYN accumulation, by combining laser capture microdissection and next generation RNA-Seq in neuropath-confirmed MSA brains and matched controls. Both of these aims are designed to identify underlying disease mechanisms and key targets that could be exploited as potential future therapeutic options. Our findings may shed additional light on the mechanisms of disease in more common synucleinopathies like Parkinson's disease and Lewy Body Dementia.