Identification of pathogenic mechanisms important in multiple system atrophy

 描述（由申请人提供）：有许多例子表明，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”，通过它可以理解这种疾病的机制。这项拨款提案的重点是多系统萎缩（MSA），一种罕见的，但严重的神经退行性突触核蛋白病与成人发病的特点是自主神经功能障碍与帕金森综合征和/或小脑症状的调查。在MSA中特别感兴趣的是由纤维状α-突触核蛋白（α-SYN）蛋白形成的少突胶质细胞和神经元胞质内含物（GCI和NCI），尽管驱动这些积累的致病机制尚不清楚。目前，没有预防或改变MSA的疾病治疗方法，并且迫切需要研究以确定可以靶向治疗的途径。很少有研究已经做了了解这种疾病的病因，特别是在现代基因组测序领域，即使这样的方法已经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相关的罕见遗传变异。我们将对MSA患者、父母和一名未受影响的兄弟姐妹的四胞胎进行基因组测序。鉴定的遗传变异体将在 通过使用现有细胞系的CRISPR-Cas9工程化进行体外研究。我们的第二个目标是通过将激光捕获显微切割和下一代RNA-Seq结合在神经病理证实的MSA脑和匹配对照中，鉴定对MSA a-SYN积累重要的致病机制。这两个目标都旨在确定潜在的疾病机制和关键靶点，可作为未来潜在的治疗选择。我们的发现可能会进一步阐明更常见的突触核蛋白病如帕金森病和路易体痴呆症的发病机制。

 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.