PATHOLOG-OMICS - ESSENTIAL TREMOR IN THE BROADER CONTEXT OF

在过去的5 - 8年里，我们已经确定了一组基本的形态学变化， 震颤（ET）小脑，主要集中在/周围浦肯野细胞（PC）。发现 ET相关病理学的研究引起了极大的兴趣，但也提出了一些棘手的问题。 在原发性小脑神经退行性病变中也观察到了其中的几种病理。 疾病，如脊髓小脑共济失调（SCA）和多系统萎缩（MSA），但 这些变化发生的程度尚未正式研究或与 ET.有趣的是，小脑现在越来越多地与震颤的产生有关， 其他疾病，如帕金森病（PD）和肌张力障碍，但他们的小脑病理是 目前未开发。因此，存在很大的形态学数据缺口。在更初级的层面上， 我们最近进行了激光捕获显微切割（LCM），专门针对PC，从而 有助于精确评估ET中细胞特异性分子变化。我们得到了一个高度 新的差异基因表达谱通过RNA的直接测序（RNA-seq）分离自 ET的PC与对照大脑。我们鉴定了47个差异表达的转录本， 调节神经元功能的蛋白质。然而，一组平行的LCM-RNA-seq研究， 探索PD、肌张力障碍、MSA和SCA中PC的分子生物学， 执行。这代表了第二个大的分子数据缺口。这份为期五年的提案， 使用来自ET患者以及一系列其他疾病患者的死后组织， 小脑疾病，有两个目的。具体目标1：开展详细的 小脑，比较ET患者小脑的形态学变化， 原发性小脑退行性疾病（SCA和MSA）患者以及 伴有震颤和小脑受累（PD和肌张力障碍）的神经系统疾病患者。 我们将收集160个大脑的初始发现样本（50个ET，25个SCA，15个MSA，30个PD，15个MS）。 肌张力障碍，25个对照）以及160个大脑的重复样品，评估病理学 改变了几个小脑隔间。量化的聚类分析 变量将被用来确定是否有一个可定义的“ET集群”以及可定义的 其他四种疾病的集群。具体目标2：探索分子生物学 以小脑受累为特征的神经退行性疾病中的PC和/或 震颤使用新的LCM-RNA-seq方法，我们将确定分子表达是否 这些疾病中PC的变化是相同或不同的。对于这些新的分子 在这些研究中，我们建议使用60个脑（ET、SCA、MSA、PD、肌张力障碍、对照各10个）。

Over the past 5 - 8 years, we have identified a cluster of morphological changes in the essential tremor (ET) cerebellum, predominantly centered in/around the Purkinje cell (PC). The discovery of ET-related pathology has generated great interest but it has also raised difficult questions. Several of these pathologies have also been observed in primary cerebellar neurodegenerative diseases such as spinocerebellar ataxias (SCAs) and multiple system atrophy (MSA), but the degree to which these changes occur has not been formally studied or compared with that in ET. Interestingly, the cerebellum is now increasingly being implicated in tremor generation in other diseases such as Parkinson's disease (PD) and dystonia, yet their cerebellar pathology is presently unexplored. Hence, there is a large morphologic data gap. On a more primary level, we recently performed laser-capture microdissection (LCM) to specifically target PCs, thereby facilitating a precise evaluation of cell-specific molecular changes in ET. We obtained a highly novel differential gene expression profile by direct sequencing of RNA (RNA-seq) isolated from PCs of ET vs. control brains. We identified 47 differentially expressed transcripts, which code for proteins that regulate neuronal function. However, a parallel set of LCM-RNA-seq studies, exploring the molecular biology of PCs in PD, dystonia, MSA and SCA, have yet to be performed. This represents a second, large molecular, data gap. This five-year proposal, which uses postmortem tissue from patients with ET well as from patients with a range of other cerebellar disorders, has two aims. Specific Aim 1: To undertake detailed postmortem studies of the cerebellum, comparing morphological changes in the cerebellum of ET patients to those of patients with primary cerebellar degenerative diseases (SCA and MSA) as well as those of patients with neurological diseases with tremor and cerebellar involvement (PD and dystonia). We will assemble an initial discovery sample of 160 brains (50 ET, 25 SCA, 15 MSA, 30 PD, 15 dystonia, 25 controls) as well as a replicate sample of 160 brains, assessing pathological changes across several cerebellar compartments. Hierarchical cluster analysis of quantified variables will be used to determine whether there is a definable “ET cluster” as well as definable clusters for each of these other four diseases. Specific Aim 2: To explore the molecular biology of PCs across neurodegenerative diseases characterized by cerebellar involvement and/or tremor. Using a novel LCM-RNA-seq approach, we will determine whether molecular expression changes in PCs are the same or differ across these diseases. For these novel molecular studies, we propose to use 60 brains (10 each of ET, SCA, MSA, PD, dystonia, controls).