Analysis of Genomic and Proteomic Changes in Glia That Lead To Hydrocephalus

描述（由申请人提供）：本研究项目的目标是确定治疗脑积水的治疗靶点。脑积水（HC）是一种主要的临床疾病，影响美国约0.1%的人口，每年花费超过10亿美元。目前，还没有基于药物的治疗方法来治疗HC。所有HC病例均采用分流术治疗，以降低心室压力。超过30%的新分流器在第一年内失败，超过三分之二的新分流器在十年内失败。此外，2.7%接受分流术的HC患者死于与外科手术相关的并发症。尽管有这些统计数据，但治疗HC的一般方法在50多年来没有显著变化。HC的治疗缺乏进展，在很大程度上是因为HC的病因是多因素的，涉及基因突变和环境损伤。此外，主要负责HC发展的细胞基础和分子途径仍在很大程度上未知。已经开发了许多HC的动物模型;然而，除了HC之外，大多数动物模型还具有多种病理。因此，很难理清HC发展中重要的细胞机制。对人类胎儿的研究，包括不同突变引起的HC，表明室管膜剥脱，随后关闭的导水管的Sylvius发生在大多数情况下的先天性HC。这些和其他研究支持以下假设，即大多数形式的HC中发生的早期事件是心室室管膜细胞的损失或功能障碍以及随后的Sylvius导水管（脑CSF的主要出口通路）的关闭。我们提出了两组实验。首先，我们建议使用一个新开发的小鼠模型HC，以确定发生在室管膜细胞的基因组和蛋白质组的变化，发展之前，明显的HC。这种新的HC模型，称为Ro 1HC，是基于在表达胶质细胞酸性蛋白（GFAP）的细胞中Gi偶联的GPCR的过表达。我们使用四环素诱导的调控系统，使我们能够控制表达GFAP的细胞中的基因表达的时间和水平，使这个模型。重要的是，在这些小鼠中观察到的病理学与在人HC中观察到的病理学非常相似，包括早期室管膜剥脱和随后的Sylvius导水管关闭。其次，我们建议在体外确定相关的HC，可用于筛选能够干扰HC的发展或维持的小分子。 公共卫生相关性：这项拨款提案的目标是确定导致脑积水的关键机制，并开发脑积水的体外相关性，可用于筛选能够治疗这种疾病的小分子。脑积水是一种主要的临床疾病，影响美国约0.1%的人口，每年花费超过10亿美元。目前脑积水的唯一治疗方法是通过脑内植入分流器来缓解脑室内压力。超过30%的新分流术在第一年失败，超过三分之二的分流术在十年内失败; 2.7%接受分流术的脑积水患者死于与外科手术相关的并发症。

DESCRIPTION (provided by applicant):The goal of this research project is to identify therapeutic targets for the treatment of hydrocephalus. Hydrocephalus (HC) is a major clinical disorder affecting ~0.1% of the population in the United States and costing this country more than a billion dollars annually. At this time, there are no drug based therapeutic approaches for treating HC. All cases of HC are treated using shunts to decrease ventricular pressure. More than 30% of all new shunts fail in the first year and more than two thirds of new shunts fail during a ten year period. Further, 2.7 % of HC patients receiving shunts die from complications associated with the surgical procedure. In spite of these statistics, the general approach to treating HC has not changed significantly in over fifty years. The lack of progress in the treatment of HC stems, in large part, from the fact that the causes of HC are multifactorial involving both genetic mutations and environmental insults. Further, the cellular basis and molecular pathways primarily responsible for the development of HC remain largely unknown. A number of animal models of HC have been developed; however, most of these have multiple pathologies in addition to HC. As a result, it has been difficult to sort out cellular mechanisms important in the development of HC. Studies with human fetuses, which include HC arising from different mutations, suggest that ependymal denudation followed by closure of the aqueduct of Sylvius occurs in most cases of congenital HC. These and other studies support the hypothesis that an early event occurring in most forms of HC is the loss or dysfunction of ventricular ependymal cells and consequent closure of the aqueduct of Sylvius, the primary exit pathway for brain CSF. We propose two sets of experiments. First, we propose to use a newly developed mouse model of HC to identify genomic and proteomic changes that occur in ependymal cells prior to the development of overt HC. This new model of HC, referred to as Ro1HC, is based on the over expression of a Gi coupled GPCR in cells expressing glial fibrillary acidic protein (GFAP). We made this model using a tetracycline inducible regulatory system that enables us to control the timing and level of gene expression in cells expressing GFAP. Importantly, pathology observed in these mice closely resembles that observed in human HC, including the early ependymal denudation and subsequent closure of the aqueduct of Sylvius. Second, we propose to identify in vitro correlates of HC that can be used to screen for small molecules capable of interfering with the development or maintenance of HC. PUBLIC HEALTH RELEVANCE: The goal of this grant proposal is to identify the key mechanisms that lead to hydrocephalus as well as develop an in vitro correlate of hydrocephalus that can be used to screen for small molecules capable of treating this disease. Hydrocephalus is a major clinical disorder affecting ~0.1% of the population in the United States and costing this country more than a billion dollars annually. The only current treatment of hydrocephalus is based on surgically-implanting shunts designed to relieve intraventricular pressure. More than 30% of all new shunts fail in the first year and more than two thirds of shunts fail within ten years; 2.7% of hydrocephalic patients receiving shunts die from complications associated with the surgical procedure.