Role of Proteasome Inhibition in the Pathogenesis of CADASIL

蛋白酶体抑制在 CADASIL 发病机制中的作用

基本信息

批准号：
7600370
负责人：
Michael M Wang
金额：
$ 13.34万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2011-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7600370
关键词：
26S proteasome Affect Alzheimer&apos s Disease Amyotrophic Lateral Sclerosis Arteries Blood Vessels CADASIL Cell Culture Techniques Cell Death Cell Density Cell Line Cell Separation Cells Cessation of life Chronic Complex Cysteine Data Defect Dementia Disease Family Functional disorder Genes Genetic Predisposition to Disease Goals Hereditary Disease Human Huntington Disease Injury Intercellular Junctions Language Learning Light Link Modeling Molecular Mutation Neurodegenerative Disorders Parkinson Disease Pathogenesis Pathway interactions Play Point Mutation Prevention strategy Proteasome Inhibition Proteins Research Role Series Smooth Muscle Smooth Muscle Myocytes Step Tests Stroke System Testing Tissues Toxic effect Transgenic Animals Ubiquitin Up-Regulation Vascular Dementia Vascular Diseases Vascular Smooth Muscle Work cytotoxicity design gain of function in vivo interest mouse model multicatalytic endopeptidase complex mutant overexpression premature protein degradation protein expression protein protein interaction research study

项目摘要

DESCRIPTION (provided by applicant): Stroke and vascular dementia are prevalent diseases which are both caused by multiple factors, including genetic susceptibility. In large part, the genetics of these disorders is complex. However, in families with CADASIL (Cerebral Autosomal Dominant Arteriopathy with Subcortical Infarcts and Leukoencephalopathy), premature stroke and vascular dementia are caused by defects in the Notch3 gene. Our broad goal is to identify the molecular pathways governing the pathogenesis of CADASIL, which will help shed light on more common forms of stroke and dementia. The central hypothesis is that mutant NotchS, expressed in arterial smooth muscle, triggers a series of cellular changes that result in inhibition of the proteasome. The proteasome is essential for the degradation of proteins via the ubiquitin proteasome system and has been implicated in several neurodegenerative disorders. We propose that inhibition of the proteasome in vascular smooth muscle causes cell toxicity and is linked to the breakdown of cell-cell contacts and accumulation of Notch3 protein, two pathological features of CADASIL. In support of this concept, we present preliminary data showing that Notch3 specifically interacts with a subunit of the 26S proteasome. In addition, we demonstrate evidence that proteasome inhibition is increased in cells expressing mutant Notch3. Finally, proteasome inhibition in cultured smooth muscle cells causes cell-cell separation and cell death, two features seen in CADASIL tissues. In Specific Aim 1, we will characterize whether mutant Notch3 directly inhibits proteasomes and whether mutant Notch3 over-expression leads to proteasome inhibition in cell culture and in arteries of transgenic animals. In Aim 2, we will determine whether proteasome inhibition leads to cell-cell contact disruption and Notch3 protein accumulation in culture and in vivo. Finally, in Aim 3, we will investigate whether cell death due to proteasome inhibition is exacerbated by Notch3 mutations and by cell-cell contact disruption. These experiments will support a CADASIL pathway involving proteasomal inhibition and may have implications for the pathogenesis of other vascular and proteasome-linked diseases. In lay language, this application is designed to investigate the molecules involved in a disease, CADASIL that causes stroke and dementia. An understanding of CADASIL may provide clues to designing better treatments and prevention strategies for stroke and dementia. What is learned from this study may also provide information that will be useful for the treatment of other vascular diseases.

描述（由申请人提供）：中风和血管性痴呆是普遍的疾病，它们都是由多种因素引起的，包括遗传敏感性。在很大程度上，这些疾病的遗传学很复杂。然而，在卡达西尔（脑常染色体显性动脉炎和皮质下梗塞和白细胞脑病）的家庭中，早产和血管性痴呆是由Notch3基因缺陷引起的。我们的广泛目标是确定控制卡达西发病机理的分子途径，这将有助于阐明中风和痴呆症的更常见形式。中心假设是在动脉平滑肌肉中表达的突变档触发一系列细胞变化，从而抑制蛋白酶体。蛋白酶体对于通过泛素蛋白酶体系统降解蛋白质至关重要，并且与几种神经退行性疾病有关。我们建议抑制血管平滑肌中蛋白酶体会引起细胞毒性，并与细胞 - 细胞接触的分解和Notch3蛋白的积累（Cadasil的两个病理特征）的积累有关。为了支持这个概念，我们提供了初步数据，表明Notch3专门与26S蛋白酶体的亚基相互作用。此外，我们证明了表达突变型notch3细胞中蛋白酶体抑制增加的证据。最后，培养的平滑肌细胞中的蛋白酶体抑制会导致细胞间的分离和细胞死亡，这两个特征在卡达西尔组织中看到。在特定的目标1中，我们将表征突变体Notch3是否直接抑制蛋白酶体以及突变Notch3过表达是否会导致细胞培养和转基因动物动脉的蛋白酶体抑制。在AIM 2中，我们将确定蛋白酶体抑制是否会导致细胞 - 细胞接触破坏和Notch3蛋白在培养和体内的积累。最后，在AIM 3中，我们将研究Notch3突变和细胞 - 细胞接触破坏会导致蛋白酶体抑制作用引起的细胞死亡。这些实验将支持涉及蛋白酶体抑制的卡达西尔途径，并可能对其他血管和蛋白酶体连接疾病的发病机理有影响。在外行语言中，该应用旨在调查引起中风和痴呆的疾病的分子。对卡达西尔的了解可能会为设计更好的治疗方法和预防策略提供有关中风和痴呆症的线索。从这项研究中学到的内容还可能提供对治疗其他血管疾病的信息。