Embryonic Stem Cell Model of Polyglutamine Disease

多聚谷氨酰胺疾病的胚胎干细胞模型

• 批准号: 6685400
• 负责人: MATTHEW T LORINCZ
• 金额: $ 16.51万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2008-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6685400
• 关键词: Huntington's disease; NMDA receptors; acetylation; brain derived neurotrophic factor; cAMP response element binding protein; embryonic stem cell; genetic transcription; histones; homopeptide; molecular pathology; nerve /myelin protein; neural degeneration; protein biosynthesis; tissue /cell culture

DESCRIPTION (provided by applicant): Huntington's disease (HD) is one of nine fatal neurodegenerative disorders without effective treatment or cure, caused by an expanded CAG/polyglutamine repeat. The roles of the normal and mutant HD gene product, huntingtin remain uncertain. The objective of the proposed Mentored Clinical Scientist Development Award is to explore huntingtin mediated neurodegeneration. We propose to evaluate the roles of transcriptional dysregulation and protein processing in HD by pursuing the following Specific Aims: Specific Aim 1: Define the role of the NMDA receptor subunit NR2B, CBP, histone acetylation, and BDNF in aberrant neurite outgrowth and decreased survival of embryonic stem (ES) cells with expanded CAG repeats. Specific Aim 2: Identify transcripts involved in expanded polyglutamine-mediated transcriptional dysregulation. Specific Aim 3: Characterize the processing of polyglutamine containing proteins in undifferentiated and neuronally differentiated ES cells. We will pursue these Specific Aims utilizing an accurate, easily accessible, murine ES cell model. We find that neuronally differentiated ES cells with expanded CAG repeat domains develop features consistent with polyglutamine-mediated toxicity. Proposed experiments are expected to characterize full-length huntingtin protein processing and identify if the NMDA receptor subunit NR2B, CBP, histone acetylation, or BDNF are mediating neuronal dysfunction that precedes neurodegeneration in HD. Comparison of transcriptional profiles from neuronally differentiated ES cells with and without CAG repeats will be utilized to identify novel factors involved in HD pathogenesis. Once the pathologic mechanisms of mutant huntingtin are understood it will become possible to design rational therapeutics.

描述（由申请人提供）：亨廷顿病（HD）是九种致命的神经退行性疾病之一，无法有效治疗或治愈，由扩展的 CAG/聚谷氨酰胺重复序列引起。正常和突变的 HD 基因产物亨廷顿蛋白的作用仍不确定。拟议的指导临床科学家发展奖的目的是探索亨廷顿蛋白介导的神经变性。我们建议通过追求以下具体目标来评估转录失调和蛋白质加工在 HD 中的作用： 具体目标 1：定义 NMDA 受体亚基 NR2B、CBP、组蛋白乙酰化和 BDNF 在异常神经突生长和存活率降低中的作用具有扩展的 CAG 重复序列的胚胎干 (ES) 细胞。具体目标 2：鉴定参与扩展的多聚谷氨酰胺介导的转录失调的转录本。具体目标 3：表征未分化和神经元分化 ES 细胞中含有聚谷氨酰胺的蛋白质的加工过程。我们将利用准确、易于获取的鼠 ES 细胞模型来实现这些具体目标。我们发现具有扩展的 CAG 重复结构域的神经分化 ES 细胞发展出与多谷氨酰胺介导的毒性一致的特征。拟议的实验预计将表征全长亨廷顿蛋白加工，并确定 NMDA 受体亚基 NR2B、CBP、组蛋白乙酰化或 BDNF 是否介导 HD 神经变性之前的神经元功能障碍。比较具有和不具有 CAG 重复的神经元分化 ES 细胞的转录谱将用于识别 HD 发病机制中涉及的新因素。一旦了解突变型亨廷顿蛋白的病理机制，就有可能设计出合理的治疗方法。