Alternative Splicing of ApoER2 in Alzheimer's Disease

项目概述：年龄和载脂蛋白E（apoE）的等位基因是两个最大的危险因素， 晚发性阿尔茨海默病（LOAD）由于没有已知的治疗方法或原因的负载， 了解已知危险因素的潜在机制对于推动治疗研究至关重要。 性新ApoE通过与apoER 2等同源受体结合来介导其在脑中的生理作用， 存在于多个精确调控的剪接异构体或变体中。最近的研究发现， 衰老、神经退化和可变剪接之间的新联系。有趣的是， apoE受体apoER 2在阿尔茨海默病（AD）中改变。因此，在衰老过程中， 神经退行性变可能通过apoER 2改变AD关键危险因子APOE的相互作用， APOE的生理功能，并突出了一个新的科学范式，通过它来调查的作用， APOE在加载中。这项研究的总体目标是研究apoER 2的剪接是否被改变， 并揭示apoER 2剪接事件的修饰剂。核心假设是apoER 2亚型 由于剪接因子的翻译后调节改变，AD中的平衡被扰乱， 大脑的病理发展。为了研究这一假设，提出了两个具体的目标。在Aim中 1中，将确定apoER 2的亚型分布在AD Braak分期和脑区域中，与 使用单分子长读段测序和qPCR对健康个体进行检测。在目标2中， 将使用多种体外技术，包括RNA-聚合酶链反应（RT-PCR），检测剪接因子引起的apoER 2剪接事件。 免疫沉淀、剪接报告基因测定和凝胶迁移测定。完成这一提案将是高度 重要的是，提供了一个新的科学模型，通过它来查看赋予AD的APOE风险：改变 同源受体apoER 2的剪接调节。该提案将生物信息学、突触学 生物学和RNA生物学为一体的综合培训计划，为申请人提供全面而丰富的 研究金培训经验。这是通过一个广泛的支持赞助商网络来实现的， 合作者谁已承诺指导申请人通过拟议的研究，并促进她 成为一名独立的神经科学家。

PROJECT SUMMARY: Age and the ɛ4 allele of apolipoprotein E (apoE) are the two greatest risk factors for developing Late Onset Alzheimer’s Disease (LOAD). As there is no known cure for or cause of LOAD, understanding the mechanism underlying known risk factors is paramount for moving therapeutic research forward. ApoE mediates its physiological effects in the brain by binding to cognate receptors like apoER2, which is found in multiple splice isoforms or variants that are precisely regulated. Recent studies have discovered a novel link between aging, neurodegeneration and alternative splicing. Interestingly, alternative splicing of the apoE receptor apoER2 is altered in Alzheimer’s disease (AD). Therefore, abnormal alternative splicing in aging and neurodegeneration may alter the interactome of the key AD risk factor APOE through apoER2, influencing APOE physiological function and highlighting a new scientific paradigm through which to investigate the role of APOE in LOAD. The overall goal of this proposal is to investigate whether splicing of apoER2 is altered in AD and to uncover modifiers of apoER2 splicing events. The central hypothesis is that apoER2 isoform balance is perturbed in AD due to altered posttranslational regulation of splicing factors, correlating with pathology development across the brain. To investigate this hypothesis, two specific aims are proposed. In Aim 1, the isoform distribution of apoER2 will be determined across AD Braak stage and brain region compared to healthy individuals using single molecule long read sequencing and qPCR. In Aim 2, regulation of a specific apoER2 splicing event by a splicing factor will be probed using a variety of in vitro techniques including RNA- Immunoprecipitations, splicing reporter assays and gel shift assays. Completion of this proposal will be highly significant, providing a novel scientific model through which to view conferred APOE risk in AD: altered splicing regulation of the cognate receptor apoER2. This proposal incorporates bioinformatics, synaptic biology and RNA biology into one integrated training plan providing the applicant a well-rounded and rich fellowship training experience. This is accomplished by an expansive network of supporting sponsors and collaborators who have pledged to guide the applicant through the proposed research and facilitate her development into an independent research neuroscientist.