Alternative Splicing of ApoER2 in Alzheimer's Disease

项目摘要：年龄和载脂蛋白E的ɛ4等位基因是两个最大的风险因素 发展迟发性阿尔茨海默病(LOAD)。由于目前尚无已知的治疗方法或负荷原因， 了解已知危险因素背后的机制对于推动治疗研究至关重要 往前走。载脂蛋白E通过与ApoER2等同源受体结合来调节其在大脑中的生理效应。 在多种剪接异构体或变异体中被发现，这些剪接异构体或变体受到精确的调控。最近的研究发现， 衰老、神经退化和选择性剪接之间的新联系。有趣的是， 载脂蛋白E受体ApoER2在阿尔茨海默病(AD)中发生改变。因此，衰老过程中的异常选择性剪接 神经退行性变可能通过ApoER2改变AD关键风险因子ApoE的相互作用，影响 APOE的生理功能，并突出了一种新的科学范式，通过它来研究 APOE已装入。这项提案的总体目标是调查ApoER2的剪接是否发生了变化 在AD中，并发现ApoER2剪接事件的修饰。中心假说是ApoER2亚型 由于剪接因子翻译后调节的改变，AD的平衡被扰乱，与 整个大脑的病理发展。为了研究这一假说，我们提出了两个具体目标。在AIM 1，将确定ApoER2在AD Braak分期和脑区的异构体分布 健康个体采用单分子长阅读测序和qPCR。在目标2中，对一种特定的 由剪接因子引起的ApoER2剪接事件将使用各种体外技术进行探测，包括RNA- 免疫沉淀、剪接报告分析和凝胶移位分析。这项提案的完成将极大地 意义重大，提供了一种新的科学模型，通过该模型可以查看AD的APOE风险：已更改 同源受体ApoER2的剪接调控。这项提议结合了生物信息学、突触 生物学和RNA生物学整合为一体的培养计划，为申请者提供全面而丰富的 团契培训经验。这是通过支持赞助商的广泛网络和 承诺指导申请者完成拟议研究并为其提供便利的合作者 发展成为一名独立研究的神经科学家。

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.