Alternative Splicing of ApoER2 in Alzheimer's Disease

阿尔茨海默病中 ApoER2 的选择性剪接

• 批准号: 10396990
• 负责人: Christina Marie Gallo
• 金额: $ 1.89万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-07 至 2022-04-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10396990
• 关键词: Address; Affect; Age; Aging; Alleles; Alternative Splicing; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease risk; Amyloid beta-Protein; Antisense Oligonucleotides; Apolipoprotein E; Arginine; Autopsy; Binding; Binding Sites; Biochemical; Bioinformatics; Biological Assay; Biology; Brain; Brain region; Cerebellum; Cognition; Development; Disease; Disease Progression; Disease susceptibility; Electrophoretic Mobility Shift Assay; Equilibrium; Event; Exons; Fellowship; Genes; Goals; Hippocampus (Brain); Human; Immunoprecipitation; Impaired cognition; In Vitro; Individual; LDL-Receptor Related Protein 1; Late Onset Alzheimer Disease; Learning; Link; Lipids; Long-Term Potentiation; Maps; Mass Spectrum Analysis; Mediating; Memory; Modeling; Molecular; Mus; Nature; Nerve Degeneration; Neurons; Parietal Lobe; Pathogenesis; Pathology; Pathway interactions; Phosphorylation; Phosphotransferases; Physiological; Post-Translational Protein Processing; Post-Translational Regulation; Prefrontal Cortex; Process; Protein Isoforms; Proteins; Publishing; RNA; RNA Splicing; Regulation; Reporter; Research; Risk; Risk Factors; Role; Serine; Signal Transduction; Signaling Molecule; Site; Spliced Genes; Synapses; Synaptic plasticity; Techniques; Therapeutic Human Experimentation; Training; Transcript; Variant; Work; apolipoprotein E receptor 2; brain tissue; experience; extracellular; insight; interest; mRNA Precursor; mind control; mutant; neuropathology; novel; receptor; response; single molecule; support network; transcriptome sequencing; translational potential

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.

项目摘要：年龄和载脂蛋白 E (apoE) 的 ɛ4 等位基因是两个最大的风险因素 发展为晚发性阿尔茨海默病 (LOAD)。由于 LOAD 没有已知的治疗方法或原因， 了解已知风险因素的机制对于推动治疗研究至关重要 向前。 ApoE 通过与 apoER2 等同源受体结合来介导其在大脑中的生理作用， 存在于多种受精确调控的剪接亚型或变体中。最近的研究发现了一个 衰老、神经退行性变和选择性剪接之间的新联系。有趣的是，选择性剪接 apoE 受体 apoER2 在阿尔茨海默病 (AD) 中发生改变。因此，衰老过程中异常的选择性剪接 神经退行性变可能通过 apoER2 改变关键 AD 危险因素 APOE 的相互作用组，影响 APOE 生理功能并突出了研究 APOE 作用的新科学范式 APOE 处于负载状态。该提案的总体目标是研究 apoER2 的剪接是否发生改变 AD 并发现 apoER2 剪接事件的修饰因子。中心假设是 apoER2 亚型 由于剪接因子翻译后调节的改变，AD 中的平衡受到干扰，这与 整个大脑的病理发展。为了研究这一假设，提出了两个具体目标。瞄准 1，apoER2 的亚型分布将在 AD Braak 阶段和脑区域中确定，与 使用单分子长读长测序和 qPCR 的健康个体。在目标 2 中，对特定的监管 剪接因子引起的 apoER2 剪接事件将使用多种体外技术进行探测，包括 RNA- 免疫沉淀、剪接报告基因测定和凝胶位移测定。该提案的完成将获得高度 意义重大，提供了一种新颖的科学模型，通过该模型可以查看 AD 中赋予的 APOE 风险：改变 同源受体 apoER2 的剪接调节。该提案结合了生物信息学、突触学 将生物学和 RNA 生物学纳入一个综合培训计划，为申请人提供全面和丰富的培训 奖学金培训经历。这是通过广泛的支持赞助商和 承诺指导申请人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。