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ɛ4载脂蛋白E（APOE）的等位基因是两个最大的风险因素 发展阿尔茨海默氏病晚期（负载）。由于没有已知的治疗或负载原因，所以 了解危险因素的基本机制对于移动治疗研究至关重要 向前。 ApoE通过与apoer2（如Apoer2）结合的同源受体来介导其在大脑中的身体作用， 在精确调节的多种剪接同工型或变体中发现。最近的研究发现了 衰老，神经变性和替代剪接之间的新型联系。有趣的是，替代剪接 Apoe受体apoer2在阿尔茨海默氏病（AD）中发生了变化。因此，衰老中的异常替代剪接 神经变性可能会通过Apoer2（影响者）改变关键AD风险因素APOE的相互作用 APOE的身体功能并突出了一种新的科学范式，以调查该范式 负载中的apoe。该提案的总体目标是调查apoer2的剪接是否改变 在AD中并发现ApoER2剪接事件的修饰符。中心假设是Apoer2同工型 由于剪接因子的翻译后调节改变，平衡在AD中受到干扰，与之相关 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。目标 1，与APOER2的同工型分布将在AD Braak阶段和大脑区域确定 使用单分子长读取测序和qPCR的健康个体。在AIM 2中，对特定的调节 通过多种体外技术（包括RNA- 免疫沉淀，剪接记者测定法和凝胶转移测定法。该提议的完成将是高度的 重要的是，提供了一种新颖的科学模型，通过该模型可以在AD中查看会议的APOE风险： 同源受体apoer2的剪接调节。该提案结合了生物信息学，突触 生物学和RNA生物学成为一个综合培训计划，为申请人提供了全面且丰富的 奖学金培训经验。这是通过支持赞助商和的广泛网络来完成的 通过拟议的研究指导适用并促进她的合作者 发展成独立的研究神经科学家。