Multimodal Characterization of the Role of Circular RNAs in Alzheimer's Disease

环状 RNA 在阿尔茨海默病中作用的多模式表征

• 批准号: 10446362
• 负责人: Carlos Cruchaga
• 金额: $ 226.52万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10446362
• 关键词: Affect; Aging; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease pathology; Alzheimer' s disease patient; Alzheimer' s disease risk; Alzheimer’s disease biomarker; Amyloid beta-Protein; Autopsy; Back; Behavior; Behavioral; Binding; Biological Assay; Biological Process; Biomedical Research; Blood; Blood specimen; Brain; CRISPR/Cas technology; Categories; Cell model; Cells; Clinical; Clinical Research; Clustered Regularly Interspaced Short Palindromic Repeats; Cognition; Cognitive; Complex; Data; Dementia; Development; Disease; Exons; Functional disorder; Gene Expression; Genes; Genetic; Hippocampus (Brain); Homer 1; In Vitro; Individual; Induced pluripotent stem cell derived neurons; Injections; Introns; Joining Exons; Knock-out; Knockout Mice; Knowledge; Link; MAPT gene; Mediating; Messenger RNA; Molecular; Mus; Mutation; Nerve Degeneration; Neurologic; Neurons; Neurosciences; Pathogenicity; Pathology; Pathway Analysis; Pathway interactions; Penetrance; Phenotype; Protein Isoforms; Proteins; RNA Splicing; Research; Risk; Risk Factors; Role; Sampling; Senile Plaques; Time; Transcript; Transgenic Mice; Translating; Untranslated RNA; Up-Regulation; Wild Type Mouse; Work; autosomal dominant Alzheimer' s disease; base; biomarker performance; brain cell; brain tissue; circular RNA; cohort; differential expression; flexibility; frontal lobe; gene network; in vivo; knock-down; knockout animal; mRNA Precursor; molecular phenotype; mouse model; multimodality; mutation carrier; nervous system disorder; neuropathology; non-demented; novel; novel strategies; overexpression; predictive modeling; presenilin-1; presenilin-2; synaptic function; tau aggregation; tool; trafficking; trait; transcriptome; transcriptome sequencing; transcriptomics; vector

Abstract Transcriptomic studies in clinical and biomedical research have mainly focused on changes in linear transcripts to provide knowledge of the genes and co-expression networks implicated in the disease. However, very little is known about the role of circular RNAs (circRNAs) in Alzheimer's disease (AD). CircRNAs are a novel category of non-coding RNAs derived from the back-splicing and covalent joining of pre-mRNA exons and introns. We recently performed a transcriptome-wide analysis of circRNA differential expression in the brain cortex from more than 621 brain samples from two independent and large cohorts of late-onset sporadic AD cases and neuropathology-free individuals. We identified specific circRNAs, including circHOMER1, associated with AD risk and neuropathological traits. This project will use brain tissue to identify additional brain circRNAs implicated in AD from a cohort that is four-fold larger than the cohort in our previous study. We also plan to investigate differentially expressed blood circRNAs in AD cases compared with controls to determine their biomarker utility for creating new prediction models. We will establish a framework for in vitro and in vivo functional characterization of the role of circRNAs in AD. As proof of principle, we will start by defining the role of circHOMER1 in AD-related gene expression and related cellular phenotypes. We have found that circHOMER1 is highly expressed in induced pluripotent stem cell (iPSC)-derived neurons. We will use CRISPR/Cas9 to knock down circHOMER1 as well as use circHOMER1 overexpression (OE) in iPSC-derived neurons from isogenic controls and AD patient-derived neuronal cultures from pathogenic mutation carriers of APP, PSEN1, PSEN2, and MAPT genes. To determine whether circHomer1 accelerate/increase AD-related pathology in vivo, we will generate circHomer1-knockout transgenic mice and cross them with 5XFAD and MAPT-P301S mice. We will also use AAV2/9-mediated circHomer1 OE in the cortex and hippocampus of 5XFAD and MAPT-P301S mice. We recently found that seven-month-old 5XFAD mice display significant reductions of circHomer1 expression, similar to the postmortem brains of AD patients. We will restore circHomer1 levels in the hippocampus and cortex of 5XFAD and P301S mice using AAV2/9 vectors. This proposal will be the first to systematically analyze the role of brain and blood circRNAs in AD and to perform in vitro and in vivo functional studies to characterize the role of circRNAs in AD.

抽象的 临床和生物医学研究中的转录组学研究主要集中在线性转录本的变化 提供与疾病有关的基因和共表达网络的知识。然而，很少有 已知环状 RNA (circRNA) 在阿尔茨海默病 (AD) 中的作用。 CircRNA是一个新类别 源自前 mRNA 外显子和内含子的反向剪接和共价连接的非编码 RNA。我们 最近对大脑皮层中的 circRNA 差异表达进行了全转录组分析 超过 621 个大脑样本来自两个独立的大群体迟发性散发性 AD 病例 无神经病理学的个体。我们鉴定了与 AD 相关的特定 circRNA，包括 circHOMER1 风险和神经病理学特征。该项目将使用脑组织来识别其他相关的大脑 circRNA AD 患者的队列规模是我们之前研究中队列的四倍。我们也计划调查 与对照组相比，AD 病例中差异表达的血液 circRNA 以确定其生物标志物效用 用于创建新的预测模型。我们将建立一个体外和体内功能框架 circRNA 在 AD 中作用的表征。作为原则证明，我们将首先定义以下角色： circHOMER1 在 AD 相关基因表达和相关细胞表型中的作用。我们发现 circHOMER1 在诱导多能干细胞 (iPSC) 衍生的神经元中高度表达。我们将使用CRISPR/Cas9来敲 下调 circHOMER1 以及在来自同基因的 iPSC 衍生神经元中使用 circHOMER1 过表达 (OE) 对照和 AD 患者来源的神经元培养物来自 APP、PSEN1、PSEN2、 和 MAPT 基因。为了确定 circHomer1 是否会加速/增加体内 AD 相关病理，我们将 生成 circHomer1 敲除转基因小鼠，并将其与 5XFAD 和 MAPT-P301S 小鼠杂交。我们将 还在 5XFAD 和 MAPT-P301S 小鼠的皮质和海马中使用 AAV2/9 介导的 circHomer1 OE。 我们最近发现 7 个月大的 5XFAD 小鼠表现出 circHomer1 表达显着减少， 与AD患者死后的大脑相似。我们将恢复海马体和皮质中的 circHomer1 水平 使用 AAV2/9 载体的 5XFAD 和 P301S 小鼠。该提案将是第一个系统分析 脑和血液 circRNA 在 AD 中的作用，并进行体外和体内功能研究来表征 circRNA 在 AD 中的作用。