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.

摘要 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 以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是一个新的类别 来自前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 最近进行了一项全转录组的分析，从更多的大脑皮层中的circRNA差异表达， 来自两个独立的大型迟发性散发性AD病例队列的621份脑样本， 没有神经病理学的人我们鉴定了与AD相关的特定circRNA，包括circHOMER 1， 风险和神经病理学特征。该项目将使用脑组织来识别其他涉及的脑circRNA 在AD中，来自比我们先前研究中的队列大四倍的队列。我们还计划调查 与对照相比，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。我们将建立一个框架，在体外和体内功能 CircRNA在AD中的作用的表征。作为原则的证明，我们将首先定义 circHOMER 1在AD相关基因表达和相关细胞表型中的作用。我们发现circHOMER 1 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circHOMER 1以及在来自同基因的iPSC衍生的神经元中使用circHOMER 1过表达（OE）。 对照和来自APP，PSEN 1，PSEN 2， MAPT基因。为了确定circHomer 1是否在体内加速/增加AD相关病理，我们将 产生circHomer 1敲除转基因小鼠，并将它们与5XFAD和MAPT-P301 S小鼠杂交。我们将 还在5XFAD和MAPT-P301 S小鼠的皮质和海马中使用AAV 2/9介导的circHomer 1 OE。 我们最近发现，7个月大的5XFAD小鼠显示circHomer 1表达显著减少， 类似于AD患者死后的大脑。我们将恢复海马体和皮层中的circHomer 1水平 使用AAV 2/9载体的5XFAD和P301 S小鼠的免疫组化。该提案将首次系统分析 脑和血液circRNA在AD中的作用，并进行体外和体内功能研究，以表征 CircRNA在AD中的作用