The role of CELF2 and its genetic variants in Alzheimer's disease

CELF2及其遗传变异在阿尔茨海默病中的作用

基本信息

批准号：
10334445
负责人：
Lizhen Chen
金额：
$ 45.11万
依托单位：
UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-02-01 至 2026-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10334445
关键词：
Adult Affect Age Aging Alleles Alternative Splicing Alzheimer&apos s Disease Alzheimer&apos s disease brain Alzheimer&apos s disease model Alzheimer&apos s disease pathology Alzheimer&apos s disease patient Alzheimer&apos s disease risk Animal Model Animals Autopsy Behavior Behavioral Binding Biochemical Genetics Brain Brain region Caenorhabditis elegans Clustered Regularly Interspaced Short Palindromic Repeats Cognition Data Defect Dementia Deposition Disease Elderly Environmental Risk Factor Exons Family Genes Genetic Genetic Polymorphism Genetic Screening Genomics Health High-Throughput Nucleotide Sequencing Hippocampus (Brain)Homologous Gene Human Human Genetics Immunohistochemistry Immunoprecipitation Impaired cognition Individual Induced pluripotent stem cell derived neurons Institutes Introns Knock-in Knock-out Knockout Mice Late Onset Alzheimer Disease Learning Longevity MAPT gene Mammals Measures Memory Messenger RNA Microtubules Minor Modeling Mus Mutation Nerve Degeneration Nervous system structure Neurodegenerative Disorders Neurons Pathogenesis Pathology Patients Phenotype Play Protein Isoforms Proteins Publishing RNA Splicing RNA Stability RNA-Binding Proteins Regulation Reporting Research Risk Factors Role Sampling Single Nucleotide Polymorphism Spinal Muscular Atrophy Tauopathies Techniques Testing Tissues Work brain cell calmodulin-dependent protein kinase II cell type conditional knockout crosslink crosslinking and immunoprecipitation sequencing experience genetic analysis genetic association genetic variant genome wide association study hTau Mice high risk improved induced pluripotent stem cell insight knock-down mRNA Expression member mouse model mutant neurotoxicity new therapeutic target overexpression potential biomarker presenilin-1 presenilin-2 protective effect protein expression risk variant tau Proteins tau expression therapeutic target transcriptome sequencing

项目摘要

Age is the most important risk factor for Alzheimer's disease (AD), but the occurrence of this disease is also affected by environmental factors, individual experience and genetic pre-deposition. Genetic factors are well established to play an important role in risk of AD. CELF2, an RNA binding protein that regulates alternative splicing and RNA stability, has been recently identified as a risk factor associated with AD. Polymorphisms in CELF2 are significantly associated with high-risk alleles of APOE, and the “A” allele of SNP rs2242451 is associated with reduced AD risk. CELF2 is highly expressed in the nervous system, and enhanced neuronal CELF2 expression levels have been found in various neurodegeneration models and human patients. We generated a conditional knockout mouse Celf2 allele. Our preliminary data suggest that deleting Celf2 in adult brain has beneficial effects, including improved learning and memory. We identified mRNA targets of mouse CELF2 (using CLIP-seq; cross-linking immunoprecipitation high-throughput sequencing) and found that CELF2 binds to introns around the alternatively spliced exons of a set of AD- regulated genes, including APP, MAPT (Tau), PSEN1, PSEN2, and BIN1, suggesting a key role of CELF2 in regulating alternative splicing of AD-related genes. Alternative splicing of these AD-related genes is known to regulate AD pathogenesis. For example, alternative splicing of exon 10 of the tau mRNA gives rise to protein isoforms with three (3R) or four (4R) microtubule binding repeats. Imbalances in 4R: 3R ratio alone have been reported sufficient to induce the pathogenesis of AD in a human-Tau mouse model. Taken together with the genetic association between CELF2 SNP and reduced AD risk in humans, we hypothesize that CELF2 expression is up-regulated in AD brains and loss of CELF2 in adult brains is sufficient to rescue AD-related phenotypes. In Specific Aim 1, we will test whether loss of CELF2 can suppress AD-related phenotypes in C. elegans AD models. In Specific Aim 2, we will test whether loss of CELF2 in the adult brain is protective through regulating alternative splicing using AD mouse models. In Specific Aim 3, we will test whether CELF2 expression is increased in AD brains using postmortem human samples and ask if the AD risk-reducing SNP down regulates CELF2 expression or inhibits its function using human iPSC-derived neurons. We have obtained postmortem brain samples and established a strong research team with expertise in genetics, genomics, postmortem AD brains and iPSC. Data from the proposed work will provide important mechanistic insights that go well beyond published human genetic analyses and ultimately yield new therapeutic targets for the treatment of AD.

年龄是阿尔茨海默病（AD）最重要的危险因素，但该病的发生还受环境因素、个人经历和遗传预沉积等因素的影响。 CELF2（一种调节 RNA 的结合蛋白）在 AD 风险中发挥着重要作用。选择性剪接和 RNA 稳定性最近被确定为与 AD 相关的危险因素。 CELF2 的多态性与 APOE 的高风险等位基因和 SNP 的“A”等位基因显着相关 rs2242451 与 AD 风险降低相关，CELF2 在神经系统中高表达，并且在各种神经退行性变模型中发现神经 CELF2 表达水平增强我们生成了条件敲除小鼠 Celf2 等位基因。我们发现，删除成人大脑中的 Celf2 具有有益的作用，包括改善学习和记忆力。小鼠 CELF2 的 mRNA 靶标（使用 CLIP-seq；交联免疫沉淀高通量测序），发现CELF2与一组AD-的选择性剪接外显子周围的内含子结合调节基因，包括 APP、MAPT (Tau)、PSEN1、PSEN2 和 BIN1，表明 CELF2 在已知这些AD相关基因的选择性剪接是调节AD相关基因的选择性剪接的。例如，tau mRNA 外显子 10 的选择性剪接产生蛋白质。具有三个 (3R) 或四个 (4R) 微管结合重复的亚型仅在 4R: 3R 比例中存在不平衡。据报道，足以在人 Tau 小鼠模型中诱导 AD 的发病机制。 CELF2 SNP 与人类 AD 风险降低之间的遗传关联，我们追求 CELF2 AD 大脑中的表达上调，成人大脑中 CELF2 的缺失足以挽救 AD 相关疾病在具体目标 1 中，我们将测试 CELF2 的丢失是否可以抑制 C 中 AD 相关的表型。在特定目标 2 中，我们将测试成人大脑中 CELF2 的缺失是否具有保护作用。通过使用AD小鼠模型调节选择性剪接，在Specific Aim 3中，我们将测试CELF2是否存在。使用死后人类样本，AD 大脑中的表达增加，并询问降低 AD 风险的 SNP 是否存在使用人类 iPSC 衍生的神经元下调 CELF2 表达或抑制其功能。获得死后大脑样本并建立了一支拥有遗传学专业知识的强大研究团队，基因组学、死后 AD 大脑和 iPSC 的数据将提供重要的机制。远远超出已发表的人类遗传分析的见解，并最终产生新的治疗靶点 AD的治疗。