Disease Model Development and Phenotyping Project

疾病模型开发和表型分析项目

• 批准号: 10250343
• 负责人: GRANT R MACGREGOR
• 金额: $ 225.6万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10250343
• 关键词: Age; Alleles; Alzheimer' s disease model; Alzheimer' s disease pathology; Amyloid; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Animal Model; Animals; Binding; Biochemistry; Bioinformatics; Biological Markers; Breeding; CRISPR/Cas technology; Clinical Trials; Code; Cognitive; Communities; Complex; Control Animal; Data; Deposition; Development; Diffusion Magnetic Resonance Imaging; Disease; Disease model; EPHA1 gene; Electrophysiology (science); Embryo Transfer; Ensure; Epigenetic Process; Etiology; Excision; Exhibits; Exons; Fertilization in Vitro; Foundations; Gene Expression Profile; Gene Expression Profiling; Gene Transfer Techniques; Genes; Genetic Polymorphism; Goals; Histologic; Human; Immunologics; Impaired cognition; Intellectual Property; Knock-in; Knock-in Mouse; Late Onset Alzheimer Disease; Mediating; Methodology; Microglia; Modeling; Molecular; Molecular Conformation; Monoclonal Antibodies; Mouse Strains; Mus; Mutation; Neurodegenerative Disorders; Neurofibrillary Tangles; Neurons; Pathogenesis; Pathogenicity; Pathologic; Pathology; Pharmaceutical Preparations; Phenotype; Physiological; Preclinical Testing; Presenile Alzheimer Dementia; Primary Cell Cultures; Process; Production; Property; Reporting; Research; Research Personnel; Resources; Risk Factors; Senile Plaques; Site; Structure; Synapses; TREM2 gene; Testing; The Jackson Laboratory; Transgenic Mice; Validation; abeta accumulation; base; behavioral phenotyping; cognitive ability; cognitive benefits; cohort; data management; epigenetic marker; gamma secretase; gene product; genetic risk factor; genome wide association study; hTau Mice; human disease; innovation; lipidomics; metabolomics; model development; molecular pathology; molecular phenotype; mouse development; mouse genome; mouse model; neuroimaging; next generation; novel; object recognition; overexpression; predictive modeling; presenilin; risk variant; sperm cryopreservation; tau Proteins; tau aggregation; tau mutation; tau-1; therapeutic development; therapeutic effectiveness; therapeutic evaluation; tool; transcriptome sequencing

ABSTRACT The goal of the Disease Model Development and Phenotyping Project (DMDPP) is to produce and characterize the next generation of animal models for AD that accurately model the pathology of late onset AD (LOAD) and to provide predictive models for therapeutic development. These models will be generated under transparent and open intellectual property conditions. The Jackson Laboratory will conduct 2nd-site validation of LOAD models and will ensure their broad availability and rapid dissemination to all researchers. The foundation of this endeavor is our innovative APP knock-in (APP-KI) mouse that expresses humanized Aß at physiological levels, and which exhibits amyloid plaque deposition (see introduction). We will complete our base platform for development of mouse models of LOAD, by humanizing the mouse Tau (Mapt) locus by replacing coding exons of mouse Tau with those from the human Tau (MAPT) locus. To double homozygous APP-KI and hTau (APP-KI+/+/hTau+/+) mice, we will add the major risk factor for LOAD, APOE4. To expedite analysis of additional factors influencing LOAD, the DMDPP will make extensive use of CRISPR/Cas9 technology to generate animal models that express polymorphisms in risk factors identified from genome wide association studies (GWAS), including TREM2, PICALM, BIN1, CD2AP, ABCA7 and EPHA1. The effect of each GWAS allele in generating a LOAD phenotype will be determined when combined with APP-KI, and hTau. Subsequently we will combine specific GWAS alleles to investigate for synergistic effects on LOAD pathology. The incorporation of multiple GWAS polymorphisms on the APP-KI+/+/hTau+/+ background and production of cohorts of mice for analysis will be accelerated using IVF and embryo transfer instead of standard breeding. The molecular pathological phenotypes in these LOAD models will be characterized at an unprecedented level of detail through a novel and innovative immunological approach, using conformation dependent and aggregation specific monoclonal antibodies that distinguish eight different types of amyloid deposits in humans and transgenic mice. The molecular phenotype will also be comprehensively determined by quantifying neurons and microglia, synaptic loss, soluble and insoluble tau and Aß and markers of phospho tau. The core will also characterize the gene expression profile of the models by RNAseq and epigenetic markers, as well as structural, functional, and diffusion magnetic resonance imaging. The behavioral phenotype will be characterized by elevated maze, open field, and novel object recognition. The extensive data generated will be used to compare the molecular pathology to that of LOAD and to decide which lines to advance for further development and testing to produce the next generation of animal models.

抽象的 疾病模型开发和表型项目 (DMDPP) 的目标是生产和 描述下一代 AD 动物模型的特征，准确模拟迟发性 AD 的病理学 （负载）并为治疗开发提供预测模型。这些模型将在下生成 透明、开放的知识产权条件。杰克逊实验室将进行第二次现场验证 LOAD 模型，并将确保其广泛可用性并快速传播给所有研究人员。这 这项努力的基础是我们创新的 APP 敲入 (APP-KI) 小鼠，它在 生理水平，并表现出淀粉样斑块沉积（见简介）。我们将完成我们的 通过将小鼠 Tau (Mapt) 基因座人性化，用于开发 LOAD 小鼠模型的基础平台 将小鼠 Tau 的编码外显子替换为来自人类 Tau (MAPT) 位点的编码外显子。为双纯合 APP-KI 和 hTau (APP-KI+/+/hTau+/+) 小鼠，我们将添加 LOAD 的主要危险因素 APOE4。加快 分析影响 LOAD 的其他因素，DMDPP 将广泛使用 CRISPR/Cas9 生成动物模型的技术，该模型表达从全基因组中识别出的危险因素的多态性 关联研究 (GWAS)，包括 TREM2、PICALM、BIN1、CD2AP、ABCA7 和 EPHA1。的效果 与 APP-KI 结合时，将确定生成 LOAD 表型的每个 GWAS 等位基因，并且 hTau。随后我们将结合特定的 GWAS 等位基因来研究对 LOAD 的协同效应 病理。 APP-KI+/+/hTau+/+ 背景上多个 GWAS 多态性的整合 使用体外受精（IVF）和胚胎移植（而不是胚胎移植）将加速用于分析的小鼠群体的生产 标准育种。 这些 LOAD 模型中的分子病理表型将在 通过新颖和创新的免疫学方法，使用构象，获得前所未有的细节水平 依赖性和聚集特异性单克隆抗体，可区分八种不同类型的淀粉样蛋白 人类和转基因小鼠中的沉积物。分子表型也将被综合确定 通过量化神经元和小胶质细胞、突触损失、可溶性和不溶性 tau 和 Aß 以及磷酸标记物 头。该核心还将通过 RNAseq 和表观遗传来表征模型的基​​因表达谱 标记物，以及结构、功能和扩散磁共振成像。行为方面 表型的特点是高架迷宫、开放场地和新物体识别。广泛的数据 生成的结果将用于将分子病理学与 LOAD 的分子病理学进行比较，并决定要选择哪些细胞系 进一步开发和测试以生产下一代动物模型。