Disease Model Development and Phenotyping Project

摘要 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，准确地模拟晚发性AD的病理 (LOAD)并为治疗开发提供预测模型。这些模型将在以下条件下生成 透明开放的知识产权条件。杰克逊实验室将进行第二现场验证 这将有助于建立负荷模型，并将确保其广泛提供并迅速传播给所有研究人员。这个 这一努力的基础是我们的创新应用程序敲入(app-ki)鼠标，它表达了人性化的Aüat 生理水平，并显示淀粉样斑块沉积(见导言)。我们将完成我们的 基础平台用于开发鼠标模型的负载，通过人性化的鼠标Tau(MAPT)轨迹 用人类Tau(MAPT)基因座的编码外显子替换小鼠Tau的编码外显子。使纯合子加倍 APP-KI和hTau(APP-KI+/+/hTau+/+)小鼠，我们将增加负荷的主要危险因素APOE4。加快步伐 分析影响负荷的其他因素，DMDPP将广泛使用CRISPR/CAS9 产生动物模型的技术，该模型表达从全基因组识别的风险因素的多态 联合研究(Gwas)，包括TREM2、PICALM、BIN1、CD2AP、ABCA7和EPHA1。的影响 当与APP-KI结合时，将确定产生负载表型的每个Gwas等位基因，并且 HTau。随后，我们将结合特定的GWAS等位基因来研究协同作用对负荷的影响 病理学。在APP-KI+/+/hTau+/+背景上整合多个GWAS多态和 将使用试管受精和胚胎移植来加速用于分析的小鼠队列的生产，而不是 标准的饲养方式。 这些负荷模型中的分子病理表型将在一个 通过一种新颖和创新的免疫学方法，使用构象，获得前所未有的详细程度 区分八种不同类型淀粉样蛋白的依赖性和聚集性特异性单抗 在人类和转基因小鼠体内沉积。还将全面确定分子表型。 通过量化神经元和小胶质细胞、突触丢失、可溶性和不溶性tau和Aü以及磷酸化标志物 陶先生。核心还将通过RNAseq和表观遗传学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。行为上的 表型的特征是高架迷宫、开阔的场地和新奇的物体识别。广博的数据 将用于比较分子病理和LOAD的分子病理，并决定将哪些线路 为进一步开发和测试生产下一代动物模型而推进。

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.