Genetic models of sporadic Alzheimers Disease in the marmoset

狨猴散发性阿尔茨海默病的遗传模型

• 批准号: 10472633
• 负责人: KUO-FEN LEE
• 金额: $ 141.04万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10472633
• 关键词: Affect; Aging; Alleles; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease risk; Amino Acids; Amyloid beta-Protein; Amyloid beta-Protein Precursor; Animals; Apolipoprotein E; Apolipoproteins A; Arginine; Behavioral; Biochemical; Biological Models; Body Size; Brain; Brain Pathology; Callithrix; Clinical; Clinical Trials; Clustered Regularly Interspaced Short Palindromic Repeats; Cognitive; Complex; Cysteine; Dementia; Dementia with Lewy Bodies; Disease; Environmental Risk Factor; Exhibits; Fishes; Genes; Genetic Models; Genomics; Genotype; Goals; Guide RNA; Homeostasis; Human; Incidence; Infection; Inflammation; Inflammatory Response; Link; Lipid Binding; Lipopolysaccharides; Longevity; Magnetic Resonance Imaging; Mediating; Meta-Analysis; Metabolism; Microglia; Modeling; Molecular; Molecular Disease; Mus; Mutation; Nerve Degeneration; Neurosciences Research; Parkinson' s Dementia; Pathology; Pathway interactions; Phenotype; Point Mutation; Positioning Attribute; Positron-Emission Tomography; Predictive Value; Primates; Protein Isoforms; Proteins; Relative Risks; Reproducibility; Research; Risk; Risk Factors; Role; Signal Transduction; Structure; Synapses; Tauopathies; Testing; Threonine; Transgenic Mice; Translating; Variant; Woman; age related; apolipoprotein E-3; cerebrovascular; cognitive testing; early onset; familial Alzheimer disease; fly; gain of function; genetic linkage analysis; genetic variant; genome wide association study; glucose metabolism; immune function; lipid transport; lipidomics; mouse model; mutant; neuroimaging; nonhuman primate; null mutation; overexpression; presenilin-1; presenilin-2; protective allele; protein TDP-43; reproductive; response; risk variant; sex; social; tau Proteins; therapeutic target; tool; transgenic model of alzheimer disease; translational neuroscience

Abstract The long-term goal of this project is to develop, characterize, and validate genetically modified marmoset models of sporadic Alzheimer's Disease (AD) that will serve as tools for investigating molecular and cellular disease mechanisms, and for identifying therapeutic targets. AD is the most common cause of dementia, with the majority of cases (~95%) appearing to be sporadic, which is caused by complex interactions between multiple gene variants and environmental factors. Numerous models of AD have been developed (e.g., mice); these models are primarily focused on familial forms of AD and have enabled significant progress toward understanding AD, but they fail to recapitulate the full spectrum of molecular, cellular, behavioral, and cognitive pathologies observed in AD and provide poor predictive value when trying to translate findings to human clinical trials. Several lines of evidence suggest that marmosets may effectively bridge the gap between mice and humans for both basic and translational neuroscience research. First, marmosets and humans have very similar brain structures, cognitive/social behavioral repertoires, metabolism, and immune functions. Second, compared to other primates, marmosets have short lifespan, small body size, and high reproductive power. Finally, gene editing tools are now available to generate various types of genetically modified marmosets. Apolipoprotein E (APOE) is the strongest risk factor for late-onset, sporadic AD and also increases the age- dependent risk of monogenic familial AD and incidence of AD in women. There are three APOE alleles in humans with the APOE*ε4 allele conferring increased risk and the APOE*ε2 allele conferring decreased risk relative to the common APOE*ε3 allele. APOE isoforms differentially modulate both amyloid-β (Aβ)-dependent and Aβ-independent pathways to affect brain homeostasis, including tau-mediated neurodegeneration, microglial inflammation, lipid transport, synaptic integrity, glucose metabolism and cerebrovascular function. These three APOE alleles differ with regard to cysteine (C) and arginine (R) amino acids at positions 112 and 158 (C112/C158 in APOE2; C112/R158 in APOE3; and R112/R158 in APOE4). Several lines of evidence demonstrate that R61 is critical for APOE4-mediated AD risk. Marmoset APOE (mAPOE) is encoded by a single allele and contains the equivalent of R112 and R158, but lacks the critical R61 equivalent (it contains T instead), suggesting that it behaves like human APOE3. To test this hypothesis, mAPOE T61R mutant protein was generated to test the effect on inflammatory responses induced by lipopolysaccharides in microglial cells. The mAPOE T61R variant resulted in a more robust response compared to wild type mAPOE. Similar results were found when human APOE4 was used (APOE4>APOE3). Taken together, these preliminary results indicate that mAPOE T61R is functionally equivalent to human APOE4. Further, several pairs of CRISPR gRNAs for generating an APOE null mutation have been identified. To investigate the role of APOE in sporadic AD in the marmoset, APOE null and T61R mutant marmosets will be generated and characterized.

抽象的 该项目的长期目标是开发，表征和验证一般修改的Marmoset 散发性阿尔茨海默氏病（AD）的模型，该模型将用作研究分子和细胞的工具 疾病机制，用于识别治疗靶标。广告是痴呆症的最常见原因， 大多数病例（〜95％）似乎是零星的，这是由复杂的相互作用引起的 多个基因变体和环境因素。已经开发了许多AD模型（例如，小鼠）； 这些模型主要集中于广告的家庭形式，并能够取得重大进展 了解AD，但他们无法概括分子，细胞，行为和认知的全部光谱 在AD中观察到的病理学，并在试图将发现转化为人类时提供了差的预测价值 临床试验。有几条证据表明，摩尔果会有效地弥合小鼠之间的缝隙 和人类的基础和翻译神经科学研究。首先，摩尔果和人类非常 类似的大脑结构，认知/社会行为曲目，代谢和免疫功能。第二， 与其他素数相比，摩尔果会具有短的寿命，体积较小和繁殖能力。 最后，现在可以使用基因编辑工具来生成各种类型的一般修改的摩尔果。 载脂蛋白E（APOE）是迟发性，零星AD的强大风险因素，也会增加年龄 - 妇女中依赖性家庭广告和AD事件的依赖风险。有三个Apoe等位基因 与ApoE*ε4等位基因会议的人类增加了风险，ApoE*ε2等位基因会降低风险 相对于常见的ApoE*ε3等位基因。 APOE同工型不同地调节两个淀粉样蛋白-β（Aβ）依赖性 和非Aβ无依赖性的途径，以影响大脑稳态，包括tau介导的神经变性， 小胶质细胞注射，脂质转运，突触完整性，葡萄糖代谢和脑血管功能。 这三个APOE等位基因在半胱氨酸（C）和精氨酸（R）氨基酸方面不同，位置112和 158（APOE2中的C112/C158； APOE3中的C112/R158； APOE4中的R112/R158）。几条证据 证明R61对于APOE4介导的AD风险至关重要。 Marmoset Apoe（Mapoe）由A编码 单个等位基因，并包含R112和R158的等效物 相反），表明它的行为与人类apoe3一样。为了检验该假设，MapoE T61R突变蛋白 生成以测试小胶质细胞中脂多糖引起的炎症反应的影响。 与野生型Mapoe相比，MAPOE T61R变体产生了更强大的响应。相似的结果 在使用人apoe4时发现（apoe4> apoE3）。综上所述，这些初步结果 表明MAPOE T61R在功能上等同于人类APOE4。此外，几对CRISPR 已经确定了用于产生APOE NULL突变的GRNA。调查apoe在 将生成和表征Marmoset，Apoe Null和T61R突变型果mar虫中的零星AD。