Molecular endophenotypes of H1 and H2 MAPT haplotypes

H1 和 H2 MAPT 单倍型的分子内表型

• 批准号: 9762819
• 负责人: Karen H Ashe
• 金额: $ 19.25万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-15 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762819
• 关键词: Affect; Age; Aging; American; Animal Model; Biological; Biological Assay; Chronic; Development; Disease; Etiology; Gene Cluster; Genes; Genetic Polymorphism; Genome; Goals; Hand; Haplotypes; Human; Knowledge; Late Onset Alzheimer Disease; Location; MAPT gene; Measures; Medical; Messenger RNA; Methodology; Modeling; Molecular; Molecular Target; Mus; Odds Ratio; Pathologic; Pathway interactions; Pattern; Progressive Supranuclear Palsy; Proteins; Reporting; Risk; Risk Factors; Structure; Tauopathies; Untranslated RNA; Variant; age related; disorder risk; endophenotype; gene replacement; genetic risk factor; genetic variant; genome wide association study; high risk; in vivo Bioassay; insight; mRNA Expression; mouse genome; mouse model; preservation; protective effect; protein expression; tau Proteins; tau dysfunction; tau phosphorylation; therapy development

Project Summary Our long-term goal is to understand the biological mechanisms underlying GWAS hits in LOAD and other tauopathies. The objective of our current proposal is to develop mouse models in which the entire 190 Kb human MAPT gene precisely replaces the complete 157 Kb mouse Mapt gene, and to use these models to compare the molecular endophenotypes of tau in young and old mice expressing H2 or H1 MAPT haplotypes. Our rationale is that the non-coding variants that define the H1 and H2 haplotypes are present in MAPT but not Mapt, necessitating the incorporation of the full MAPT gene. In addition, because we have found that the location of MAPT in the mouse genome affects its expression, we will preserve the basic structural configuration of Chr17q21.31 in which MAPT resides, and maintain its relationship relative to other genes in the gene cluster that mice and humans share. These studies will be significant because they will constitute the first in vivo bioassays of non-coding polymorphisms, paving the way for future research on GWAS hits in not only LOAD, but also other chronic medical disorders. Our overarching hypothesis is that differences in the progression and pattern of tau mRNA and protein expression underlie the variations in risk associated with H2 and H1 MAPT haplotypes. We have in-hand the first MAPT Gene-Replacement (GR) mouse line, in which the 190kb human H2 MAPT precisely replaces the 157 Kb Mapt locus, and will use the same methodology to create a precisely matched line in which the H1 MAPT replaces Mapt. We will measure and compare mRNA, protein and post-translational modifications of tau in young and old GR1 and GR2 mice. Upon completion of these studies, we will have created and characterized two lines of mice in which Mapt is precisely replaced by H1 or H2 variants of MAPT. We predict that the distinct polymorphisms in H1 and H2 MAPT will suffice to alter the molecular endophenotype of tau in GR1 relative to GR2 mice, and that these differences will increase with age.

项目摘要 我们的长期目标是了解LOAD中GWAS命中的生物学机制， 其他Tau病我们目前建议的目标是开发小鼠模型， 完整的190 Kb人MAPT基因精确地替换完整的157 Kb小鼠Mapt基因， 使用这些模型来比较表达tau蛋白的年轻小鼠和老年小鼠中tau蛋白的分子内表型， H2或H1 MAPT单倍型。我们的理由是，定义H1和H2的非编码变体 单倍型存在于MAPT中，但不存在于Mapt中，这就需要整合完整的MAPT基因。在 此外，因为我们已经发现MAPT在小鼠基因组中的位置影响其在小鼠中的表达。 表达，我们将保留MAPT所在的Chr17q21.31的基本结构构型， 并保持其相对于小鼠和人类共有的基因簇中其他基因的关系。 这些研究将是重要的，因为它们将构成第一个非编码的体内生物测定。 多态性，铺平了道路，为未来的研究GWAS命中不仅在负荷，而且在其他 慢性疾病我们的总体假设是，在进展和 Tau mRNA和蛋白表达模式是H2和H1相关风险变化的基础 MAPT单倍型。 我们已经拥有了第一个MAPT基因置换（GR）小鼠系，其中190kb的人H2 MAPT精确地取代了157 Kb的Mapt轨迹，并将使用相同的方法来创建一个 精确匹配的行，其中H1 MAPT取代了Mapt。我们将测量和比较mRNA， 蛋白质和翻译后修饰的tau蛋白在年轻和老年GR1和GR2小鼠。 在完成这些研究后，我们将创建并表征两个品系的小鼠，其中 Mapt被MAPT的H1或H2变体精确取代。我们预测，不同的多态性， H1和H2 MAPT将足以改变GR 1相对于GR 2小鼠中tau的分子内表型， 这些差异会随着年龄的增长而增加。