Comprehensive Resource for the Drosophila 4th chromosome

果蝇第四染色体综合资源

基本信息

批准号：
10491507
负责人：
STUART J NEWFELD
金额：
$ 31.17万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10491507
关键词：
ANK2 gene Administrative Supplement Advisory Committees Affect Age Age related macular degeneration Aging Alleles Alzheimer&apos s Disease Alzheimer&apos s disease risk Ankyrins Apolipoprotein E Arizona Award Basic Science Biological Biological Assay Biological Models Biology Blood - brain barrier anatomy Blood Vessels Brain Brain hemorrhage Cells Cerebrovascular system Chromosomes Clone Cells Clustered Regularly Interspaced Short Palindromic Repeats Code Collection Communities Complementary DNA Defect Dementia Deposition Development Diagnostic Disease Drosophila genus Drosophila melanogaster Endothelial Cells Endothelium Engineering Extravasation Family Female Funding Gene Expression Regulation Gene Proteins Genes Genetic Recombination Germ Lines Germ-Line Mutation Goals Grant Health Hereditary hemorrhagic telangiectasia Homologous Gene Human Impairment Individual Inherited Ischemic Stroke Lead Light Lipids Long QT Syndrome Modeling Molecular Molecular Analysis Molecular Genetics Mutation Neurons Parents Pattern Penetrance Persons Phenotype Pilot Projects Prevention Preventive Proteins RNA Reporter Genes Research Personnel Resources Rupture Scientist Somatic Mutation Stroke Testing Therapeutic Time Tissues United States National Institutes of Health Untranslated RNA Variant Vascular Cognitive Impairment Vascular Dementia Vascular Diseases age related analog cerebrovascular combinatorial experimental study fly frontier gain of function genetic analysis improved innovation loss of function loss of function mutation male mutant overexpression parent grant transgene expression translational applications

项目摘要

The Alzheimer's related disease vascular dementia is due to an age-dependent accumulation of small blood vessel ruptures in the brain. It affects almost a third of people over age 70. There is no treatment that reverses the dementia that reflects the damage to the brain caused by broken vessels. A better understanding of the molecular mechanisms underlying age associated cerebral blood vessel weakness is the most likely path to improved diagnostics and dementia prevention. The goal of this supplement is develop fly strains expressing variants of ApoE and mutations involved in the inherited blood vessel weakness Hereditary Hemorrhagic Telangiectasia (HHT) that will be employed to test two hypotheses for the causes of Alzheimer's disease and vascular dementia. The hypotheses are: 1) that germline mutations generating HHT are recapitulated over time by somatic mutations leading to age associated brain blood vessel weakness and 2) that HHT mutations synergyze with APOE4 to accelerate blood vessel ruptures. This supplement is within the scope of the parent award since we are funded to create a resource for the Drosophila 4th chromosome. This resource includes new fly strains with loss of function mutations for each gene on the 4th, new fly strains expressing each gene on the 4th and new fly strains expressing the two closest human homologs for each 4th chromosome gene. In this supplement we will create new fly strains expressing human genes with APOE4 and HHT mutations and employ them in a pilot project. The connection between the parent and supplement is that 4 of the 5 genes in the pilot are on the 4th chromosome. We have already created many of the necessary strains and thus expect to complete the pilot in one year. First we will create loss of function mutations in 5 fly genes (Apolpp and four genes with homologs implicated in HHT). These mutations are engineered to drive transgene expression in the mutant gene's native expression pattern. To set a baseline, expression of the fly gene corresponding to the mutant is employed to rescue the mutant phenotype. Rescue experiments will focus on defects in the fly blood brain barrier to mimic an APOE4 mutant phenotype and as an analog to the human vasculature for HHT mutations. We then humanize the mutant flies by overexpressing the cognate human homolog to evaluate the degree of rescue. The hypothesis is then tested by expressing ApoE variants and human homologs with HHT mutations individually and in combinations. Combinatorial experiments allow assessment of enhancing and suppressing interactions. Phenotypic differences between males and females as well as age-dependent increases in penetrance will be noted. The strains we generate can be employed to study other ApoE and HHT related diseases in humanized flies by the community. Our results will likely stimulate efforts to replicate our findings in vertebrate models of aging, dementia and Alzheimer's disease. Vertebrate model experiments could provide the nuanced understanding of the molecular mechanisms underlying vascular dementia necessary for identifying predictive diagnostics and perhaps preventive therapeutics.

阿尔茨海默氏症相关疾病血管性痴呆是由于年龄依赖性的小血液的积累血管在大脑中破裂。它影响了70岁以上的几乎三分之一的人。没有治疗可以逆转反映骨折造成的大脑损害的痴呆症。更好地理解与年龄相关的分子机制相关的脑血管弱点是最可能的途径改善诊断和预防痴呆症。该补充的目的是发展蝇菌表达 APOE的变体和遗传血管无力遗传性出血的突变 telangictasia（HHT）将用于检验两个假设，以实现阿尔茨海默氏病和血管痴呆。假设是：1）随着时间的推移，将产生HHT的种系突变概括通过导致年龄相关的脑血管弱点的体细胞突变和2）HHT突变与APOE4的Synergyze加速血管破裂。这种补充在父母的范围内由于我们被授予为果蝇第四染色体创建资源的奖项。此资源包括新的蝇菌株在第四个基因的功能突变丧失，表达每个基因的新蝇菌株在第四和新的蝇菌株中，每个第四染色体基因表达了两个最接近的人类同源物。在这种补充剂我们将创建新的苍蝇菌株，以apoe4和hht突变表达人类基因，以及将它们雇用在试点项目中。父母和补充之间的联系是5个基因中的4个飞行员在第四个染色体上。我们已经创造了许多必要的菌株，因此期望在一年内完成飞行员。首先，我们将在5个苍蝇基因（ApoLpp和4个）中创建功能突变的丧失与HHT有关的同源物基因）。这些突变经过设计以驱动转基因表达突变基因的天然表达模式。为了设定基线，蝇基因的表达与突变体用于拯救突变表型。救援实验将集中于蝇血中的缺陷模仿APOE4突变体表型的脑屏障，并作为HHT人类脉管系统的类似物突变。然后，我们通过过表达同志人同源物来评估突变体蝇来使突变体苍蝇人性化救援程度。然后，通过用HHT表达APOE变体和人类同源物来检验该假设单独和组合突变。组合实验允许评估增强和抑制互动。男性和女性之间的表型差异以及年龄依赖性将注意到外观的增加。我们产生的菌株可用于研究其他APOE和HHT 社区中人性化果蝇中的相关疾病。我们的结果可能会激发复制我们的努力衰老，痴呆和阿尔茨海默氏病的脊椎动物模型中的发现。脊椎动物模型实验可以提供对需要血管痴呆所必需的分子机制的细微理解识别预测性诊断，也许可以预防治疗。