Comprehensive Resource for the Drosophila 4th chromosome

阿尔茨海默氏症相关疾病血管性痴呆是由于年龄相关的少量血液积累所致 大脑中的血管破裂。它影响了近三分之一的70岁以上的人。任何治疗方法都不能逆转 反映血管破裂对大脑造成损害的痴呆症。更好地理解 年龄相关性脑血管虚弱的分子机制是最有可能的途径 改进诊断和痴呆症预防。本补充的目标是开发表达苍蝇的品系 遗传性血管虚弱遗传性出血与载脂蛋白E基因突变的关系 毛细血管扩张症(HHT)将被用来检验阿尔茨海默病和阿尔茨海默病的两个假说 血管性痴呆。假设是：1)产生HHT的胚系突变是随着时间的推移而概括的 通过导致年龄相关性脑血管虚弱的体细胞突变和2)HHT突变 与APOE4协同作用加速血管破裂。本副刊在母公司范围内 获奖是因为我们被资助为果蝇第四条染色体创建资源。此资源包括 4号出现每个基因功能缺失突变的新品系，表达每个基因的新品系 在第4号和新的苍蝇品系上，每4条染色体基因表达两个最接近的人类同源物。在……里面 在这份补充材料中，我们将创造出表达人类基因的新的苍蝇品系，这些品系带有APOE4和HHT突变 在一个试点项目中使用他们。亲本和补体之间的联系是5个基因中的4个 飞行员在第4条染色体上。我们已经创造了许多必要的菌株，因此预计 在一年内完成试点。首先，我们将在5个果蝇基因(Apolpp和4个)中产生功能缺失突变 与HHT有牵连的同源基因)。这些突变被设计来驱动转基因在 突变基因的天然表达模式。为了设置基线，苍蝇基因的表达对应于 用突变体来挽救突变的表型。救援实验将集中在苍蝇血液中的缺陷 模拟APOE4突变表型的脑屏障，并作为HHT的人类血管系统的类似物 突变。然后，我们通过过度表达同源人类同源基因来评估 救援程度。然后通过表达载脂蛋白E变异体和人类与HHT的同源物来检验这一假设 个体突变和组合突变。组合实验允许评估增强和 抑制互动。男性和女性的表型差异以及年龄相关性 将注意到外显率的增加。我们所产生的菌株可用于研究其他载脂蛋白E和HHT 社区对人性化苍蝇的相关疾病。我们的结果可能会刺激我们努力复制我们的 在衰老、痴呆和阿尔茨海默病的脊椎动物模型中的发现。脊椎动物模型实验可以 提供对导致血管性痴呆的分子机制的细微差别的理解 确定预测性诊断，或许还有预防性治疗。

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.