Vascular-Mediated Neuronal Cell Death Alzheimer's

描述(由申请人提供)：正在出现的数据支持在阿尔茨海默病(AD)大脑中发现具有血管生成特征的因素和过程的想法。我们已经证明，在AD中，微血管表达或释放许多炎性、促血管生成蛋白。尽管AD大脑中的促血管生成因子增加，但缺乏血管增加的证据。在我们的模型中，我们假设血管生成过程不会进展到新的血管生长，因为促血管生成因子和抗血管生成因子的失衡导致血管生成信号的中断。在这个项目中，我们验证了这样的假设，即AD微血管表达一种先天表型，这种脑内皮细胞的异常激活对AD病理的发展是重要的。目的1：验证阿尔茨海默病患者脑微血管激活但不能完成血管生成的假说，这是由于促血管生成因子和抗血管生成因子失衡导致血管生成信号中断所致。脑微血管分离自AD患者、年龄匹配的非痴呆对照组以及炎症性和非炎症性CMS疾病患者。比较分离的血管中促血管生成因子和抗血管生成因子的表达，包括凝血酶、血管内皮生长因子、内皮素-1转化生长因子-0、一氧化氮、凝血酶反应蛋白和淀粉样β蛋白。检测PI3K/Akt、p38蛋白、细胞外信号调节蛋白(ERK)、c-jun氨基末端蛋白(JNK)等信号通路的活性。脑切片免疫组织化学方法检测促血管生成蛋白和抗血管生成蛋白与A/？证词和AD病理学。目的：在阿尔茨海默病动物模型中，验证血管生成表型的获得在阿尔茨海默病病理和认知障碍发病机制中的作用。为了确定血管生成表型的获得与疾病发病之间的时间关联，在认知改变和AD病理开始之前以及疾病进展过程中的几个年龄段，测量了从AD转基因小鼠获得的分离的脑微血管中的血管生成标记物。使用抗血管生成药物评估血管生成表型和疾病进展之间的因果联系。在行为改变和AD病理发生之前给动物使用这些药物将决定抑制血管生成表型是否会影响病程。总之，来自目标1的数据显示了AD中血管生成变化的临床相关性，而目标2的结果表明了血管生成表型与疾病进展之间的因果联系，这将有力地证明AD的新治疗方法。这些结果可能非常令人兴奋，因为血管生成脑内皮细胞是一个新的、尚未探索的治疗靶点，而且几种抗血管生成药物目前正在进行第三阶段临床试验。因此，血管生成抑制剂的临床研究可以在AD患者中迅速设计和实施。

DESCRIPTION (provided by applicant): Data are emerging to support the idea that factors and processes characteristic of angiogenesis are found in the Alzheimer disease (AD) brain. We have shown that in AD microvessels express or release many inflammatory, proangiogenic proteins. Despite increases in proangiogenic factors in the AD brain, evidence for increased vascularity is lacking. In our model we hypothesize that the angiogenic process does not progress to new vessel growth because an imbalance of pro- and anti-angiogenic factors results in aborted angiogenic signaling. In this project we test the hypothesis that AD microvessels express an anqiogenic phenotype and that this abnormal activation of brain endothelial cells is important for the development of AD pathology. Aim 1: To test the hypothesis that in AD brain microvessels become activated but fail to complete angiogenesis because an imbalance of pro- and anti-angiogenic factors results in aborted angiogenic signaling. Brain microvessels are isolated from AD patients, age-matched non-demented controls, and patients with inflammatory and non-inflammatory CMS disease. Isolated vessels are compared for expression of pro- and anti-angiogenic factors including thrombin, VEGF, endothelin-1 TGF-/0, nitric oxide, thrombospondin, and amyloid beta (A¿). The activities of signaling kinases phosphatidylinositol-3 kinase (PI3K)/Akt, p38 kinase, extracellular signal-regulated kinase (ERK), and c-Jun NH2-terminal kinase (JNK) are measured. Immunohistochemistry of brain sections is used to assess the spatial correlation of pro- and anti-angiogenic proteins with A/? deposition and AD pathology. Aim 2: To test the hypothesis that acquisition of the angiogenic phenotype contributes to the pathogenesis of AD pathology and cognitive impairment in animal models of AD. To determine the temporal association between acquisition of the angiogenic phenotype and the onset of disease, markers of angiogenesis are measured in isolated brain microvessels obtained from AD transgenic mice before the onset of cognitive changes and AD pathology and at several ages during disease progression. A causal link between the angiogenic phenotype and disease progression is evaluated using antiangiogenic drugs. Administration of these drugs to animals prior to the onset of behavioral changes and AD pathology will determine whether inhibiting the angiogenic phenotype affects the course of disease. Taken together, data from Aim 1 showing the clinical relevance of angiogenic changes in AD and results from Aim 2 demonstrating a causal link between the angiogenic phenotype and disease progression would argue strongly for a new therapeutic approach in AD. These results could be very exciting because the angiogenic brain endothelial cell is a novel, unexplored therapeutic target, and several antiangiogenic drugs are currently in use in Phase III clinical trials. Thus, clinical studies with angiogenesis inhibitors could be rapidly designed and implemented in AD patients.