Centrosome Over-duplication and Blood Vessel Function

中心体过度复制与血管功能

• 批准号: 8627974
• 负责人: Erich J Kushner
• 金额: $ 5.51万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-07-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8627974
• 关键词: Abnormal Endothelial Cell; Assimilations; Behavioral; Biological Assay; Blood Vessels; Cells; Centrosome; Characteristics; DNA; Defect; Dependence; Development; Diagnostic Neoplasm Staging; Drug Targeting; Endothelial Cells; Environment; Event; Exhibits; Fibroblasts; Frequencies; Genetic; Growth; Human; Hypoxia; In Vitro; Investigation; Link; Magic; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mediating; Microtubule-Organizing Center; Modification; Mus; Neoplasms in Vascular Tissue; Nutrient; Oxygen; Pathogenesis; Phenotype; Proliferating; Recruitment Activity; Relapse; Resistance; Resistance development; Role; Signal Transduction; Solid; Solid Neoplasm; Structure; Surface; Testing; Therapeutic; Transgenic Organisms; Tumor Tissue; Tumor stage; Vascular Endothelial Growth Factors; angiogenesis; base; cell behavior; cell motility; cell type; design; insight; macrophage; monocyte; novel; novel therapeutics; prognostic; public health relevance; research study; tumor; tumor progression; tumor vascular supply

DESCRIPTION (provided by applicant): Tumor blood vessels have morphogenetic defects that impact tumor pathogenesis, primarily through the initiation of hypoxia-related signaling. Vessels found in tumors are tortuous, dilated and leaky. Limited evidence has indicated that tumor endothelial cells (ECs) have excess centrosomes, which may explain their abnormal phenotype. However, the mechanisms responsible for these cellular alterations are largely undefined. Our group recently demonstrated that excess VEGF signaling increases the frequency of centrosome over-duplication in developing blood vessels and in human ECs. Unequal centrosome numbers (>2) can have deleterious cellular consequences due to disruption of the cells microtubule-organizing center. Propagation and assimilation of "defective" ECs during angiogenic sprouting may explain, in part, the tortuous, leaky and/or chemoresistant blood vessel phenotype commonly observed in tumor tissue. This proposal aims to test the hypothesis that tumor vessel ECs are abnormal downstream of centrosome number dysregulation, and that this dysregulation contributes to the abnormal structure and function of tumor vessels. Accordingly, we will test this hypothesis in two aims. Aim #1 will determine the mechanisms and consequences of centrosome dysregulation on EC behaviors. Genetic perturbations will be introduced into human primary ECs to cause centrosome over-duplication, independent of pleiotropic proangiogenic signaling. Modified ECs will be challenged in a migratory assay and a 3D-angiogenesis assay in which they will proliferate, sprout and branch in vitro to determine the effects of centrosome dysregulation on angiogenesis. Aim #2 will determine the linkage between tumor progression and centrosome duplication in ECs of tumor vessels. Using a novel transgenic approach to mark EC DNA, I will analyze the endothelial compartment in mouse mammary tumors for centrosome abnormalities to be correlated with tumor stage and invasiveness to determine the effects of dysregulation of centrosome duplication on tumor progression. Additionally, tumor and normal ECs will be isolated and cultured ex vivo and challenged to the same functional assays in aim #1. Information from this investigation will illuminate key early events (dysregulation of centrosome duplication) in tumor blood vessel angiogenesis, and show how these events link to abnormal vessel development and tumor progression.

描述（由申请人提供）：肿瘤血管具有影响肿瘤发病机理的形态发生缺陷，主要是通过缺氧相关信号传导的开始。在肿瘤中发现的血管曲折，扩张和漏水。有限的证据表明，肿瘤内皮细胞（EC）具有过量的中心体，这可以解释其异常表型。但是，负责这些细胞改变的机制在很大程度上是不确定的。我们的小组最近证明，过量的VEGF信号传导增加了发育血管和人类EC中的中心体过度杀取率的频率。由于细胞微管 - 组织中心的破坏，不等的中心体数（> 2）可能会产生有害的细胞后果。血管生成期间“有缺陷” EC的传播和同化可能部分解释了在肿瘤组织中通常观察到的曲折，漏水和/或化学耐药性血管表型。该建议旨在检验以下假设：肿瘤血管EC是中心体数量失调的下游异常，并且这种失调有助于肿瘤血管的异常结构和功能。因此，我们将以两个目标来检验这一假设。目标＃1将确定中心体失调对EC行为的机制和后果。遗传扰动将被引入人类原发性ECS，以引起中心体过度检验，而与多效性促肌动型信号无关。修改的EC将在迁移分析和3D血管生成测定中受到挑战，其中它们将在体外增殖，发芽和分支，以确定中心体失调对血管生成的影响。 AIM＃2将确定肿瘤血管EC中肿瘤进展与中心体重复之间的联系。使用一种新型的转基因方法来标记EC DNA，我将分析小鼠乳腺肿瘤中的内皮区室的中心体异常，以与肿瘤阶段和侵入性相关，以确定中心体重复对肿瘤进展的失调作用。此外，肿瘤和正常EC将被分离和培养的离体，并在AIM＃1中对相同功能测定法进行挑战。该研究的信息将阐明肿瘤血管血管生成中的关键早期事件（中心体重复的失调），并显示这些事件与异常血管发育和肿瘤进展如何联系起来。