Centrosome Mis-Regulation and Blood Vessel Function

描述（由申请人提供）：中心体使细胞中的微管成核，并在间期通过微管组织中心（MTOC）和有丝分裂期间通过纺锤体组织微管。中心体复制通常被严格调节，每个细胞周期发生一次且仅发生一次，并且这种调节的丧失导致肿瘤细胞中过量的中心体、遗传不稳定性和非整倍性。我们最近首次发现，暴露于VEGF-A信号升高的血管中的内皮细胞（EC）具有过量的中心体，通过Akt，MEK/ERK和Cdk 2/细胞周期蛋白E介导。中心体过量被预测会扰乱细胞骨架并导致细胞行为的改变，尽管这种联系尚未在细胞或组织中建立;它还被预测会产生染色体不稳定性和非整倍性，导致染色体畸变。 从常规到EC的深刻变化。这项工作将测试的假设，EC中的过量中心体扰动血管功能，通过有丝分裂和非有丝分裂的扰动，并检查机制，导致中心体失调后血管生成因子的刺激。这一高度创新的假说将被测试与三个目标，解决的问题：1）是什么输入和机制，导致EC中心体失调？2)过量的中心体对EC行为和萌发有什么影响？（3）在体血管EC中过量中心体的结果是什么？我们将利用基于细胞的血管形成模型和体内小鼠发育和疾病模型来解决这些问题。这项工作的意义将是揭示发育中血管对中心体介导的EC功能障碍和遗传不稳定性的易感性，预计这将以新的方式促进血管功能障碍，并导致新的和独特的治疗靶点。

DESCRIPTION (provided by applicant): Centrosomes nucleate microtubules in cells, and organize them via a microtubule organizing center (MTOC) during interphase and via spindles during mitosis. Centrosome duplication is normally tightly regulated to occur once and only once per cell cycle, and loss of this regulation leads to excess centrosomes, genetic instability, and aneuploidy in tumor cells. We recently showed, for the first time, that endothelial cells (EC) in blood vessels exposed to elevated VEGF-A signaling have excess centrosomes, mediated through Akt, MEK/ERK, and Cdk2/cyclin E. Centrosome excess is predicted to perturb the cytoskeleton and result in alterations in cellular behaviors, although this link has not been made in cells or tissues; it also is predicted to produce chromosome instability and aneuploidy, leading to EC with profound changes from the norm. This work will test the hypothesis that excess centrosomes in EC perturb vessel function via both mitotic and non-mitotic perturbations, and examine the mechanisms that lead to centrosome mis-regulation upon angiogenic factor stimulation. This highly innovative hypothesis will be tested with three aims that address the questions: 1) What are the inputs and mechanisms that lead to EC centrosome mis- regulation? 2) What are the consequences of excess centrosomes to EC behaviors and sprouting? and 3) what are the outcomes of excess centrosomes in EC of vessels in vivo? We will utilize cell- based models of blood vessel formation and in vivo mouse developmental and disease models to address the questions. The significance of this work will be to reveal susceptibility in developing vessels for centrosome-mediated EC dysfunction and genetic instability, which is predicted to contribute to blood vessel dysfunction in novel ways, and lead to new and distinct therapeutic targets.