The Role of MIF in Endothelial Apoptosis

MIF 在内皮细胞凋亡中的作用

• 批准号: 6835813
• 负责人: RACHEL L DAMICO
• 金额: $ 5.05万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6835813
• 关键词: RNA interference; apoptosis; biological signal transduction; enzyme linked immunosorbent assay; flow cytometry; fluorescence microscopy; inflammation; macrophage; migration inhibition factor; mitogen activated protein kinase; neutralizing antibody; pathologic process; postdoctoral investigator; protein protein interaction; pulmonary artery; receptor; septicemia; vascular endothelium

DESCRIPTION (provided by applicant): Endothelial cell (EC) injury is a cardinal feature of sepsis. MIF, a cytokine elevated both in the circulation and endothelial cells during sepsis, plays a critical role in the pathophysiology of this disease. Studies in our lab and others demonstrate that human EC display a high resistance to apoptotic stimuli consistent with the upregulation of anti-apoptotic pathways. We hypothesize that MIF functions to block apoptosis in these EC. The goal of this study is to characterize the role of MIF in endothelial cell apoptosis and activation via the following aims: In SA #1 we will characterize apoptosis in human pulmonary artery endothelial cells (HPAEC) deficient in MIF using two approaches 1) we will downregulate MIF production in HPAEC using RNA interfering technology, and 2) we will disrupt MIF-receptor interactions with neutralize antibodies. The susceptibility of MIF-expressing and MIF-deficient HPAEC to apoptotic stimuli will then be tested using hypoxia, chemic/reperfusion, TNF, and endotoxin treatment. In SA#2 we define known and novel responses to MIF stimulation in HPAEC. Finally, we will characterize the apoptotic pathways exposed in the absence of MIF in SA#3.

描述（由申请人提供）：内皮细胞（EC）损伤是脓毒症的主要特征。MIF是一种在脓毒症期间在循环和内皮细胞中均升高的细胞因子，在这种疾病的病理生理学中起关键作用。我们实验室和其他实验室的研究表明，人EC对凋亡刺激具有高抗性，这与抗凋亡途径的上调一致。我们推测，MIF的功能，以阻止这些EC的凋亡。本研究的目的是通过以下目的来表征MIF在内皮细胞凋亡和活化中的作用：在SA #1中，我们将使用两种方法来表征缺乏MIF的人肺动脉内皮细胞（HPAEC）中的凋亡：1）我们将使用RNA干扰技术下调HPAEC中的MIF产生，以及2）我们将用中和抗体破坏MIF-受体相互作用。然后使用缺氧、化学/再灌注、TNF和内毒素处理来测试MIF表达和MIF缺陷的HPAEC对凋亡刺激的敏感性。在SA#2中，我们定义了HPAEC中对MIF刺激的已知和新反应。最后，我们将表征在SA#3中不存在MIF的情况下暴露的凋亡途径。