The Role of MIF in Endothelial Apoptosis

MIF 在内皮细胞凋亡中的作用

• 批准号: 6954642
• 负责人: RACHEL L DAMICO
• 金额: $ 4.57万
• 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 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6954642
• 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 是脓毒症期间循环细胞和内皮细胞中升高的一种细胞因子，在该疾病的病理生理学中发挥着关键作用。我们实验室和其他实验室的研究表明，人类 EC 对细胞凋亡刺激表现出高度抵抗力，这与抗细胞凋亡途径的上调一致。我们假设 MIF 的作用是阻止这些 EC 中的细胞凋亡。本研究的目的是通过以下目标来表征 MIF 在内皮细胞凋亡和激活中的作用：在 SA #1 中，我们将使用两种方法表征缺乏 MIF 的人肺动脉内皮细胞 (HPAEC) 的凋亡：1) 我们将使用 RNA 干扰技术下调 HPAEC 中 MIF 的产生，2) 我们将用中和方法破坏 MIF 与受体的相互作用。 抗体。然后使用缺氧、化学/再灌注、TNF 和内毒素治疗来测试表达 MIF 和 MIF 缺陷的 HPAEC 对细胞凋亡刺激的敏感性。在 SA#2 中，我们定义了 HPAEC 中对 MIF 刺激的已知和新颖的反应。最后，我们将描述 SA#3 中缺乏 MIF 时暴露的细胞凋亡途径。