Interferon Alpha: The Link Between Lupus and Endothelial Dysfunction

干扰素α：狼疮与内皮功能障碍之间的联系

• 批准号: 8704712
• 负责人: Joy N Jones Buie
• 金额: $ 3.52万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 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 2015-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8704712
• 关键词: Address; African American; American; Apoptosis; Atherosclerosis; Attenuated; Autoimmune Diseases; Binding; Biological Assay; Biological Availability; Biological Markers; Biology; Biometry; Blood Circulation; Blood Vessels; Cardiovascular Diseases; Cardiovascular system; Cell Culture Techniques; Cell Proliferation; Cell physiology; Cells; Chemiluminescence assay; Chronic; Complement; Cyclic GMP; Data; Development; Disease; Endothelial Cells; Endothelium; Enzymes; Ethics; Event; Foundations; Free Radicals; Functional disorder; Genetic Transcription; Goals; Grant; Human; Immune response; Immunoblotting; Immunology; In Vitro; Incidence; Infection; Inflammation; Inflammatory; Interferon Type I; Interferon-alpha; Interferons; Journals; Laboratories; Lead; Learning; Link; Luciferases; Lupus; Measures; Mediating; Medicine; Microscopy; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Nitrates; Nitric Oxide; Nitrites; Output; Pathway interactions; Patients; Peer Review; Pharmacology; Phenotype; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Phosphotransferases; Plasmids; Population; Production; Promoter Regions; Protein phosphatase; Proto-Oncogene Proteins c-akt; Publications; Reporting; Research Personnel; Rheumatology; Role; Scientist; Series; Serine; Signal Pathway; Signal Transduction; Site; Societies; Stem cells; Students; Systemic Lupus Erythematosus; Techniques; Testing; Therapeutic; Time; Training; Transcript; Umbilical vein; Vascular Diseases; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; Work; Writing; base; cell injury; college; cytokine; endothelial dysfunction; human NOS3 protein; improved; inhibitor/antagonist; mRNA Stability; meetings; mortality; novel; patient population; prevent; promoter; protein phosphatase inhibitor-1; public health relevance; repaired; success; symposium; transcription factor; vascular endothelial dysfunction

DESCRIPTION (provided by applicant): Accelerated atherosclerosis is initiated by severe vascular endothelial dysfunction (VED), a phenotype common amongst systemic lupus erythematosus (SLE) patients. Although causes of VED are multifactorial, all pathways converge on the diminished activity of endothelial nitric oxide synthase (eNOS) expression and loss of NO bioavailability important for blood vessel protection. Recent studies indicate that interferon (IFN) -alpha contributes to the development of VED in SLE subjects by impairing endothelial repair mechanisms however, studies are missing that indicate a role for IFN-alpha in vascular damage. We recently showed that treatment of human aortic and umbilical vein endothelial cells (HAECs/HUVECs) with IFN-alpha leads to decreased eNOS expression and phosphorylation. Therefore, the purpose of the proposed study is to tease out mechanisms responsible for type I interferon-mediated inhibition of eNOS expression and activation in an in vitro HUVEC/HAEC cell culture model. We intend to test our hypothesis that IFN-alpha causes diminished eNOS expression/activity by 1) decreasing transcription factor interaction with the eNOS promoter region and by 2) causing increased protein phosphatase 2A activation and/or decreased Akt (protein kinase B) expression. To test this hypothesis we will address the following specific aims: first analyze changes in eNOS promoter region activity, mRNA stability, and transcription factor binding. Secondly, we will quantify changes in different phosphatases and kinases that may attribute to changes in eNOS activation. Finally, we will measure NO and cyclic guanosine monophosphate (cGMP) levels following IFN-alpha treatment to ascertain its impact on functional eNOS outputs. This series of studies are significant because they will provide ample evidence for the role of type I IFNs in compromising endothelial function. The proposed studies accompanied by robust coursework, scientific meeting and seminar presentations, and interactions with other scientist will provide a training plan that will lead tothe applicant's success as an academician. Coursework will address advanced topics in immunology and vascular disease, complementing previous courses including biostatistics, grant writing, scientific ethics, and pharmacology. The student will also work with NO expert, Dr. Philip Shaul to learn techniques currently not available in the laboratory and present work at conferences including the American College of Rheumatology Conference, Society for Free Radical Biology and Medicine Conference, and one of the many Gordon Conferences. Finally, the student will submit work for publication in peer-reviewed journals.

描述（由申请人提供）：严重的血管内皮功能障碍（VED）引发加速动脉粥样硬化，这是系统性红斑狼疮（SLE）患者中常见的一种表型。尽管 VED 的原因是多因素的，但所有途径均集中于内皮型一氧化氮合酶 (eNOS) 表达活性的降低以及对血管保护至关重要的 NO 生物利用度的丧失。最近的研究表明，干扰素 (IFN)-α 通过损害内皮修复机制，促进 SLE 受试者 VED 的发展，然而，缺乏表明 IFN-α 在血管损伤中发挥作用的研究。我们最近发现，用 IFN-α 治疗人主动脉和脐静脉内皮细胞 (HAEC/HUVEC) 会导致 eNOS 表达和磷酸化降低。因此，本研究的目的是在体外 HUVEC/HAEC 细胞培养模型中梳理出 I 型干扰素介导的 eNOS 表达和激活抑制的机制。我们打算检验我们的假设，即 IFN-α 通过 1) 减少转录因子与 eNOS 启动子区域的相互作用，以及 2) 导致蛋白磷酸酶 2A 激活增加和/或 Akt（蛋白激酶 B）表达减少，从而导致 eNOS 表达/活性减少。为了检验这一假设，我们将实现以下具体目标：首先分析 eNOS 启动子区域活性、mRNA 稳定性和转录因子结合的变化。其次，我们将量化不同磷酸酶和激酶的变化，这些变化可能归因于 eNOS 激活的变化。最后，我们将测量 IFN-α 治疗后的 NO 和环磷酸鸟苷 (cGMP) 水平，以确定其对功能性 eNOS 输出的影响。这一系列研究意义重大，因为它们将为 I 型干扰素在损害内皮功能中的作用提供充分的证据。拟议的研究伴随着强大的课程作业、科学会议和研讨会演示以及与其他科学家的互动，将提供一个培训计划，使申请人成为一名成功的院士。课程将讨论免疫学和血管疾病的高级主题，补充之前的课程，包括生物统计学、资助写作、科学伦理和药理学。学生还将与 NO 专家 Philip Shaul 博士合作，学习实验室目前无法使用的技术，并在美国风湿病学会会议、自由基生物学和医学学会会议以及众多戈登会议之一等会议上展示工作成果。最后，学生将提交作品以在同行评审期刊上发表。