Role of a novel IL18 binding protein in atherosclerosis

新型 IL18 结合蛋白在动脉粥样硬化中的作用

基本信息

批准号：
8882740
负责人：
GUO-PING SHI
金额：
$ 42.04万
依托单位：
BRIGHAM AND WOMEN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-08-21 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8882740
关键词：
Adhesions Affect Affinity Age Aldosterone Angiotensin II Aorta Apolipoprotein E Apoptosis Area Arterial Fatty Streak Atherosclerosis Binding Biological Models Blood Vessels Bone Marrow Cells CCL2 gene COS-7 Cell Cardiovascular system Cations Cell Proliferation Cell surface Cells Chest Chloride Ion Chlorides Congestive Heart Failure Cytokine Signaling Deposition Development Disease Distal convoluted renal tubule structure Endothelial Cells Event Future Genetic Health Heart Human Hypokalemic alkalosis Hypomagnesemia IL6 gene Inflammation Inflammatory Inherited Interferon Type II Interleukin-1 beta Interleukin-18 Ion Channel Ion Channel Protein Kidney Knowledge Lead Lesion Lipids Mediating Metolazone Molecular Mus Pathway interactions Patients Phenotype Phosphorylation Site Phosphotransferases Production Proteins Recombinant Interleukin-18 Role Secondary to Serum Severities Signal Pathway Signal Transduction Smooth Muscle Myocytes Sodium Chloride Sodium-Restricted Diet Syndrome T-Lymphocyte Testing Tumor Necrosis Factor-alpha Tyrosine Phosphorylation abdominal aorta age related aortic arch atherogenesis cardiovascular risk factor cell motility chemokine cytokine hypertensive heart disease interleukin-18 binding protein interleukin-18 receptor intraperitoneal macrophage novel overexpression receptor symporter uptake

项目摘要

DESCRIPTION (provided by applicant): Na-Cl co-transporter (NCC) is a 125-kDa 12-transmembrane Na+-dependent cation-chloride co-transporter that is primarily expressed in kidney distal convoluted tubules to resorb the filtered load of NaCl in the kidney. In mice, deficiency of NCC causes Gitelman syndrome, an inherited hypokalemic alkalosis with significant hypocalciuria and hypomagnesemia, which are secondary to the deficit in NaCl resorption. Interleukin 18 (IL18) is an inflammatory cytokine that promotes inflammatory cell cytokine/chemokine (e.g., IFN-γ, IL6, MCP-1) production. In atherosclerotic lesions, increased levels of IL18 and its cognate receptor IL18r are detected in macrophages, smooth muscle cells (SMCs), and endothelial cells (ECs). Serums IL18 levels are elevated in patients with CAD, correlate with CAD severity score, are associated with cardiovascular risk factors, and predict future CAD events. In experimental atherosclerosis, overexpression or intraperitoneal administration of IL18 increases atherosclerotic lesion formation and enhances vulnerable plaque phenotypes via an IFN-γ- dependent pathway, while inactivation or genetic deficiency of IL18 slows lesion progression. Our preliminary studies show that IL18 binds to NCC and mediates downstream protein tyrosine phosphorylation in mouse heart ECs. Stimulation of macrophages, T cells, and aortic SMCs and ECs with inflammatory cytokines (e.g., IL18, TNF-α, IL1ß) induces NCC expression. In both humans and mice, normal aortas express negligible NCC and IL18r, but SMCs, ECs, and macrophages in atherosclerotic lesions express high amounts of both NCC and IL18r. In atherosclerosis-prone apolipoprotein E-deficient (Apoe-/-) mice, the absence of either IL18r or NCC does not slow atherosclerosis development significantly. But combined deficiency of IL18r and NCC significantly reduces aortic arch macrophage content and lesion areas, and thoracic-abdominal aorta lipid deposition, along with reduced serum cytokines IFN-γ and IL6. From cultured macrophages, the combined absence of NCC and IL18r impairs IL18-mediated cell signaling, and significantly reduces IL18 cell-surface binding and consequent inflammatory cytokine and chemokine production. These observations lead to our central hypothesis that NCC and IL18r are alternative IL18-binding molecules on inflammatory cells and cardiovascular cells, and that they mediate IL18 activities cooperatively in atherogenesis and possibly in other inflammatory diseases. We propose three aims: 1). to examine whether inhibition, activation, or genetic deficiency of NCC affects atherogenesis; whether NCC activities on inflammatory cells (e.g. macrophages) are essential to atherogenesis; and age-dependent; 2). To examine whether NCC acts like other kinase receptor, NCC activities in salt uptake and IL18 signaling interplay with each other, IL18 has different affinity for NCC and IL18r, and NCC is specific for IL18; and 3). To examine whether NCC mediates IL18 activities in T cells and vascular SMCs and ECs.

描述（由应用程序提供）：Na-Cl共同转运蛋白（NCC）是125 kDa 12-跨膜Na+依赖性的阳离子 - 氯化物共转运蛋白，主要在肾脏远端储量小管中表达，可在肾脏中恢复Nacl的NaCl滤过负载。在小鼠中，NCC的缺乏会导致Gitelman综合征，Gitelman综合征是一种遗传性低下性碱性病，患有明显的低钙尿和低磁性血症，这是NACL分辨率不足的继发性。白介素18（IL18）是一种炎性细胞因子，可促进炎性细胞因子/趋化因子（例如IFN-γ，IL6，MCP-1）产生。在动脉粥样硬化水平中，在巨噬细胞，平滑肌细胞（SMC）和内皮细胞（ECS）中检测到IL18及其同源受体IL18R的水平升高。 CAD患者的血清IL18水平升高，与CAD严重程度评分相关，与心血管危险因素相关，并预测未来的CAD事件。在实验性动脉粥样硬化中，IL18的过表达或腹膜内施用会增加动脉粥样硬化病变的形成，并通过IFN-γ-依赖性途径增强脆弱的斑块表型，而IL18的失活或遗传性抑制作用却缓慢了病变进展。我们的初步研究表明，IL18与NCC结合并介导小鼠心脏EC中的下游蛋白酪氨酸磷酸化。巨噬细胞，T细胞和主动脉SMC和ECS刺激炎症细胞因子（例如IL18，TNF-α，IL1ß）诱导NCC表达。在人类和小鼠中，正常的主动脉都表现出可以忽略的NCC和IL18R，但是动脉粥样硬化病变中的SMC，ECS和巨噬细胞表现出大量的NCC和IL18R。 E缺乏（APOE-/ - ）小鼠，缺乏IL18R或NCC并不能显着降低动脉粥样硬化的发育。但是IL18R和NCC的综合缺乏可显着降低主动脉弓巨噬细胞含量和病变区域，胸腹主动脉脂质沉积，以及减少的血清细胞因子IFN-γ和IL6。从培养的巨噬细胞中，NCC和IL18R的综合缺失损害了IL18介导的细胞信号传导，并显着降低了IL18细胞表面结合，因此炎症细胞因子和趋化因子产生。这些观察结果导致了我们的中心假设，即NCC和IL18R是炎症细胞和心血管细胞上的替代性IL18结合分子，并且它们在动脉粥样硬化中介导IL18活性在其他炎症疾病中可能介导。我们提出三个目标：1）。检查NCC的抑制作用，激活或遗传缺乏是否会影响动脉粥样硬化； NCC对炎性细胞的活性（例如巨噬细胞）是否对于动脉粥样硬化至关重要；和年龄依赖； 2）。为了检查NCC是否像其他激酶受体一样起作用，NCC摄入中的NCC活性和IL18信号相互作用相互竞争，IL18对NCC和IL18R具有不同的亲和力，而NCC对IL18是特定的。和3）。检查NCC是否介导T细胞以及血管SMC和EC中的IL18活性。