Isolevuglandin peptide modification and proteasomal processing is responsible for autoimmune mediated hypertension

异乌兰素肽修饰和蛋白酶体加工是导致自身免疫介导的高血压的原因

基本信息

批准号：
9759492
负责人：
David Patrick
金额：
$ 7万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-22 至 2020-07-21
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9759492
关键词：
Academic Medical Centers Achievement Address Affect Aftercare Age Animal Model Antigen-Presenting Cells Antigens Attenuated Autoantigens Autoimmune Diseases Autoimmune Process Autoimmune Responses Autoimmunity Automobile Driving Award B Cell Proliferation Blood Pressure Blood Vessels CD8-Positive T-Lymphocytes Cardiovascular Diseases Cardiovascular system Carotid Arteries Characteristics Clinical Complex Complication Dendritic Cells Disease Doctor of Philosophy Fatty Acids Flow Cytometry Goals Histologic Hypertension Inflammation Inflammation Process Inflammatory Kidney Knockout Mice Laboratories Lead Lysine Maintenance Measures Mediating Membrane Proteins Modification Morbidity - disease rate Mus Organ Oxidants Oxidative Stress Pathogenesis Patient Care Patients Peptides Physiological Population Post-Translational Protein Processing Prevention Process Proteasome Inhibitor Proteins Reactive Oxygen Species Research Personnel Role Scientist Sodium Chloride Source Stimulus Surface Systemic Lupus Erythematosus T-Cell Activation T-Lymphocyte Telemetry Testing Tissues Training Trees Wild Type Mouse adduct career covalent bond design immune activation immunogenic monocyte mouse model multicatalytic endopeptidase complex novel therapeutics oxidation prevent sex tert-Butylhydroperoxide vascular inflammation

项目摘要

PROJECT SUMMARY An estimated one-third of the world’s population suffers from hypertension. Over the last decade, it has become clear that immune activation contributes to hypertension. Reactive oxygen species (ROS) have been shown to be an important activator of this disease. Isolevuglandins (isoLG) are oxidation products of fatty acids that form as a result of ROS. These molecules adduct covalently to lysine residues of proteins. IsoLG adducted proteins are presented by antigen presenting dendritic cells (DCs) and result in CD8+ T-cell activation and resultant hypertension. This subset of T-cells is exclusively activated by peptides presented within MHC-I, implying a role of the proteasome in peptide processing. In preliminary studies I have found that isoLG-adducted peptides are markedly enriched in monocytes of patients with systemic lupus erythematosus (SLE). I propose to address the hypothesis that IsoLG modification of proteins is responsible for hypertension in SLE and that the presentation of IsoLG-modified peptides is proteasome dependent. In Aim 1, I will test the hypothesis that IsoLG modification of native proteins and their presentation on dendritic cells contribute to hypertension and vascular inflammation in a mouse model of SLE. This will be accomplished by utilizing the B6.SLE123 mouse model of SLE. IsoLG modified peptide presentation will be analyzed by flow cytometry at baseline, and after treatment with high salt. Blood pressure will be measured by carotid artery telemetry. Tissue inflammation will be analyzed by flow cytometry and histologic analysis. Co-treatment with an isoLG scavenger molecule 2-HOBA and hypertensive stimulus will then be performed. I predict that treatment of SLE mice with 2-HOBA will attenuate hypertension and tissue inflammation. In Aim 2, I will test the hypothesis that proteasomal processing of peptides mediates hypertension and isoLG-adduct presentation.. It is known that treatment of mouse dendritic cells with the oxidant tert-butyl hydroperoxide (TBHP) confers isoLG-mediated sensitivity to hypertensive stimulus in recipient mice. I have found that co-treatment of dendritic cells with TBHP and proteasome inhibitor (PI) attenuates surface isoLG presentation. I will co-treat wild-type mice with hypertensive stimulus and a PI. IsoLG surface protein presentation and tissue inflammation will be measured. I have also shown that proteasomal activity is increased in DC’s from hypertensive mice. This correlates with increased expression of the inflammatory immunoproteasomal subunit LMP7. We have generated an LMP7 conditional knockout mouse and will study the effects of LMP7 deletion on hypertension and isoLG-adduct presentation in DCs. !

项目概要据估计，在过去十年中，世界上有三分之一的人口患有高血压。已经清楚免疫活性有助于激活高血压。（ROS）已被证明是这种疾病的重要激活剂。 ROS 形成的脂肪酸氧化产物，这些分子通过共价键加成。蛋白质的赖氨酸残基由抗原呈递树突呈递。细胞 (DC) 并导致 CD8+ T 细胞激活并导致高血压。被 MHC-I 内呈递的肽激活，这意味着蛋白酶体在在初步研究中，我发现 isoLG 加合肽显着增加。我建议解决系统性红斑狼疮（SLE）患者的单核细胞富集问题。假设蛋白质的 IsoLG 修饰是导致 SLE 高血压的原因，并且 IsoLG 修饰肽的呈递依赖于蛋白酶体。在目标 1 中，我将检验该假设。天然蛋白质的 IsoLG 修饰及其在树突状细胞上的呈递有助于 SLE 小鼠模型中的高血压和血管炎症这将通过以下方式实现。将利用 B6.SLE123 小鼠模型对 IsoLG 修饰的肽呈递进行分析。通过流式细胞术测量基线，并在高盐治疗后测量血压。将通过流式细胞术和组织学分析颈动脉遥测。分析显示，用 isoLG 清除剂分子 2-HOBA 和高血压刺激物共同治疗会。我预测用 2-HOBA 治疗 SLE 小鼠将减轻高血压和在目标 2 中，我将检验肽的蛋白酶体加工的假设。介导高血压和 isoLG 加合物呈递。众所周知，小鼠树突状细胞的治疗具有氧化剂叔丁基过氧化氢 (TBHP) 的细胞赋予 isoLG 介导的敏感性我发现树突状细胞与 TBHP 共同治疗受体小鼠的高血压刺激。蛋白酶体抑制剂 (PI) 会减弱表面 isoLG 的表达，我将与野生型小鼠共同治疗。高血压刺激和 PI 表面蛋白的呈现和组织炎症。我还发现，高血压小鼠的 DC 中的蛋白酶体活性有所增加。这与炎症免疫蛋白酶体亚基 LMP7 的表达增加相关。已经培育出 LMP7 条件敲除小鼠，并将研究 LMP7 缺失对高血压和 DC 中 isoLG 加合物的表现。！