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

• 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
• 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

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，世界上有三分之一的人口患有高血压。在过去的十年里，它 已经清楚免疫激活会导致高血压。活性氧 （ROS）已被证明是这种疾病的重要激活剂。异木兰素 (isoLG) 是 ROS 形成的脂肪酸氧化产物。这些分子共价加成 蛋白质的赖氨酸残基。 IsoLG 加合蛋白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。 T 细胞的这个子集是 仅由 MHC-I 内呈现的肽激活，这意味着蛋白酶体在 肽加工。在初步研究中，我发现 isoLG 加合肽显着 富含系统性红斑狼疮（SLE）患者的单核细胞。我建议解决 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。这将通过以下方式完成 利用 SLE 的 B6.SLE123 小鼠模型。将分析 IsoLG 修饰的肽呈递 通过流式细胞术在基线和高盐处理后进行检测。血压将通过以下方式测量 颈动脉遥测。将通过流式细胞术和组织学分析组织炎症 分析。与 isoLG 清除剂分子 2-HOBA 和高血压刺激物共同治疗将 然后进行。我预测用 2-HOBA 治疗 SLE 小鼠将减轻高血压和 组织炎症。在目标 2 中，我将检验肽的蛋白酶体加工这一假设 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 受体小鼠的高血压刺激。我发现树突状细胞与 TBHP 共同处理 蛋白酶体抑制剂 (PI) 减弱表面 isoLG 呈现。我将与野生型小鼠共同治疗 高血压刺激和 PI。 IsoLG 表面蛋白呈现和组织炎症将 测量。我还发现高血压小鼠 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。我们 已培育出 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。 ！