A Role of Isolevuglandins in Essential Hypertension and Systemic Lupus Erythematosus

项目摘要 原发性高血压和系统性红斑狼疮（SLE）是美国的毁灭性条件 老兵据估计，世界上有三分之一的人口患有高血压，尽管有大量的 治疗方案。SLE是一种异质性疾病，其治疗仅限于使用非特异性免疫抑制剂。 全身免疫抑制人们对这些疾病的机制缺乏了解。 异evuglandins（IsoLG）是由活性氧形成的脂肪酸的氧化产物。 这些分子与蛋白质的赖氨酸残基共价加合。加合的蛋白质然后呈现为 自身抗原与T细胞结合，导致免疫细胞活化，并导致高血压和全身性自身免疫。 根据以前发表的研究和初步数据，很明显，原发性高血压和 SLE是由这种异LG加合物形成、加工和免疫细胞活化的过程引发的。我有 发现免疫蛋白酶体亚单位LMP 7在异LG加合物的呈递中的重要作用。 原发性高血压小鼠模型中自身抗原、高血压和主动脉炎症的发生 高血压此外，在SLE小鼠模型中，我还发现用异LG治疗， 清除剂，2-羟基苄胺，减弱高血压和全身自身免疫。最后，我发现， SLE患者亚群在抗原呈递细胞内表现出isoLG蓄积，这提示了一种独特的 这些患者的临床特征和潜在治疗机会。我假设在抗原中 在高血压和SLE的呈递细胞中，isoLG加合物由一个 免疫蛋白酶体依赖性机制与免疫蛋白酶体亚基LMP 7的主要作用有关。 此外，SLE患者表现出isoLG-加合物积累表现出独特的疾病特征。我 具体目标是：（1）确定isoLG-加合物在SLE相关高血压和疾病中的作用 异质性(2)确定免疫蛋白酶体功能在isoLG加合抗原呈递中的作用 和高血压。(3)为了确定LMP 7功能在免疫活化中的作用，使用异LG加合抗原 表现和高血压。为了实现这些目标，我们将招募SLE患者， 外周血单个核细胞将通过流式细胞术研究细胞中是否存在异LG加合物 在抗原呈递细胞的独特群体内积累。异LG加合物水平将与 临床参数，以确定与加合物积累相关的参数。为了研究 我将利用免疫蛋白酶体的三个亚基全面缺陷的小鼠 (TKO小鼠）。我还产生了免疫蛋白酶体的胰凝乳蛋白酶亚基的条件敲除 （LMP 7 fl/fl），其将与CD 11 c-Cre转基因动物杂交以产生抗原呈递细胞特异性 LMP 7缺陷动物。将研究这些动物的高血压和炎症的发展， 建立了两种获得性高血压小鼠模型。研究LMP 7的作用 在SLE中，我建议在SLE的B6.SLE123鼠模型中检查LMP 7抑制剂PR-957的作用。 此外，我将研究树突状细胞特异性LMP 7缺失在化学诱导的SLE模型中的作用。 通过用降植烷处理LMP 7 fl/fl/CD 11 c-Cre。将研究动物是否发生高血压， 免疫细胞扩增、自身抗体产生和肾损伤。总之，这些研究有希望 阐明了原发性高血压和SLE的新机制。此外，他们将提供新颖的 治疗这些病症的治疗机会。

PROJECT SUMMARY Essential hypertension and systemic lupus erythematosus (SLE) are devastating conditions for United States Veterans. An estimated one-third of the world’s population suffers from hypertension despite a large number of treatment options. SLE is a heterogeneous disease the treatment of which is limited to the use of non-specific global immunosuppression. There is a lack of understanding of the mechanisms underlying these conditions. Isolevuglandins (IsoLGs) are oxidation products of fatty acids that form as a result of reactive oxygen species. These molecules adduct covalently to lysine residues of proteins. Adducted proteins are then presented as autoantigens to T-cells resulting in immune cell activation and resultant hypertension and systemic autoimmunity. Based upon previously published studies and preliminary data, it is clear that both essential hypertension and SLE are initiated by this process of isoLG-adduct formation, processing, and immune cell activation. I have discovered an important role of the immunoproteasome subunit LMP7 in the presentation of isoLG-adducted autoantigens, the development of hypertension, and aortic inflammation in a mouse model of essential hypertension. Additionally, in a mouse model of SLE, I have also discovered that treatment with an isoLG scavenger, 2-hydroxybenzylamine, attenuates hypertension and systemic autoimmunity. Finally, I found that a subset of patients with SLE exhibit isoLG accumulation within antigen presenting cells, suggesting a unique clinical profile and potential therapeutic opportunities for these patients. I hypothesize that within antigen presenting cells in both hypertension and SLE, isoLG adducts are processed and displayed by an immunoproteasome dependent mechanism with a primary role of the immunoproteasome subunit LMP7. Additionally, patients with SLE that exhibit isoLG-adduct accumulation exhibit unique disease characteristics. My specific aims are: (1) To determine a role of isoLG-adducts in SLE-associated hypertension and disease heterogeneity. (2) To determine the role of immunoproteasome function in isoLG-adducted antigen presentation and hypertension. (3) To determine the role of LMP7 function in immune activation, isoLG-adducted antigen presentation, and hypertension in SLE. To accomplish these aims we will recruit SLE patients and obtain peripheral blood mononuclear cells. Cells will be studied by flow cytometry for the presence of isoLG-adduct accumulation within unique populations of antigen presenting cells. IsoLG-adduct levels will be compared with clinical parameters to determine the parameters that correlate with adduct accumulation. To study the function of the immunoproteasome, I will utilize mice globally deficient for the three subunits of the immunoproteasome (TKO mice). I have also generated a conditional knockout of the chymotrypsin subunit of the immunoproteasome (LMP7fl/fl) which will be crossed to CD11c-Cre transgenic animals to generate an antigen presenting cell specific LMP7 deficient animal. These animals will be studied for the development of hypertension and inflammation in the setting of two well established acquired models of essential hypertension in mice. To study the role of LMP7 in SLE, I propose to examine the effect of the LMP7 inhibitor PR-957 in the B6.SLE123 murine model of SLE. Additionally, I will study the effect of dendritic cell specific LMP7 deletion in a model of chemically-induced SLE by treating LMP7fl/fl/CD11c-Cre with pristane. Animals will be studied for the development of hypertension, immune cell expansion, autoantibody production, and renal injury. Together, these studies hold the promise of elucidating novel mechanistic insights into essential hypertension and SLE. Moreover, they will provide novel therapeutic opportunities for the treatment of these conditions.