Effects of miR-21 and miR-155 inhibition in SLE

描述(申请人提供)：系统性红斑狼疮(SLE，狼疮)是一种慢性系统性自身免疫性疾病，其特征是产生和存活自身反应抗体，免疫复合物沉积到各种组织，导致器官损害。狼疮主要影响育龄妇女，男女比例为9：1。与普通人群相比，系统性红斑狼疮的死亡率更高。较高的死亡风险与女性、较年轻的年龄、较短的SLE病程和非裔美国人有关。研究表明，系统性红斑狼疮在非裔美国人、西班牙裔和其他少数民族妇女中更为严重。在过去的二十年里，非裔美国女性的狼疮死亡率上升了67.8%。研究结果表明，这可能与非裔美国人患者更糟糕的肾脏受累和预后有关。肾脏和其他严重的系统性红斑狼疮表现对目前的治疗方式反应不佳，经常需要替代治疗。MicroRNAs(MiRNAs)调节过多的正常细胞和发育过程，它们的功能与人类疾病有关。MiRNAs在人类和小鼠SLE中异常表达，但它们在狼疮免疫反应中的具体作用尚不清楚。我们研究了miRNAs在SLE中的作用；我们的一般假设是，几个miRNAs参与了狼疮免疫调节的紊乱。我们的长期目标是鉴定和干扰小鼠和人类SLE中常见的miRNA调节通路，并研究合成的miRNA抑制剂作为狼疮新的治疗方向。我们的初步研究表明，LNA AntimiRs在体内有效地拮抗外周血淋巴细胞内源性miRNAs，LNA miR-21抑制可改善B6.Sle123的自身免疫表现。我们将使用相同的新方法进行体内和体外研究，以表征miR-21和miR-155在狼疮免疫系统中的功能。AIM的体内研究我将介绍miR-21和miR-155在遗传性SLE模型中对自身抗体产生、淋巴细胞功能和严重的终末器官表现的作用。在可诱导模型的研究中，我们将检查针对致病亲本细胞的选择性miRNA抑制的效果。我们将研究miR-21和miR-155可能的协同作用，并确定它们在免疫系统细胞中的靶点。AIM II上的实验将告诉我们 美国关于miR-21和miR-155在狼疮T细胞活化中的作用。同时，我们将采用两种新的高通量方法，即HITS-CLIP和RNA SEQ，克隆和测序小鼠SLE B和T淋巴细胞、树突状细胞和巨噬细胞中所有与生物学相关的miRNAs。据我们所知，HITS-CLIP以前没有在SLE中进行过，HITS-CLIP和RNAseq的联合结果将提供狼疮小鼠免疫系统细胞中由miRNAs直接调控的所有基因的全景图。

DESCRIPTION (provided by applicant): Systemic lupus erythematosus (SLE, lupus) is a chronic systemic autoimmune disease characterized by production and survival of autoreactive antibodies and deposition of immune complexes to various tissues, leading to organ damage. Lupus affects primarily women of reproductive age, with a female to male ratio of 9:1. Mortality in SLE is increased compared to the general population. Higher risk of death is associated with female sex, younger age, shorter SLE duration and African American race. Studies have shown that SLE is more severe among African American, Hispanic and women of other ethnic minorities. Over the last two decades, lupus mortality rates increased by 67.8% among African American women. Results of studies have suggested that this may be related to worse renal involvement and outcome in African American patients. Renal and other severe SLE manifestations respond poorly to current therapeutic modalities and often require replacement therapy. microRNAs (miRNAs) regulate a plethora of normal cellular and developmental processes and their function is linked to human disease. miRNAs are aberrantly expressed in human and mouse SLE, however their specific role in the immune response in lupus is not well understood. We study the function of miRNAs in SLE; our general hypothesis is that several miRNAs contribute to the disordered immunoregulation in lupus. Our long-term objective is to characterize and interfere with miRNA-regulated pathways common in mouse and human SLE and to investigate synthetic miRNA inhibitors as putative novel therapeutic direction in lupus. Our preliminary studies showed that LNA antimiRs efficiently antagonize endogenous miRNAs in peripheral lymphocytes in vivo and that LNA miR-21 inhibition ameliorates autoimmune manifestations in B6.Sle123. We will employ the same novel methodology for our in vivo and in vitro studies, to characterize the function of miR-21 and miR-155 in the immune system in lupus. In vivo studies in Aim I will inform us on role of miR-21 and miR-155 on autoantibody production, lymphocyte function and severe end-organ manifestations on genetic SLE models. In studies of inducible models we will examine the effect of selective miRNA inhibition targeting pathogenic, parental cells. We will investigate putative synergistic effect of miR-21 and miR- 155 and we will identify their targets in cells of the immune system. Experiments in Aim II will inform us on the role of miR-21 and miR-155 on T cell activation in lupus. In parallel, we will clone and sequence biologically relevant targets of all miRNAs in mouse SLE B and T lymphocytes, dendritic cells and macrophages, by employing two novel high throughput methods, HITS-CLIP and RNA seq. To our knowledge HITS-CLIP has not been previously performed in SLE and the combined results of HITS-CLIP and RNAseq will offer a panoramic view of all genes directly regulated by miRNAs in cells of the immune system in mouse lupus.