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

描述（由申请人提供）：系统性红斑狼疮（SLE, lupus）是一种慢性全身性自身免疫性疾病，其特征是自身反应性抗体的产生和生存以及免疫复合物沉积到各种组织，导致器官损伤。狼疮主要影响育龄妇女，男女比例为9:1。与一般人群相比，SLE患者的死亡率有所增加。较高的死亡风险与女性、年轻、SLE病程较短和非裔美国人种族有关。研究表明，SLE在非裔美国人、西班牙裔和其他少数民族女性中更为严重。在过去的二十年里，非裔美国妇女的狼疮死亡率上升了67.8%。研究结果表明，这可能与非裔美国患者肾脏受累和预后较差有关。肾脏和其他严重SLE表现对目前的治疗方式反应不佳，通常需要替代治疗。microRNAs （miRNAs）调节大量正常细胞和发育过程，其功能与人类疾病有关。mirna在人和小鼠SLE中异常表达，但其在狼疮免疫反应中的具体作用尚不清楚。我们研究了mirna在SLE中的功能；我们的一般假设是，几种mirna有助于狼疮免疫调节紊乱。我们的长期目标是表征和干扰小鼠和人类SLE中常见的miRNA调控通路，并研究合成miRNA抑制剂作为狼疮推定的新治疗方向。我们的初步研究表明，在体内，LNA抗mir能有效拮抗外周淋巴细胞中的内源性mirna，抑制LNA miR-21可改善B6.Sle123的自身免疫表现。我们将采用相同的新颖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 I的体内研究将告诉我们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终末器官表现的作用。在诱导模型的研究中，我们将研究靶向致病性亲本细胞的选择性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的实验将提供信息

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.