Functional Analysis of MicroRNAs and Target Genes in Immune Tolerance

MicroRNA 和免疫耐受靶基因的功能分析

基本信息

批准号：
10159204
负责人：
DAVID NEMAZEE
金额：
$ 64.94万
依托单位：
SCRIPPS RESEARCH INSTITUTE, THE
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：
2019-06-05 至 2025-05-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10159204
关键词：
Ablation Anabolism Animals Autoimmune Diseases Autoimmunity B-Lymphocytes Biodiversity Biological Process Bone Marrow Chemicals Code Communication Defect Development Disease Dissection Enzymes Failure GADD45A gene Genes Genetic Glycolipids Graft Rejection Grant Hematopoietic System Immune Tolerance Immune system Immunology In Vitro Inbred MRL lpr Mice Knockout Mice Lymphocyte MicroRNAs Modeling Molecular Molecular Analysis Mouse Strains Mutant Strains Mice Nature Pathway interactions Patients Peripheral Play Process Proteins Research Role Therapeutic Transgenic Organisms Untranslated RNA autoreactive B cell cancer immunotherapy central tolerance design diagnostic biomarker immunoregulation in vivo inhibitor/antagonist lupus-like mouse model novel novel therapeutic intervention peripheral tolerance plasma cell differentiation prevent systemic autoimmunity therapeutic target

项目摘要

Project Summary Autoreactive B cells play critical roles in many autoimmune diseases. Multiple immune tolerance checkpoints exist to remove autoreactive B cells or keep them under control. Defects in these checkpoints constitute the basis for the development of autoimmune diseases. Despite intensive study, our understanding of these checkpoints remains incomplete and fragmentary. MicroRNAs (miRNAs) are a new class of small non-coding RNAs that regulate a large diversity of biological processes. Hundreds of miRNAs are expressed in the immune system. While some miRNAs have been shown to play important roles in lymphocyte development and function, the roles of miRNAs in controlling immune tolerance remain poorly understood. We performed in vivo functional analysis of hundreds of miRNAs in the recently established IgMb-macroself mouse model and identified miR-148a as an important regulator of B cell central tolerance and autoimmunity (Nature Immunology 17:433-40, 2016). Further molecular analysis identified 119 target genes regulated by miR-148a in immature B cells. We examined 4 of these target genes and demonstrated that 3 of them, Gadd45a, Bim and Pten, regulate B cell central tolerance. In this proposal, we will 1) further investigate the cellular and molecular mechanisms underlying miR-148a regulation of immune tolerance and autoimmunity, focusing on its role in controlling various B cell tolerance checkpoints and plasma cell differentiation; 2) evaluate the possibility of treating systemic autoimmunity through miR-148a ablation and inhibition by genetic and chemical approaches, respectively, and 3) perform an in vitro functional screen of the other 115 miR-148a target genes to identify novel regulators of B cell tolerance. Our pilot screen has identified B4galt5 as a positive hit. As B4galt5 is a major enzyme in the glycolipid biosynthesis pathway, we speculate that this pathway plays important roles in immune tolerance. Therefore, we will elucidate the function and mechanism of B4galt5 and the glycolipid biosynthesis pathway in controlling B cell tolerance and autoimmunity.

项目摘要自动反应性B细胞在许多自身免疫性疾病中起关键作用。多种免疫耐受性存在检查点以删除自动反应性B单元格或控制它们。这些检查点中的缺陷构成自身免疫性疾病的发展基础。尽管进行了深入研究，但我们对这些检查点的理解仍然不完整和零散。 microRNA（miRNA）是一个新的一类小型非编码RNA调节大量生物学过程。数百个mirnas 在免疫系统中表示。虽然已经显示一些miRNA在淋巴细胞的发育和功能，miRNA在控制免疫耐受性方面的作用仍然很差理解。我们对最近建立的数百个miRNA进行了体内功能分析 IGMB自cACAROSELF小鼠模型，并确定miR-148a为B细胞中央公差的重要调节剂和自身免疫性（自然免疫学17：433-40，2016）。进一步的分子分析确定了119个目标未成熟B细胞中由miR-148a调控的基因。我们检查了这些靶基因的4个，并证明了其中3个GADD45A，BIM和PTEN调节B细胞中心耐受性。在此提案中，我们将进一步研究miR-148a免疫耐受性调节的细胞和分子机制和自身免疫性，重点关注其在控制各种B细胞耐受性检查点和血浆细胞中的作用分化； 2）评估通过miR-148a消融治疗系统自身免疫的可能性分别通过遗传和化学方法抑制和3）进行体外功能屏幕其他115 miR-148a靶基因鉴定了B细胞耐受性的新型调节剂。我们的飞行员屏幕有将B4GALT5识别为积极的命中。由于b4galt5是糖脂生物合成途径中的主要酶，我们推测该途径在免疫耐受性中起重要作用。因此，我们将阐明 B4GALT5的功能和机制和控制B细胞耐受性的糖脂生物合成途径和自身免疫。