Asb2 in CD4+ T cell lineage differentiation and its plasticity

Asb2在CD4 T细胞谱系分化及其可塑性中的作用

基本信息

批准号：
8660033
负责人：
Xiao-Hong Sun
金额：
$ 21万
依托单位：
OKLAHOMA MEDICAL RESEARCH FOUNDATION
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-05-01 至 2016-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8660033
关键词：
Animal Model Ankyrin Repeat Antigens Asthma Autoimmunity Biological Assay Bone Marrow Bone Marrow Transplantation Boxing CD4 Positive T Lymphocytes Cell Differentiation process Cell Lineage Cells ChIP-seq Complex Effector Cell Ensure Equilibrium Evaluation Exhibits F-Box Proteins Funding Genes Genetic Recombination Helper-Inducer T-Lymphocyte Immune System Diseases Immunity In Vitro Infection Inflammatory Bowel Diseases Investigation Knockout Mice Laboratories MADH4 gene Maintenance Measures Mediating Modeling Multiple Sclerosis Mus Notch Signaling Pathway Parasites Pattern Play Process Production Proteins Reaction Regulation Regulatory T-Lymphocyte Role Schistosoma mansoni Serum Signal Transduction T cell differentiation T-Lymphocyte T-Lymphocyte Subsets TCF3 gene Testing Th2 Cells Therapeutic Transplantation Ubiquitin Ubiquitination Viral Work aluminum sulfate base cytokine egg fighting genome wide association study in vivo interest mouse model notch protein promoter protein degradation public health relevance response transcription factor ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): The overall objective of this R21 application is to explore the role of ankyrin-repeat SOCS-box containing protein 2 (Asb2) in the regulation of CD4 helper T cell differentiation. Our previous studies have shown that the Asb2 gene is transcriptionally activated by Notch signaling and Asb2 mediates Notch-induced protein ubiquitination through bridging the formation of dimeric E3 ubiquitin ligase complexes. Asb2 promotes the degradation a large number of proteins including some related to T helper cell differentiation. Interestingly, genome-wide studies have revealed that Asb2 is highly expressed in Th2 cells and repressed in Treg cells. The interesting pattern of expression suggests a potential role for Asb2 in helper T cell differentiation. We hypothesize: (1) Asb2 is crucial for T2 differentiation and maintenance of Th2 lineage fate; (2) Asb2 favors T effector cell fates while suppressing Treg differentiation and destabilizing Treg identifies. We have already generated mouse models in which gain or loss of Asb2 function can be achieved via Cre- mediated recombination. These animal models will be used to test these hypotheses in two aims. Aim1 will explore the role of Asb2 in Th2 differentiation and the stability of the Th2 fate. We will use Asb2 knockout mice to test if loss of Asb2 impairs Th2 differentiation in vitro or destabilizes the Th2 fate when switched to polarizing conditions for Th1, Th17 or Treg. We will then evaluate the role of Asb2 in Th2 responses in vivo by immunizing wild type and Asb2-/- mice with a parasite antigen, the soluble egg antigen (SEA) from Schistosoma mansoni or with OVA in alum. Th2 cytokine secretion and Th2-dependent Ig production in immunized mice will be measured. Aim2 will explore the role of Asb2 in suppressing Treg differentiation. We will ectopically express Asb2 from the ROSA26 promoter in na¿ve T cells and test if Asb2 can block iTreg differentiation in vitro or if Asb2 potentiates the conversion of Treg cells into Th1, Th2 and Th17 lineages. We will also examine Treg differentiation in vivo in Asb2-expressing mice and assess the ability of Asb2- expressing bone marrow to rescue the immune disorder caused by Foxp3-deficient scurfy bone marrow transplanted into RAG1 deficient hosts. Finding from these studies will provide an initial evaluation concerning the role of Asb2 in T helper cell differentiation and lay the ground work for a full-blown in-depth investigation. This line of investigation examines the regulation of T helper cell differentiation from a unique perspective, namely ubiquitin-mediated degradation of regulators of the differentiation. Fresh ideas will likely emerge from these studies that will be o therapeutic value in the contest of immunity and autoimmunity.

描述（由应用程序提供）：此R21应用的总体目标是探索含有蛋白2（ASB2）的Ankyrin-Repeat Socs-box在CD4 Helper T细胞分化调节中的作用。我们先前的研究表明，ASB2基因通过Notch信号传导在转录上激活，ASB2通过桥接二聚体E3泛素连接酶复合物的形成来介导Notch诱导的蛋白质泛素化。 ASB2促进降解大量蛋白质，包括与T辅助细胞分化有关的一些蛋白质。有趣的是，全基因组的研究表明，ASB2在Th2细胞中高度表达并在Treg细胞中受到抑制。有趣的表达模式表明ASB2在辅助T细胞分化中的潜在作用。我们假设：（1）ASB2对于T2谱系命运的T2分化和维持至关重要；（2）ASB2有利于T效应细胞的命运，同时抑制Treg分化和破坏Treg鉴定的稳定。我们已经生成了小鼠模型，其中可以通过重组来实现ASB2功能的增益或丧失。这些动物模型将以两个目的来检验这些假设。 AIM1将探索ASB2在Th2分化和TH2命运的稳定性中的作用。我们将使用 ASB2敲除小鼠测试ASB2的损失是否会在体外损害Th2分化或破坏稳定当切换到TH1，TH17或Treg的偏振条件时，Th2命运。然后，我们将通过用寄生虫抗原，Mansoni血吸虫的固体鸡蛋抗原（SEA）或校友中的OVA来评估ASB2在体内Th2反应中的作用。将测量免疫小鼠中的Th2细胞因子分泌和Th2依赖性Ig产生。 AIM2将探索ASB2在抑制Treg分化中的作用。我们将在Na¿VET细胞中的Rosa26启动子中进行生态表达ASB2，并测试ASB2是否可以在体外阻断ITREG分化，或者ASB2是否可能将Treg细胞转化为Th1，Th2和Th17谱系。我们还将检查表达ASB2的小鼠体内的Treg分化，并评估表达ASB2的骨髓的能力，以挽救由FoxP3缺陷型骨髓骨髓引起的免疫疾病，这些骨髓移植到RAG1缺乏宿主中。从这些研究中发现，将提供有关ASB2在T辅助细胞分化中的作用的初步评估，并为进行全面的深入研究奠定了基础工作。这一调查考试的调节从独特的角度，即泛素介导的分化调节剂的降解，T辅助细胞分化。这些研究可能会出现新的想法，这些研究将在免疫和自身免疫竞赛中具有治疗价值。