Deaf1 isoforms control changes in PTA expression in the NOD PLN during T1D pathog

Deaf1亚型控制T1D病理过程中NOD PLN中PTA表达的变化

基本信息

批准号：
8485528
负责人：
CHARLES GARRISON FATHMAN
金额：
$ 37.27万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-07-01 至 2015-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8485528
关键词：
Address Age Age of Onset Alternative Splicing Antigens Autoantibodies Autoantigens Autoimmune Diseases Autoimmune Process Beta Cell Binding Biological Assay Biological Process Birth Blood Glucose Cells Cervical lymph node group ChIP-on-chip Collaborations Cytoplasm Data Defect Development Disease Disease Progression Dominant-Negative Mutation Down-Regulation Ectopic Expression Effector Cell Gene Expression Genes Genetic Genetic Transcription Histones Hormones Human Immunohistochemistry Immunophenotyping In Situ Hybridization Inbred NOD Mice Incidence Infiltration Inflammation Inflammatory Insulin Insulin-Dependent Diabetes Mellitus Islet Cell Islets of Langerhans Knock-out Knockout Mice Laboratories Lead Length Letters Lymph Node Tissue Lymphocyte Lymphoid Tissue Measures Mediating Microarray Analysis Modeling Molecular Mus Mutation Non obese Onset of illness Pancreas Pathogenesis Pathway interactions Patients Peptides Peripheral Phenotype Play Production Promoter Regions Protein Binding Protein Isoforms RNA Splicing Regulation Regulatory T-Lymphocyte Reporter Reporting Research Personnel Role Self Tolerance Severities Severity of illness Sorting - Cell Movement Spliced Genes Stromal Cells Structure of beta Cell of islet Syndrome T-Cell Development T-Lymphocyte Testing Thymus Gland Time Tissues Transcriptional Regulation Uveitis Variant age related autoreactive T cell base cell type congenic cytokine design diabetic diabetic patient lymph nodes mouse model nuclear factor 1 peripheral tolerance promoter public health relevance type I diabetic uveoretinitis

项目摘要

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) develops from the autoimmune destruction of pancreatic beta cells by self-reactive T cells. Potentially self-reactive T cells escaping elimination in the thymus may subsequently be deleted or tolerized in the periphery. Recently, stromal cells of lymph nodes were reported to control self-tolerance by inducing the ectopic expression of various peripheral tissue antigens (PTAs) and presenting them to T cells in a manner that induces tolerance (deletion or regulation). Currently, both the transcriptional control of PTA expression in lymph nodes and the dysregulation of peripheral tolerance in T1D are unclear. Our laboratory recently identified Deaf1 as a transcriptional regulator of PTA gene expression in peripheral lymph nodes. Our findings show that the gene expression of PTAs, including insulin and other beta cell antigens, are down- regulated in the pancreatic lymph nodes (PLN) of non-obese diabetic (NOD) mice at 12 weeks of age. This change in expression coincides in time with the onset of beta cell destruction by autoreactive T cells. Preliminary studies show that Deaf1 regulates the expression of multiple PTA genes in the PLN. Thus far, two forms of Deaf1, a full-length functional form and a non-functional variant (Deaf1-VAR) have been identified in the PLN of 12 week-old NOD mice. At this age, Deaf1-VAR expression is significantly higher and Deaf1 expression is significantly lower in the PLN of NOD compared to NOD.B10 mice. Deaf1-VAR is expressed in the cytoplasm, has little transcriptional activity of its own, and inhibits the transcriptional activity of Deaf1 by heterodimerizing with it and retaining it in the cytoplasm. Remarkably, we identified an equivalent alternatively spliced Deaf1 isoform in the PLN of T1D patients. This human variant which was ~20-fold more abundant in the PLN of T1D patients than controls, behaves like the mouse Deaf1-VAR. The high expression of this variant also correlated with the absence of insulin gene expression in the PLN of T1D patients. Based on these data, it is hypothesized that Deaf1 controls the expression of and maintains peripheral tolerance to certain pancreatic beta-cell antigens, including insulin, in the PLN, and that a change in Deaf1 splicing contributes to the pathogenesis of T1D. To examine this hypothesis, proposed studies address the following: 1) What cell type in the PLN expresses Deaf1, Deaf1-VAR and PTAs? 2) How does Deaf1 mediate PTA gene transcription on a molecular level? 3) Does inflammation of the PLN or various "genetic factors" regulate the splicing of Deaf1? 4) Will knock-out of Deaf1 expression in PTA-expressing cells exacerbate NOD disease, and if so, is this due to a lack of autoreactive T cell deletion or a lack of regulatory T cell development? Finally, do either mouse models of autoimmune uveoretinitis or human APS 1 patients have changes in Deaf1 that can be correlated with disease? The results of these studies will provide an overall understanding of how Deaf1 and its isoforms maintain peripheral tolerance and thus, how a change in the splicing of Deaf1 or a mutation in DEAF1 may contribute to the development of autoimmune disease.

描述（由申请人提供）：1 型糖尿病 (T1D) 是由自身反应性 T 细胞对胰腺 β 细胞的自身免疫性破坏而发展而来。在胸腺中逃脱消除的潜在自身反应性 T 细胞可能随后在外周被删除或耐受。最近，据报道，淋巴结的基质细胞通过诱导各种外周组织抗原（PTA）的异位表达并以诱导耐受（缺失或调节）的方式将它们呈递给T细胞来控制自身耐受。目前，T1D 淋巴结中 PTA 表达的转录控制和外周耐受的失调尚不清楚。我们的实验室最近发现 Deaf1 是外周淋巴结中 PTA 基因表达的转录调节因子。我们的研究结果表明，12 周龄非肥胖糖尿病 (NOD) 小鼠的胰腺淋巴结 (PLN) 中 PTA（包括胰岛素和其他 β 细胞抗原）的基因表达下调。这种表达变化与自身反应性 T 细胞破坏 β 细胞的时间一致。初步研究表明Deaf1调节PLN中多个PTA基因的表达。迄今为止，已在 12 周龄 NOD 小鼠的 PLN 中鉴定出两种形式的 Deaf1，即全长功能形式和非功能变体 (Deaf1-VAR)。在这个年龄，与NOD.B10小鼠相比，NOD的PLN中Deaf1-VAR表达显着较高，而Deaf1表达显着较低。 Deaf1-VAR在细胞质中表达，其自身几乎没有转录活性，通过与Deaf1异二聚化并将其保留在细胞质中来抑制Deaf1的转录活性。值得注意的是，我们在 T1D 患者的 PLN 中发现了等效的选择性剪接 Deaf1 同工型。这种人类变体在 T1D 患者的 PLN 中含量比对照组高约 20 倍，其行为类似于小鼠 Deaf1-VAR。该变异体的高表达还与 T1D 患者 PLN 中胰岛素基因表达的缺失相关。基于这些数据，推测 Deaf1 控制 PLN 中某些胰腺 β 细胞抗原（包括胰岛素）的表达并维持其外周耐受性，并且 Deaf1 剪接的变化有助于 T1D 的发病机制。为了检验这一假设，拟议的研究涉及以下问题：1）PLN 中的哪种细胞类型表达 Deaf1、Deaf1-VAR 和 PTA？ 2）Deaf1如何在分子水平上介导PTA基因转录？ 3）PLN的炎症或各种“遗传因素”是否调节Deaf1的剪接？ 4) PTA表达细胞中Deaf1表达的敲除是否会加剧NOD疾病，如果是的话，这是由于缺乏自身反应性T细胞缺失或缺乏调节性T细胞发育造成的吗？最后，自身免疫性葡萄膜视网膜炎小鼠模型或人类 APS 1 患者是否存在与疾病相关的 Deaf1 变化？这些研究的结果将提供对 Deaf1 及其亚型如何维持外周耐受性的全面了解，以及 Deaf1 剪接的变化或 DEAF1 突变如何可能导致自身免疫性疾病的发展。