The immunoregulatory role of Alveolar Macrophages in Chronic Beryllium Disease

肺泡巨噬细胞在慢性铍病中的免疫调节作用

• 批准号: 9176462
• 负责人: Li Li
• 金额: $ 36.29万
• 依托单位: NATIONAL JEWISH HEALTH
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-30 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9176462
• 关键词: Adenoviruses; Alveolar Macrophages; Antibodies; Berylliosis; Beryllium; Binding; Biological Markers; Breathing; Bronchoalveolar Lavage; CD4 Positive T Lymphocytes; CD55 Antigens; CD80 gene; Cell surface; Cells; Cessation of life; Chronic Disease; Chronic berylliosis; Clinical; Complement; Data; Development; Diagnosis; Disease; Disease Progression; Down-Regulation; FCGR3B gene; Family; Fibrosis; Figs - dietary; Gene Expression; Genes; Goals; Granulomatous; HLA-DP Antigens; HLA-DPB1 gene; HLA-DR Antigens; Immune; Immune Response Genes; Immune response; In Vitro; Individual; JAK1 gene; JAK2 gene; Knowledge; Lead; Light; Longitudinal Studies; Lung; Lung Inflammation; Lung diseases; Mediating; Mediator of activation protein; Modeling; Molecular; Organ; Pathway interactions; Patients; Phenotype; Phosphorylation; Population; Production; Proteins; Public Health; Regulator Genes; Regulatory Pathway; Research; Respiratory Insufficiency; Role; STAT1 gene; STAT3 gene; Sampling; Staging; System; T cell response; TNF gene; TNFRSF5 gene; Testing; Work; Workplace; abstracting; base; crosslink; follow-up; genetic linkage; improved; inhibitor/antagonist; innovation; knock-down; member; neutralizing antibody; novel; novel marker; outcome forecast; overexpression; potential biomarker; prevent; targeted treatment; therapeutic target; transcriptome sequencing

Project Summary/Abstract: The goal of this study is to determine the impact of CD55 and JAK/STAT pathway dysregulation in alveolar macrophages (AMs) on chronic beryllium disease (CBD) progression. CBD is an important organ-specific immune-mediated disease, characterized by granulomatous lung inflammation, fibrosis, and death, due to end-stage respiratory insufficiency. Thus, CBD remains an important public health concern. The precursor to this disease (beryllium sensitization; BeS) progresses to CBD at a rate of approximately 6-8% per year. Although the binding and presentation of Be via βGlu69-containing HLA-DP molecules to pathogenic CD4+ T cells provides an explanation for the genetic linkage of DP2 to CBD and BeS, this does not explain the progression of BeS to CBD. Furthermore, DP2 is found in up to 40% of Be-workers without evidence of BeS or CBD. Thus, additional mechanisms must be involved in progression of BeS to CBD. Our hypothesis is that CBD AMs downregulate the negative immunoregulatory gene CD55 and its pathway and overexpress the positive regulatory activating JAK/STAT pathway, which augments the immune response to Be and progression from BeS to CBD. In Aim 1, we will define if CD55 on AMs is a negative regulator in the immune response to Be in AMs from BeS patients. Specifically, using either the overexpression of the CD55 genes using a adenovirus overexpression system and agonistic antibody or the reduction of CD55 genes using adenovirus knockdown system and neutralizing antibody of CD55, we will determine the impact of alteration of CD55 on consequential functional changes in TNF-α production, phenotype (CD16, CD40, CD80, CD86, HLA- DR), JAK/STAT activation and BeLPT comparing CBD, BeS and controls. Finally, using RNA-seq, we will investigate the regulatory networks associated with the downregulation of CD55 in CBD compared to BeS; In Aim 2, we will determine if the JAK/STAT pathway is a positive regulator of the immune response to Be in AMs and is increased in AMs from subjects with CBD compared to BeS. Specifically, AMs from CBD will be compared to those from BeS and controls subjects, assessing either the overexpression of the JAK/STAT pathway genes using a adenovirus overexpression system or the reduction of the JAK/STAT pathway genes using adenovirus knockdown system and pharmacological JAK1- 3 inhibitors on the Be immune response. We will evaluate the changes in TNF-α production, phenotype (CD16, CD40, CD80, CD86, HLA-DR), and BeLPT. Finally, using RNA-seq we will define the regulatory networks associated with the overactivation of the JAK/STAT pathway. In Aim 3, we will investigate whether the CD55 and JAK /STAT pathway are associated with CBD and progression from BeS to CBD, compared to BeS, using a longitudinal study and validate these findings in a larger longitudinal population to determine if these genes and pathways reveal potential novel biomarkers of diagnosis and prognosis for CBD. At the end of this project we will determine the key regulatory genes and pathways of exposure-mediated immune dysregulation in AMs that are associated with disease progression and reveal potential biomarkers for clinical prognosis and diagnosis. The results obtained from this study will improve our understanding of factors involved in the development of CBD, as well as targets for therapy, and will serve as a model of other exposure-related immune responses and environmentally-induced chronic diseases.

项目摘要/摘要： 这项研究的目的是确定CD55和JAK/STAT途径失调对 肺泡巨噬细胞（AMS）在慢性铍疾病（CBD）进展中。 CBD很重要 器官特异性免疫介导的疾病，其特征是颗粒状肺注射，纤维化和 死亡，由于终末期呼吸不足。这是CBD仍然是一个重要的公共卫生问题。 这种疾病的先驱（铍敏化； BES）以大约6-8％的速度发展为CBD 尽管含有βGlu69的HLA-DP分子与致病性的结合和表现 CD4+ T细胞提供了DP2与CBD的遗传联系的解释，这并不能解释 CBD的进展。此外，在没有BES或 CBD。这，必须参与CBD的BES进展。我们的假设是 CBD AM下调阴性免疫调节基因CD55及其途径和过表达 阳性调节激活JAK/STAT途径，该途径增强了免疫反应和 从BES到CBD的进展。在AIM 1中，我们将定义AMS上的CD55是否是负调节器 免疫反应来自BES患者的AMS。具体而言，使用CD55的过表达 使用腺病毒过表达系统和激动抗体或使用CD55基因还原的基因 腺病毒敲低系统和中和CD55的抗体，我们将确定改变的影响 CD55在TNF-α产生，表型（CD16，CD40，CD80，CD86，HLA- DR），JAK/Stat激活和BELPT比较CBD，BES和对照。最后，使用RNA-Seq，我们将 与BES相比，研究与CBD中CD55下调相关的调节网络；在 AIM 2，我们将确定JAK/STAT途径是否是AMS中免疫响应的积极调节剂 与BES相比，CBD受试者的AM中有所增加。具体来说，来自CBD的AM将 与BES和对照对象相比，评估JAK/STAT的过表达 使用腺病毒过表达系统或JAK/STAT途径降低的途径基因 使用腺病毒敲低系统和Parmaceutical Jak1-3抑制剂在免疫反应上。我们 将评估TNF-α产生，表型（CD16，CD40，CD80，CD86，HLA-DR）和BELPT的变化。 最后，使用RNA-seq我们将定义与过度激活相关的调节网络 jak/stat途径。在AIM 3中，我们将调查CD55和JAK /STAT途径是否关联 与BES相比，使用CBD和从BES到CBD的进展，使用纵向研究并验证这些研究 在较大的纵向种群中的发现，以确定这些基因和途径是否揭示了潜在的新颖 CBD诊断和预后的生物标志物。在该项目结束时，我们将确定关键监管 与疾病相关的AM中暴露介导的免疫失调的基因和途径 进展并揭示了临床预后和诊断的潜在生物标志物。从此获得的结果 研究将提高我们对CBD发展的因素的理解，以及 治疗，并将作为其他与暴露有关的免疫反应和环境引起的模型 慢性疾病。