Epigenetic Mechanism Reprogramming Mucosal Anti-viral Immunity in Allergic Asthma

过敏性哮喘粘膜抗病毒免疫的表观遗传机制重编程

• 批准号: 10553704
• 负责人: Allan R. Brasier
• 金额: $ 68.12万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-10 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10553704
• 关键词: Adult; Affect; Air; Airway Disease; Allergens; Allergic rhinitis; Antiviral Response; Asthma; Binding; Biopsy; Blocking Antibodies; CD8-Positive T-Lymphocytes; Cells; Chronic; Coculture Techniques; Complex; Data; EP300 gene; EZH2 gene; Enhancers; Eosinophilia; Epigenetic Process; Epithelial Cells; Epithelium; Extracellular Matrix; Extrinsic asthma; FDA approved; Felis catus; Healthcare; Homeobox; Host Defense; Human; Human Volunteers; Hypersensitivity; ICAM1 gene; IRF1 gene; Impairment; In Vitro; Individual; Infection; Inflammatory; Inflammatory Response; Injury; Interferon Suppression; Interferons; Kinetics; Knowledge; Leukocytes; Liquid substance; Lymphocyte; Malignant neoplasm of lung; Measures; Mediating; Mediator; Metaplasia; Methyltransferase; Modeling; Modification; Mucosal Immunity; Mucous Membrane; Mucous body substance; Mus; Nose; Nuclear; Organoids; Pathogenesis; Pathway interactions; Patients; Phase; Play; Population; Predisposition; Production; Quality of life; Recombinants; Relapse; Rhinovirus; Rhinovirus infection; Role; Running; Signal Transduction; Submucosa; System; T cell response; T-Cell Activation; T-Lymphocyte; Testing; Time; Transforming Growth Factor beta; Transgenic Mice; Up-Regulation; Validation; Viral; Virus Diseases; Zinc Fingers; airborne allergen; airway hyperresponsiveness; airway remodeling; antiviral immunity; asthmatic; atopy; chemokine; chromatin immunoprecipitation; chronic inflammatory disease; clinically significant; cytokine; dander; eosinophil; epigenetic silencing; histone acetyltransferase; histone methyltransferase; immune function; immunoregulation; in vivo; inhibitor; mouse model; paracrine; pembrolizumab; preservation; programmed cell death ligand 1; promoter; pulmonary function; recruit; research clinical testing; respiratory infection virus; response; small molecule inhibitor; transcription factor; volunteer

PROJECT SUMMARY/ABSTRACT Rhinovirus (RV) infections are the most common causes of exacerbations in adults with allergic asthma (AA). Patients with AA have dysregulated type III interferon (IFNL) and T cell responses to infection, impairing viral clearance. We have found that allergen exposures silence the epithelial IFN regulatory factor (IRF)1- IFNL antiviral response and activate expression of the T cell co-inhibitor programmed death ligand (PDL)-1/B7H1. Our data implicate the Zinc Finger E box (ZEB1) transcription factor in mediating this epigenetic reprogramming by binding histone methyltransferase (EZH2) and histone acetyltransferase (p300/CBP) in a promoter-specific context. We will test the hypothesis that epithelial ZEB1 is induced by TGFβ signals produced by innate-induced remodeling and maintained by immunomodulatory eosinophil action. The epigenetic actions of ZEB1 silence the IRF1-IFNL response yet upregulate PDL1 to suppress CD8 T cell activation by recruitment of distinct histone acetyltransferases in specific promoter contexts. Our aims are to: 1. Determine the mechanism how ZEB1 induces epigenetic silencing of the IRF1-IFNL anti-viral pathway by allergen-and eosinophil-immunomodulation. We will examine the effect of ZEB1-EZH2 silencing on epigenetic modifications of the IRF1 enhancer/promoter in normal and AA epithelial cells (hAECs) by precision nuclear run-on (PRO-Seq) and chromatin immunoprecipitation. We will examine the role of ZEB- EZH2 in eosinophil-modulated suppression of IFNL responses in primary eosinophil co-cultures. These pathways will be probed in vivo by human ICAM1 transgenic mice with or without CDE remodeling upon RV infection. We expect the defective IRF/IFNL response will be reversed with ZEB1-EZH2 silencing. 2. Elucidate the mechanism how ZEB1 upregulates mucosal PD-L1 expression and determine its effects on CD8 T cell tolerance, RV clearance and AHR. RV-induced expression of PD-L1 will be measured in primary hAECs after silencing ZEB1-p300/CBP pathway. The effect of PDL1 on CD8-T cell suppression will be tested in co- culture systems using primary human T cells. We will test the role of PD-L1 in RV clearance in the hICAM1 transgenic mouse model using blocking antibodies (Abs) and validate the upregulation of PD-L1 in bronchial biopsies of AAs vs normal controls. We expect that PD-L1 inhibition will enhance CD8+ T cell activation and block AHR. 3. Test the effects of RV16 infection on mucosal IFNL and PD-L1 expression in human volunteers with or without allergen induced remodeling. We will recruit volunteers with eosinophilic AAs, allergic rhinitis (AR) and normal controls. We will test the relationship between remodeling induced IRF1/IFNL and PD-L1 expression in response to infection with recombinant RV16. We expect that IRF1-IFNL response and CD8+T cell response will be blunted in AAs with active remodeling. This project will significantly advance our understanding of the epigenetic control of mucosal immunity and provide new strategies to restore normal mucosal innate defenses in AA.

项目摘要/摘要 鼻病毒（RV）感染是成人过敏性哮喘（AA）加重的最常见原因。 AA患者对感染的III型干扰素（IFNL）和T细胞应答失调， 间隙我们已经发现，过敏原暴露沉默上皮细胞干扰素调节因子（IRF）1- IFNL 抗病毒应答并激活T细胞共抑制剂程序性死亡配体（PDL）-1/B7H1的表达。 我们的数据暗示锌指E盒（ZEB 1）转录因子在介导这种表观遗传 通过结合组蛋白甲基转移酶（EZH 2）和组蛋白乙酰转移酶（p300/CBP）进行重编程， 启动子特定的上下文。我们将检验上皮细胞ZEB 1是由TGFβ信号诱导的这一假设 由先天性诱导的重塑产生并由免疫调节嗜酸性粒细胞作用维持。的 ZEB 1的表观遗传作用沉默IRF 1-IFNL反应，但上调PDL 1以抑制CD 8 T细胞 通过在特定启动子环境中募集不同的组蛋白乙酰转移酶来激活。我们的目标 1.确定ZEB 1如何诱导IRF 1-IFNL抗病毒药物的表观遗传沉默的机制 通过过敏原和嗜酸性粒细胞免疫调节途径。我们将研究ZEB 1-EZH 2对细胞凋亡的影响。 在正常和AA上皮细胞（hAEC）中IRF 1增强子/启动子的表观遗传修饰的沉默 通过精确的核运行（PRO-Seq）和染色质免疫沉淀。我们将研究ZEB的作用- EZH 2在原代嗜酸性粒细胞共培养物中嗜酸性粒细胞调节的IFNL应答抑制中的作用这些 将通过人ICAM 1转基因小鼠在RV上进行或不进行CDE重塑来体内探索途径 感染我们预期缺陷的IRF/IFNL应答将被ZEB 1-EZH 2沉默逆转。2.阐明 ZEB 1上调粘膜PD-L1表达的机制及其对CD 8 T细胞的影响 细胞耐受性、RV清除率和AHR。将在原代hAEC中测量RV诱导的PD-L1表达 ZEB 1-p300/CBP通路沉默。PDL 1对CD 8-T细胞抑制的作用将在共培养中测试。 使用原代人T细胞的培养系统。我们将在hICAM 1中测试PD-L1在RV清除中的作用。 使用阻断抗体（Abs）建立转基因小鼠模型，并验证支气管中PD-L1的上调 AAs与正常对照的活检。我们预期PD-L1抑制将增强CD 8 + T细胞活化， 阻止AHR。3.检测RV 16感染对人粘膜IFNL和PD-L1表达的影响 有或没有过敏原诱导的重塑的志愿者。我们会招募嗜酸性粒细胞性动脉瘤的志愿者， 变应性鼻炎（AR）和正常对照。我们将检测重构诱导的IRF 1/IFNL之间的关系， 和PD-L1表达。我们预期IRF 1-IFNL应答 而CD 8 +T细胞反应在具有主动重塑的AA中将减弱。该项目将大大推进 我们对粘膜免疫的表观遗传控制的理解，并提供新的策略，以恢复正常的 AA中的粘膜先天防御。