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.

项目概要/摘要 鼻病毒（RV）感染是成人过敏性哮喘（AA）病情恶化的最常见原因。 AA 患者的 III 型干扰素 (IFNL) 和 T 细胞对感染的反应失调，从而损害病毒 我们发现过敏原暴露会使上皮干扰素调节因子 (IRF)1- IFNL 沉默。 抗病毒反应和 T 细胞辅助抑制剂程序性死亡配体 (PDL)-1/B7H1 的激活表达。 我们的数据表明锌指 E 盒 (ZEB1) 转录因子介导了这种表观遗传 通过结合组蛋白甲基转移酶 (EZH2) 和组蛋白乙酰转移酶 (p300/CBP) 进行重编程 我们将测试上皮 ZEB1 由 TGFβ 信号诱导的假设。 由先天诱导的重塑产生并由免疫调节嗜酸性粒细胞作用维持。 ZEB1 的表观遗传作用沉默 IRF1-IFNL 反应，但上调 PDL1 以抑制 CD8 T 细胞 通过在特定启动子环境中招募不同的组蛋白乙酰转移酶来激活我们的目标。 目的是： 1. 确定ZEB1如何诱导IRF1-IFNL抗病毒药物表观遗传沉默的机制 我们将检查 ZEB1-EZH2 的作用。 正常和 AA 上皮细胞 (hAEC) 中 IRF1 增强子/启动子表观遗传修饰的沉默 通过精密核连续测序 (PRO-Seq) 和染色质免疫沉淀，我们将检查 ZEB- 的作用。 EZH2 在原代嗜酸性粒细胞共培养物中调节嗜酸性粒细胞调节的 IFNL 反应抑制。 将通过人类 ICAM1 转基因小鼠在 RV 上进行或不进行 CDE 重塑来体内探测通路 我们预计，ZEB1-EZH2 沉默可逆转有缺陷的 IRF/IFNL 反应。 2. 阐明。 ZEB1上调粘膜PD-L1表达的机制及其对CD8 T的影响 将在原代 hAEC 中测量细胞耐受性、RV 清除率和 RV 诱导的 PD-L1 表达。 沉默ZEB1-p300/CBP通路后，PDL1对CD8-T细胞抑制的影响将在共测试中进行。 我们将测试 PD-L1 在 hICAM1 中 RV 清除中的作用。 使用阻断抗体 (Abs) 的转基因小鼠模型并验证 PD-L1 在支气管中的上调 AA 与正常对照的活检我们预计 PD-L1 抑制将增强 CD8+ T 细胞的激活和 3.测试RV16感染对人粘膜IFNL和PD-L1表达的影响。 有或没有过敏原诱导重塑的志愿者我们将招募具有嗜酸性 AA 的志愿者， 我们将测试重构诱导的 IRF1/IFNL 之间的关系。 和 PD-L1 表达对重组 RV16 感染的反应，我们预计 IRF1-IFNL 反应。 CD8+T 细胞反应将在 AA 中减弱并进行主动重塑，该项目将显着推进。 我们对粘膜免疫的表观遗传控制的理解并提供恢复正常的新策略 AA 中的粘膜先天防御。