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）感染是过敏性哮喘（AA）成年人加重的最常见原因。 AA患者的III型干扰素（IFNL）和T细胞对感染的反应失调，病毒障碍 清除。我们发现过敏原暴露于上皮IFN调节因子（IRF）1- IFNL T细胞共抑制剂编程的死亡配体（PDL）-1/B7H1的抗病毒反应和激活表达。 我们的数据暗示锌指E框（ZEB1）转录因子介导这种表观遗传学 通过结合组蛋白甲基转移酶（EZH2）和组蛋白乙酰转移酶（P300/CBP）重新编程。 特定于发起人的背景。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，并通过免疫调节性嗜酸性粒细胞作用维护。这 Zeb1沉默的表观遗传作用IRF1-IFNL响应却上调PDL1抑制CD8 T细胞 通过在特定启动子环境中募集不同组蛋白的乙酰转移酶来激活。我们的目标 为：1。确定Zeb1如何诱导IRF1-IFNL抗病毒的表观遗传沉默的机制 过敏原和嗜酸性免疫调节的途径。我们将检查Zeb1-Ezh2的效果 在正常和AA上皮细胞中IRF1增强子/启动子的表观遗传修饰沉默（HAEC） 通过精确的核跑（Pro-Seq）和染色质免疫沉淀。我们将研究Zeb-的作用 EZH2在嗜酸性粒细胞调节的抑制原代嗜酸性粒细胞共培养中的IFNL反应。这些 人体ICAM1转基因小鼠将在体内探测途径，并在RV时进行CDE重塑 感染。我们希望有缺陷的IRF/IFNL响应将通过ZEB1-EZH2沉默逆转。 2。阐明 Zeb1如何上调粘膜PD-L1表达并确定其对CD8 T的影响 细胞耐受性，RV清除和AHR。 RV诱导的PD-L1表达将在原代HAEC中测量 沉默ZEB1-P300/CBP途径后。 PDL1对CD8-T细胞抑制的影响将在共同体中进行测试 使用原代人T细胞的培养系统。我们将测试PD-L1在HICAM1中RV清除率中的作用 使用阻断抗体（ABS）的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。我们预计PD-L1抑制作用将增强CD8+ T细胞的激活和 块AHR。 3。测试RV16感染对人粘膜IFNL和PD-L1表达的影响 具有或没有过敏原的志愿者会诱导重塑。我们将招募嗜酸性AAS的志愿者， 过敏性鼻炎（AR）和正常对照。我们将测试重塑诱导的IRF1/IFNL之间的关系 响应重组RV16感染的PD-L1表达。我们期望IRF1-IFNL响应 和CD8+T细胞响应将在AAS中钝化，并具有活性重塑。这个项目将大大发展 我们对粘膜免疫的表观遗传控制的理解，并提供了恢复正常的新策略 AA中的粘膜先天防御。