The molecular and cellular mechanisms of the STAT3 mutation-mediated pulmonary disorder in Autosomal Dominant Hyper IgE Syndrome (AD-HIES)

常染色体显性高 IgE 综合征 (AD-HIES) STAT3 突变介导的肺部疾病的分子和细胞机制

• 批准号: 10584596
• 负责人: Richard Charles Boucher
• 金额: $ 73.42万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-05 至 2026-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10584596
• 关键词: Adenosine; Air; Alternative Therapies; Atelectasis; Attention; Bacterial Infections; Bone Marrow; CRISPR/Cas technology; Case Study; Cell Culture Techniques; Cell Differentiation process; Cell physiology; Cells; Characteristics; Chlorides; Chronic; Cilia; Clinical Trials; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Defect; Development; Dominant-Negative Mutation; Dynein ATPase; Eczema; Elements; Eosinophilia; Epithelial Cells; Etiology; Exhibits; Exposure to; Extramural Activities; Failure; Frequencies; Gases; Genes; Genetic Transcription; Goals; Hematopoietic Stem Cell Transplantation; High Resolution Computed Tomography; Host Defense; Human; Hydration status; IgE; Image; Imaging Techniques; Impairment; In Vitro; Infection; Infectious Skin Diseases; Inflammation; Inflammatory; Institution; Intramural Research Program; Ions; Job' s Syndrome; Lung; Lung diseases; Lung infections; Magnetic Resonance Imaging; Measurement; Measures; Mediating; Medical center; Metabolic Clearance Rate; Modality; Molecular; Morbidity - disease rate; Motor Activity; Mucociliary Clearance; Mucous Membrane; Mucous body substance; Mutation; Mycoses; National Heart, Lung, and Blood Institute; National Institute of Allergy and Infectious Disease; Nose; Obstruction; Pathogenesis; Pathway interactions; Patients; Pattern; Performance; Phenotype; Physiological; Pneumonia; Primary Ciliary Dyskinesias; Procedures; Property; Rare Diseases; Recurrence; Regulation; Resistance; Risk; Role; STAT3 gene; Saline; Stimulus; Surface; Survivors; System; T-Lymphocyte; Technology; Testing; Therapeutic; Thick; Transgenic Mice; United States National Institutes of Health; Viscosity; Water; X-Ray Computed Tomography; airway epithelium; autosome; bronchial epithelium; chronic infection; cilium motility; clinical center; congenital immunodeficiency; defense response; ectoATPase; epithelial repair; imaging modality; immune reconstitution; improved; in vitro Assay; in vitro activity; in vitro testing; in vivo; inhibitor; injury and repair; loss of function; microCT; mortality; mucus clearance; mutant; novel; novel therapeutics; overexpression; quantitative imaging; rare genetic disorder; reconstitution; respiratory imaging; small molecule; targeted treatment; therapeutic evaluation; transcription factor; ultra high resolution; ventilation

Abstract: Autosomal Dominant Hyper IgE Syndrome (AD-HIES) is a rare genetic disease characterized by elevated IgE, eosinophilia, eczema, recurrent skin infections, and pneumonia. AD-HIES is most frequently caused by mutations in the STAT3 gene, leading to impaired TH17 cell differentiation and recurrent pulmonary infections, a major cause of morbidity and mortality in AD-HIES patients. Airway mucus obtained from AD- HIES patients is abnormally thick (hyperconcentrated), viscous, and adherent. The abnormal mucus properties are associated with chronic inflammation and mucus obstruction, resembling features observed in cystic fibrosis (CF) and primary ciliary dyskinesia (PCD). In CF and PCD, loss of the CFTR or motile cilia, respectively, leads to hyperconcentrated mucus, impaired mucociliary clearance (MCC), and chronic infection, suggesting candidate pathways for the pathogenesis of AD-HIES lung disease. Our preliminary in vitro and in vivo studies suggest both pathways are defective in AD-HIES: (1) CFTR transcription and function are downregulated; and (2) expression of ciliary shaft dyneins are also downregulated. These data led us to test the hypothesis that defective STAT3 perturbs mucus clearance in AD-HIES lungs. Three Aims are prepared to test this hypothesis. Specific Aim 1: STAT3 regulates CFTR-mediated airway surface hydration and mucus concentration. We will measure nasal PD, sweat chloride values in AD-HIES patients in vivo, and CFTR activity in vitro using primary AD-HIES human bronchial epithelial (HBE) cultures at baseline and after exposure to inflammatory stimuli. Therapeutic approaches aimed to improve CFTR expression and function will be assessed in AD-HIES HBE cells. Specific Aim 2: STAT3 is required for ciliated cell genesis, function, and mucociliary transport. We will image motile cilia ultrastructure in freshly collected nasal scrapes by TEM. Ciliary beat frequency and direction, waveform patterns, regulation of ATP/adenosine concentrations, and mucus clearance rate in the AD-HIES HBE cells will be measured. Therapies aimed to restore MCC and mucus hydration will be assessed for their efficacy in AD-HIES HBE cells. Specific Aim 3: AD-HIES is associated with airway mucus plugging and heterogeneous ventilation that can be quantified using advanced CT and MRI imaging techniques. To quantify the muco-obstructive phenotype in AD-HIES patients in vivo, we will perform quantitative imaging analyses by conventional CT, regional ultra- high resolution CT scan, and high-performance low field MRI to measure mucus plug, airway trapping and gas exchange distribution. This collaborative project combines the strengths of the NHLBI, NIAID, NIH Clinical Center, Johns Hopkins CF Center and UNC’s Marsico Lung Institute, to study AD-HIES lung disease. Data derived from this application should identify the molecular and cellular mechanism(s) of STAT3 in regulating MCC-mediated innate host defense, optimal imaging modalities for detecting mucus obstruction in AD-HIES subjects, and novel therapeutic options for restoring mucus clearance in AD-HIES subjects.

【摘要】：常染色体显性高IgE综合征(AD-HIES)是一种罕见的遗传性疾病,其特征是 最常见的是 IgE 升高、嗜酸性粒细胞增多、湿疹、反复皮肤感染和肺炎。 由 STAT3 基因突变引起，导致 TH17 细胞分化受损和复发性肺结核 感染是 AD-HIES 患者发病和死亡的主要原因。 HIES 患者的粘液异常粘稠（高度浓缩）、粘稠且粘连。 与慢性炎症和粘液阻塞有关，在囊性囊肿中观察到重新组装特征 纤维化 (CF) 和原发性纤毛运动障碍 (PCD) 在 CF 和 PCD 中，CFTR 或活动纤毛丧失。 分别导致粘液浓度过高、粘液纤毛清除功能受损 (MCC) 和慢性感染， 我们的初步体外和体内研究表明了 AD-HIES 肺病发病机制的候选途径。 体内研究表明 AD-HIES 中的两条途径均存在缺陷：(1) CFTR 转录和功能存在缺陷 下调；（2）睫状轴动力蛋白的表达也下调。这些数据促使我们进行测试。 STAT3 缺陷会干扰 AD-HIES 肺部粘液清除的假设是“三个目标”。 准备检验这一假设。具体目标 1：STAT3 调节 CFTR 介导的气道表面。 我们将测量 AD-HIES 患者的鼻腔 PD、汗液氯化物值。 基线时使用原代 AD-HIES 人支气管上皮 (HBE) 培养物进行体内和体外 CFTR 活性 以及暴露于炎症刺激后的治疗方法旨在改善 CFTR 表达和 将在 AD-HIES HBE 细胞中评估功能。 具体目标 2：纤毛细胞需要 STAT3。 我们将对新鲜收集的运动纤毛超微结构进行成像。 通过 TEM 观察鼻刮擦的纤毛跳动频率和方向、波形模式、ATP/腺苷的调节 将测量 AD-HIES HBE 细胞中的浓度和粘液清除率。 将评估恢复 MCC 和粘液水合在 AD-HIES HBE 细胞中的功效。 具体目标 3： AD-HIES 与气道粘液堵塞和异质通气有关，可通过 使用先进的 CT 和 MRI 成像技术来量化粘膜阻塞表型。 在 AD-HIES 患者体内，我们将通过常规 CT、局部超 高分辨率 CT 扫描和高性能低场 MRI，用于测量粘液栓、气道滞留和气体 该合作项目结合了 NHLBI、NIAID、NIH 临床的优势。 中心、约翰霍普金斯 CF 中心和北卡罗来纳大学 Marsico 肺研究所，研究 AD-HIES 肺病数据。 源自该申请的结果应确定 STAT3 调节的分子和细胞机制 MCC 介导的先天最佳宿主防御，用于检测 AD-HIES 中粘液阻塞的成像方式 受试者，以及恢复 AD-HIES 受试者粘液清除的新治疗选择。