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: 10393987
• 负责人: Richard Charles Boucher
• 金额: $ 74.85万
• 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/10393987
• 关键词: Adenosine; Air; Alternative Therapies; Atelectasis; Attention; Bacterial Infections; Bone Marrow; CRISPR/Cas technology; 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; Infectious Skin Diseases; Inflammation; Inflammatory; Institutes; 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; Reporting; Resistance; Risk; Role; STAT3 gene; Saline; Stimulus; Surface; Survivors; System; T-Lymphocyte; Technology; Testing; Therapeutic; Thick; Transgenic Mice; United States National Institutes of Health; Water; X-Ray Computed Tomography; airway epithelium; 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.

翻译后摘要：常染色体显性遗传高IgE综合征（AD-HIES）是一种罕见的遗传性疾病，其特征在于， IgE升高、嗜酸性粒细胞增多、湿疹、复发性皮肤感染和肺炎。AD-HIES最常见于 由STAT 3基因突变引起，导致TH 17细胞分化受损和复发性肺动脉高压。 感染是AD-HIES患者发病和死亡的主要原因。得自AD的气道粘液- HIES患者异常粘稠（高浓度）、粘稠和粘连。粘液性质异常 与慢性炎症和粘液阻塞有关，类似于在囊性病变中观察到的特征。 纤维化（CF）和原发性纤毛运动障碍（PCD）。在CF和PCD中，CFTR或运动纤毛的丧失， 分别导致高浓度粘液、粘膜纤毛清除（MCC）受损和慢性感染， 提示AD-HIES肺病发病机制的候选途径。我们初步的体外和体内实验 体内研究表明，AD-HIES中的两种途径都有缺陷：（1）CFTR的转录和功能， 下调;和（2）睫状体动力蛋白的表达也下调。这些数据让我们测试 缺陷性STAT 3干扰AD-HIES肺中粘液清除的假设。三个目标是 准备验证这个假设。具体目标1：STAT 3调节CFTR介导的气道表面 水化和粘液浓缩。我们将测量AD-HIES患者的鼻PD、汗液氯化物值 基线时使用原代AD-HIES人支气管上皮（HBE）培养物的体内CFTR活性和体外CFTR活性 以及暴露于炎症刺激后。治疗方法旨在改善CFTR表达， 将在AD-HIES HBE细胞中评估功能。具体目标2：STAT 3是纤毛细胞所必需的 发生、功能和粘液纤毛运输。我们将在新鲜收集的细胞中对运动纤毛的超微结构进行成像， 鼻刮擦TEM。纤毛搏动频率和方向、波形模式、ATP/腺苷调节 将测量AD-HIES HBE细胞中的浓度和粘液清除率。治疗旨在 将评估恢复MCC和粘液水合作用在AD-HIES HBE细胞中的功效。具体目标3： AD-HIES与气道粘液堵塞和不均匀通气相关， 使用先进的CT和MRI成像技术进行定量。量化粘膜阻塞表型 在体内AD-HIES患者中，我们将通过常规CT、局部超声、 高分辨率CT扫描和高性能低场MRI，以测量粘液栓塞、气道阻塞和气体 交换分布这个合作项目结合了NHLBI，NIAID，NIH临床 中心，约翰霍普金斯CF中心和霍普金斯大学的Marsico肺研究所，研究AD-HIES肺病。数据 从本申请中获得的结果应该鉴定STAT 3在调节细胞凋亡中的分子和细胞机制。 MCC介导的先天性宿主防御，检测AD-HIES中粘液阻塞的最佳成像方式 受试者，以及恢复AD-HIES受试者粘液清除的新治疗选择。