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-HIE）是一种罕见的遗传疾病，其特征是 升高的IgE，嗜酸性，湿疹，复发性皮肤感染和肺炎。广告最常是 由STAT3基因突变引起，导致TH17细胞分化和复发性肺部受损 感染是广告患者发病率和死亡率的主要原因。从ad-获得的气道粘液 HIES患者的绝对厚度（过度浓度），粘性和粘附。异常粘液特性 与慢性炎症和粘液阻塞有关，类似于囊性的特征 纤维化（CF）和原发性睫状运动障碍（PCD）。在CF和PCD中，CFTR或Mother Cilia的丢失， 导致过度浓缩的粘液，受损的粘膜清除（MCC）和慢性感染， 提示候选肺部肺病发病机理的候选途径。我们的初步体外和 体内研究表明，这两种途径在广告中都是有缺陷的：（1）CFTR转录和功能是 下调； （2）睫状轴动力蛋白的表达也被下调。这些数据导致我们进行测试 有缺陷的STAT3渗透粘液肺中的粘液清除率的假设。三个目标是 准备检验这一假设。特定目标1：STAT3调节CFTR介导的气道表面 水合和粘液浓度。我们将测量广告患者的鼻PD，汗水氯化物值 体内和CFTR活性在基线时使用原发性ad-hies人支气管上皮（HBE）培养 并在暴露于炎症刺激后。旨在改善CFTR表达和的治疗方法 功能将在Ad-Hies HBE细胞中进行评估。纤毛细胞需要特定的目标2：STAT3 创世记，功能和粘膜屈服。我们将在新鲜收集的 鼻腔刮擦。睫状节拍频率和方向，波形模式，ATP/腺苷的调节 将测量AD-HIE HBE细胞中的浓度和粘液清除率。旨在进行治疗 恢复MCC和粘液水合将根据其在AD-HIES HBE细胞中的效率进行评估。具体目标3： 广告烟与气道粘液插头和异质通风有关 使用高级CT和MRI成像技术进行量化。量化粘液 - 刺激性表型 在体内广告患者中，我们将通过常规CT进行定量成像分析 高分辨率CT扫描和高性能低场MRI，以测量粘液插头，气道诱捕和气体 交换分配。这个协作项目结合了NHLBI，NIAID，NIH临床的优势 中心，约翰·霍普金斯（Johns Hopkins）CF中心和UNC的马西科肺研究所（Marsico Lung Institute），研究广告肺疾病。数据 从该应用中得出的应确定STAT3的分子和细胞机制 MCC介导的先天宿主防御，最佳成像方式，用于检测广告中的粘液阻塞 受试者和新型热选择，用于恢复广告主体中的粘液清除率。