Modeling the premature airway with neonatal airway basal cells

用新生儿气道基底细胞模拟早产儿气道

• 批准号: 10427449
• 负责人: Paul Hubert Lerou
• 金额: $ 22.35万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-11 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10427449
• 关键词: 37 weeks gestation; ATAC-seq; Affect; Age; Air; Asthma; Attention; Automobile Driving; Autopsy; Basal Cell; Biological Models; Birth; Bronchopulmonary Dysplasia; Cell Line; Cell physiology; Cells; Childhood; Chromatin; Cilia; Complement; Data; Defect; Development; Disease; Elderly; Epigenetic Process; Epithelial; Epithelial Cells; Exhibits; Experimental Models; Foundations; Future; Gene Expression; Genes; Genetic Transcription; Gestational Age; Goals; Host Defense; Human; Immune response; Immune system; Immunity; Impairment; Incidence; Infant; Infection; Inflammation; Inflammatory; Inflammatory Response; Inhalation; Kinetics; Life; Link; Liquid substance; Live Birth; Lung; Measures; Memory; Methods; Modeling; Molecular; Molecular Profiling; Morbidity - disease rate; Mucociliary Clearance; Neonatal; Newborn Infant; Pathway interactions; Patients; Phenotype; Predisposition; Pregnancy; Premature Infant; Prevention; Prevention strategy; Production; Recurrence; Respiratory Disease; Respiratory Syncytial Virus Infections; Respiratory Tract Infections; Respiratory syncytial virus; Risk; Risk Factors; STEM field; Severities; Signal Pathway; Signal Transduction; Structure; System; Systems Biology; Testing; Viral Respiratory Tract Infection; adaptive immune response; airway epithelium; cell type; cohort; cytokine; differential expression; experimental study; gene regulatory network; high risk; improved; infancy; infection risk; innovation; interest; lung development; pathogen; premature; preterm newborn; pulmonary function; respiratory; respiratory morbidity; response; response to injury; stem cells; transcriptome; transcriptome sequencing; treatment strategy

PROJECT SUMMARY/ABSTRACT Prematurity affects one in nine live births in the US and is a well-described risk factor for more severe and recurrent respiratory infection during infancy and development of asthma. While it is clear that gestational age at birth and age at infection influence severity of respiratory infection, host immunologic response, and risk of long-term impact on lung development and function, our understanding of the mechanism driving these associations remains poorly understand. Limited human infant autopsy studies have revealed aberrations in the development and function of the airway epithelium, which serves as the first line barrier of defense against inhaled pathogens. Models of newborn airway epithelium to study why premature babies are at higher risk of infection and long-term ramifications, such as asthma, are lacking. We have developed a method of isolating airway basal cells from respiratory secretions of newborns born prematurely and at term. These cells can be expanded long-term and retain the ability to differentiate into pseudo-stratified airway epithelium making them a powerful system to model pre-term and full-term airways. In preliminary experiments, cilia differentiated from basal cells isolated from premature newborns are longer, beat slower, are less dense, and have decreased linear mucociliary flow compared to full-term infant controls. We also found that developmental and immunity related gene expression pathways are differentially expressed in pre-term basal cells. Using our innovative method of isolating and growing epithelial cells derived from patient-specific basal cells we will study how prematurity alters airway epithelium function. We will define the airway basal cell defect due to prematurity by phenotyping pre- and full-term basal cells. Basal cells lines from pre- and full-term infants will be differentiated in air-liquid interface culture and their differentiation potential, epithelial and cilliary function will be comprehensively evaluated. We will also study how prematurity impacts the inflammatory response of differentiated basal cells following RSV infection. Finally, gene co-expression networks and chromatin accessibility will be used to identify the molecular signature associated with the basal cell prematurity defect related to differentiation and RSV-infection response. The successful execution of these aims will establish patient-specific derived airway basal cells as a practical and biologically relevant model system. In addition, our results will provide a better understanding of the molecular mechanisms linking prematurity and early life RSV infection with longer-term morbidities such as asthma, which are important for improving prevention and treatment strategies.

项目摘要/摘要 早产会影响美国的九个活产，是一个更严重和更严重的危险因素 婴儿期和哮喘发育期间的复发性呼吸道感染。虽然很明显妊娠年龄 感染时出生和年龄会影响呼吸道感染的严重程度，宿主免疫反应以及 长期对肺发育和功能的影响，我们对驱动这些机制的理解 协会的理解仍然很差。有限的人类婴儿尸检研究表明 气道上皮的发展和功能，它是防御的第一线障碍 吸入病原体。新生气道上皮的模型，以研究为什么早产婴儿的风险更高 缺乏感染和长期影响，例如哮喘。我们已经开发了一种隔离的方法 来自新生儿的呼吸道分泌物的基底细胞过早出生。这些细胞可以是 长期扩大并保留区分伪分层气道上皮的能力 一个强大的系统，用于建模前和完整的气道。在初步实验中，纤毛与 从过早的新生儿分离的基底细胞更长，打击较慢，密度较小，并且减少了 与完整的婴儿对照组相比，线性粘膜纤毛流动。我们还发现发展和免疫力 相关的基因表达途径在术前基底细胞中差异表达。使用我们的创新 分离和生长源自患者特异性基础细胞的上皮细胞的方法，我们将研究如何 早产会改变气道上皮功能。我们将由于早产性而定义气道基底细胞缺陷 表型前和完整的基底细胞。来自前婴儿和完整婴儿的基底细胞线将分化 在空气界面培养及其分化潜力中，上皮和cilliary功能将是 全面评估。我们还将研究早产如何影响 RSV感染后分化的基底细胞。最后，基因共表达网络和染色质 可访问性将用于识别与基础细胞早产缺陷相关的分子签名 与分化和RSV感染响应有关。这些目标的成功执行将建立 患者特异性的衍生气道基底细胞是一种实用且与生物学相关的模型系统。另外，我们的 结果将更好地了解与早产和早期RSV联系的分子机制 长期病毒（例如哮喘）感染，这对于改善预防和 治疗策略。

项目成果

A Tracheal Aspirate-derived Airway Basal Cell Model Reveals a Proinflammatory Epithelial Defect in Congenital Diaphragmatic Hernia.

气管抽吸物衍生的气道基底细胞模型揭示了先天性膈疝的促炎上皮缺陷。

• DOI: 10.1164/rccm.202205-0953oc
• 发表时间: 2023
• 期刊: American journal of respiratory and critical care medicine
• 影响因子: 24.7
• 作者: Wagner,Richard;Amonkar,GaurangM;Wang,Wei;Shui,JessicaE;Bankoti,Kamakshi;Tse,WaiHei;High,FrancesA;Zalieckas,JillM;Buchmiller,TerryL;Zani,Augusto;Keijzer,Richard;Donahoe,PatriciaK;Lerou,PaulH;Ai,Xingbin
• 通讯作者: Ai,Xingbin

Primary culture of tracheal aspirate-derived human airway basal stem cells.

• DOI: 10.1016/j.xpro.2022.101390
• 发表时间: 2022-06-17
• 期刊: STAR protocols