Human Airway Cells and Assays

人类气道细胞和检测

• 批准号: 8875228
• 负责人: RAYMOND A FRIZZELL
• 金额: $ 43.48万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-15 至 2018-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8875228
• 关键词: Adenoviruses; Air; Anions; Apical; Archives; Automobile Driving; Bicarbonates; Biogenesis; Biological Assay; Bronchiectasis; Carrier Proteins; Cell Surface Proteins; Cell surface; Cells; Cellular Stress; Cellular biology; Clinical; Clinical Trials; Clinical assessments; Collaborations; Cystic Fibrosis; Cystic Fibrosis Transmembrane Conductance Regulator; Data Set; Defect; Detection; Development; Disease; Disease Progression; Epithelium; Evaluation; Freezing; Funding; Gene Expression; Genotype; Goals; Gold; Human; In Vitro; Infection; Inflammation; Inflammatory; Ion Channel; Ion Transport; Laboratories; Lentivirus Vector; Liquid substance; Lung; Measurement; Mediating; Membrane Protein Traffic; Methods; Modeling; Monitor; Mucins; Mucociliary Clearance; Mucous body substance; Mutation; Nose; Outcome; Pathogenesis; Patients; Permeability; Pharmaceutical Preparations; Phenotype; Physiological; Process; Production; Proteins; Recycling; Regulation; Reporter; Reporting; Research; Research Personnel; Research Project Grants; Resources; Services; Small Interfering RNA; Therapeutic; Time; Tissues; Transduction Gene; Transfection; VX-770; Viral; Water; Work; abstracting; airway epithelium; airway surface liquid; cystic fibrosis airway; density; epithelial Na+ channel; human DICER1 protein; improved; in vitro Model; member; mutant; novel; novel therapeutics; particle; pre-clinical; preclinical study; protein expression; ranpirnase; repository; sample fixation; small hairpin RNA; small molecule; therapeutic evaluation; therapeutic target; trafficking; translational study

Project Summary/Abstract The Human Airway Cell and Assays Core provides in vitro models of well-differentiated, primary cultures of human airway epithelia for translational, pre-clinical assays of small molecules and target protein manipulations to judge their efficacy for improving the density and/or activity of WT and mutant CFTRs. These services have an outstanding record of promoting translational assessments of CF pathogenesis and therapy, and they support numerous funded research projects both within and outside the CF Research Center. The overall hypothesis driving these efforts is that impaired ion transport due to missing or defective CFTR compromises the volume and composition of the airway surface liquid (ASL), impeding mucociliary clearance, and leading to infection, inflammation and bronchiectasis. Core A is focused on the production of polarized cultures of human bronchial (HBE) and nasal (HNE) epithelia from CF, non-CF and non-diseased lungs. Studies of channel function, biogenesis and drug-mediated mutant protein correction, performed in polarized HBE cells, are predictive of clinical outcome in the pipeline of therapeutic strategies. This resource supports a large number of funded projects. New methods have markedly increased the Core's capacity to generate air-liquid interface (ALI) cultures, so that HBE availability is no longer rate-limiting. Patient genotype is correlated with CFTR function and drug- induced changes in ion transport. A more directed approach with nasal cells allows for similar analyses on a broader set of rarer genotypes, as embodied in the CFTR2 project. These models are used by P30 investigators who seek to evaluate CFTR biogenesis, the regulation of CFTR and ENaC channels, other relevant transporters, the mechanisms that regulate ASL volume and composition, inflammatory mechanisms arising from airway infections, and treatments to correct CF defects. To support mechanistic evaluations of CF pathogenesis and pre-clinical therapeutics, the Core performs ASL composition, transport and trafficking assays in collaboration with Center members. The interface of HBE and HNE cultures with physiological assays optimizes the use of primary airway cells by Center investigators and for outside academic and industrial interactions. This Core has provided cells, differentiated HBE and assays for numerous studies of new therapeutics. With ion transport as the core defect in CF airways, the successful pharmacological manipulation of channel biogenesis, stability and function, when performed in polarized HBE cells, establishes the translational efficacy dataset needed for advancing a therapeutic strategy to clinical trials.

项目总结/摘要 人气道细胞和测定核心提供了良好分化的原代培养物的体外模型， 用于小分子和靶蛋白的翻译、临床前测定的人气道上皮 这些方法可以通过对细胞进行操作来判断它们改善WT和突变CFTR的密度和/或活性的功效。这些 在促进CF发病机制和治疗的转化评估方面， 他们支持CF研究中心内外的许多资助研究项目。的 推动这些努力的总体假设是，由于CFTR缺失或缺陷， 损害气道表面液体（ASL）的体积和组成，阻碍粘膜纤毛清除， 并导致感染、炎症和支气管扩张。核心A是专注于生产偏光 来自CF、非CF和非患病肺的人支气管（HBE）和鼻（HNE）上皮细胞的培养物。 研究通道功能，生物起源和药物介导的突变蛋白质校正，在极化 HBE细胞是治疗策略管道中的临床结果的预测。此资源支持 大量资助项目。 新方法显著提高了Core产生气液界面（ALI）培养物的能力， HBE的可用性不再是费率限制。患者基因型与CFTR功能和药物相关。 引起离子运输的变化。一种更直接的鼻细胞方法允许对一种 更广泛的一组罕见的基因型，体现在CFTR2项目。这些型号用于P30 寻求评估CFTR生物发生，CFTR和ENaC通道调节，其他 相关转运蛋白，调节ASL体积和组成的机制，炎症机制 引起的气道感染，以及纠正CF缺陷的治疗。 为了支持CF发病机制和临床前治疗的机制评价，核心进行ASL 与中心成员合作进行成分、运输和贩运分析。HBE与 HNE培养与生理学检测优化了中心研究者对原代气道细胞的使用， 学术界和工业界的互动该核心提供了细胞，分化HBE和分析 用于大量新疗法的研究。以离子转运为CF气道的核心缺陷，成功地 当在极化HBE中进行时，对通道生物发生、稳定性和功能的药理学操纵 细胞，建立了将治疗策略推进到临床试验所需的转化疗效数据集。