Human Airway Cells and Assays

项目摘要/摘要 人类呼吸道细胞和检测核心提供了分化良好的原代培养的体外模型。 人呼吸道上皮细胞用于翻译的、临床前的小分子和靶蛋白检测 以判断它们在提高WT和突变CFRR的密度和/或活性方面的有效性。这些 服务机构在促进对CF发病机制和治疗的翻译评估方面有着出色的记录， 他们还资助了许多在CF研究中心内外资助的研究项目。这个 推动这些努力的总体假设是，由于CFTR缺失或缺陷而导致的离子转运受损 损害呼吸道表面液体(ASL)的体积和组成，阻碍粘液纤毛清除， 并导致感染、炎症和支气管扩张。核心A专注于极化的产生 培养人支气管(HBE)和鼻腔(HNE)上皮细胞，分别取自CF、非CF和非病变肺。 极化条件下通道功能、生物发生和药物介导的突变蛋白校正的研究 HBe细胞，在治疗策略的流水线中预测临床结果。此资源支持 一大批资助项目。 新的方法显著提高了核心产生气液界面(ALI)培养的能力，因此 HBE的可用性不再是速率限制。患者的基因分型与CFTR功能和药物相关 引起离子传输的变化。鼻细胞的一种更直接的方法允许对 更广泛的一组更罕见的基因型别，如CFTR2项目所体现。这些型号由第30页使用 研究人员试图评估CFTR的生物发生，CFTR和ENaC通道的调节，其他 相关转运体、调节ASL体积和组成的机制、炎症机制 由呼吸道感染引起的，以及纠正CF缺陷的治疗。 为了支持对CF发病机制和临床前治疗的机械性评估，Core执行ASL 与中心成员合作进行成分、运输和贩运分析。HBE与USB接口 HNE培养与生理测定优化使用原代呼吸道细胞由中心研究人员和 用于外部学术和行业互动。这个核心提供了细胞、分化的HBE和分析 用于大量新疗法的研究。以离子传输为核心缺陷的CF气道，成功地 极化HBE中的通道生物发生、稳定性和功能的药理学操作 细胞，建立了将治疗策略推进到临床试验所需的翻译疗效数据集。

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.