Quantitative analysis of mucociliary clearance in airway ciliopathies

项目总结 粘液纤毛清除(MCC)是人体呼吸系统重要的机械防御机制。 MCC差是许多遗传性和获得性呼吸道疾病的基本特征，包括原发性 纤毛运动障碍(PCD)、哮喘、慢性支气管炎和囊性纤维化(CF)。由于组织结构复杂 对于肺部，睫状结构的缺陷如何改变功能性微循环，以及局部如何改变，在很大程度上是未知的。 呼吸道上皮结构的变异性，包括细胞类型比例和纤毛搏动参数，影响局部 MCC病理生理学。这些知识缺口极大地削弱了我们预测肺损伤程度的能力。 特殊的睫状体缺陷所致的功能障碍，并了解观察到的呼吸道区域特异性发病 很多肺部疾病。而MCC受损是PCD和其他疾病预先确定的功能后果 慢性肺部疾病，到目前为止，还没有建立的体外模型能够准确地预测 基因、纤毛运动、MCC和呼吸表型之间的关系，因此许多 基因-表型之间的关系仍不清楚。我们的跨学科研究项目是为 为了解决未得到满足的需求，需要了解a)区域特定的呼吸道组织和b)纤毛病变- 导致基因分型，影响MCC。为了实现这一点，我们将完成特定的目标，旨在：1)使用体外 保留其活体上皮组织的肺组织，作为纤毛呼吸道上皮的模型 综合评价大、小气道生物力学结构-功能关系(目标1)； 使用已建立的人气管-支气管(大)呼吸道体外模型来确定最小的一组 体外纤毛组织和体外纤毛组织之间保守的结构和功能参数(目标2)； 应用最先进的基于物理的计算方法来开发一种能够 预测纤毛组织的结构-功能关系(目标3)和4)使用最小参数集 它将体外和体外模型中的功能性MCC定义为输入到计算机模型中，以预测区域性- 大小气道纤毛缺陷引起的MCC的特殊改变(所有AIMS)。我们的具体目标 建立我们的长期目标，将体外和硅胶模型结合起来，作为临床前、精确的医学 评估小分子或基因编辑疗法的工具，以实现更有针对性和更有效的治疗 以粘液纤毛清除不良为特征的肺部疾病的治疗方案。

PROJECT SUMMARY Mucociliary clearance (MCC) is a critical mechanical defense mechanism of the human respiratory system. Poor MCC is a fundamental feature of many inherited and acquired respiratory diseases including primary ciliary dyskinesia (PCD), asthma, chronic bronchitis, and cystic fibrosis (CF). Due to the complex organization of the lung, it is largely unknown how defects of the ciliary machinery change functional MCC and how regional variability of airway epithelial structure, including cell type proportions and ciliary beat parameters, affect local MCC pathophysiology. These knowledge gaps dramatically impair our ability to predict the degree of pulmonary dysfunction imparted by specific ciliary defects, and to understand the airway region-specific onset observed in many lung diseases. While impaired MCC is a pre-determined functional consequence of PCD and other chronic lung diseases, to date, there is no established in vitro model that is able to accurately predict the relationship between genotype, cilia motility, MCC, and respiratory phenotype and, therefore, many genotype-phenotype relationships remain unexplained. Our transdisciplinary research program is designed to address an unmet need to understand how a) region-specific airway organization and b) ciliopathy- causing genotypes, impact MCC. To achieve this we will complete specific aims designed to: 1) use ex vivo lung tissues, that retain their in vivo epithelial organization, as models of ciliated airway epithelia to comprehensively evaluate biomechanical structure-function relationships in large and small airways (Aim 1); 2) use established in vitro models of the human tracheo-bronchial (large) airways to determine a minimal set of structural and functional parameters that are conserved between in vitro and ex vivo ciliated tissues (Aim 2); 3) apply state-of-the-art physics-based computational approaches to develop an in silico model that will be able to predict structure-function relationships of ciliated tissues (Aim 3) and 4) use the minimal set of parameters that define functional MCC in both in vitro and ex vivo models as input into the in silico model to predict regional- specific changes in MCC due to ciliary defects in both large and small airways (All Aims). Our specific objectives build to our long-term goal of combining in vitro and in silico models as a preclinical, precision medicine tool for evaluating small molecule or gene-editing therapeutics toward more targeted and efficient treatment regimens of pulmonary diseases characterized by poor mucociliary clearance.