Quantitative analysis of mucociliary clearance in airway ciliopathies

项目摘要 粘膜纤毛清除（MCC）是人体呼吸系统的重要机械防御机制。 低MCC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，纤毛机制的缺陷如何改变功能性MCC以及区域性MCC如何改变功能性MCC在很大程度上是未知的。 气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结核程度的能力。 功能障碍所赋予的特定纤毛缺陷，并了解气道区域特异性发病观察， 许多肺部疾病。虽然受损的MCC是PCD和其他疾病的预先确定的功能后果， 慢性肺部疾病，迄今为止，还没有建立能够准确预测 基因型，纤毛运动，MCC和呼吸表型之间的关系，因此，许多 基因型-表型关系仍然无法解释。我们的跨学科研究计划旨在 为了解决未满足的需要，以了解a）区域特异性气道组织和B）纤毛病变- 导致基因型影响MCC。为了实现这一目标，我们将完成设计的具体目标：1）使用体外 肺组织，其保留其体内上皮组织，作为纤毛气道上皮的模型， 综合评价大小气道的生物力学结构-功能关系（目的1）; 2） 使用已建立的人体气管-支气管（大）气道体外模型， 在体外和离体纤毛组织之间保守的结构和功能参数（目的2）; 3） 应用最先进的基于物理学的计算方法来开发一个计算机模型， 预测纤毛组织的结构-功能关系（目的3）和4）使用最小参数集 将体外和离体模型中的功能性MCC定义为计算机模拟模型的输入，以预测区域- 由于大气道和小气道中的纤毛缺陷导致的MCC的特定变化（所有目的）。我们的具体目标 建立我们的长期目标，将体外和计算机模型结合起来，作为临床前精准医学 用于评估小分子或基因编辑疗法以实现更有针对性和更有效的工具 以粘膜纤毛清除差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.