Motile cilia project from the airway apical surface and directly interface with inhaled external environment. Due to cilia’s nanoscale dimension and high beating frequency, quantitative assessment of their motility remains a sophisticated task. Here we described a robust approach for reproducible engineering of apical-out airway organoid (AOAO) of defined size. Propelled by exterior-facing cilia beating, the mature AOAO exhibited stable rotational motion when surrounded by Matrigel. We developed a computational framework leveraging computer vision algorithms to quantify AOAO rotation and validated its correlation with direct measurement of cilia motility. We further established the feasibility of using AOAO rotation to recapitulate and measure defective cilia motility caused by chemotherapy-induced toxicity and by CCDC39 mutations in cells from primary ciliary dyskinesia patient. We expect our rotating AOAO model and the associated computational pipeline to offer a generalizable framework to expediate modeling of and therapeutic development for genetic and environmental ciliopathies.

中文翻译：

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工程旋转顶端外气道类器官用于评估呼吸纤毛运动

活动纤毛从气道顶端表面突出并直接与吸入的外部环境接触。由于纤毛的纳米级尺寸和高跳动频率，对其运动性的定量评估仍然是一项复杂的任务。在这里，我们描述了一种稳健的方法，用于定义大小的顶端外气道类器官 (AOAO) 的可重复工程。在向外纤毛跳动的推动下，成熟的 AOAO 在被 Matrigel 包围时表现出稳定的旋转运动。我们开发了一个利用计算机视觉算法来量化 AOAO 旋转的计算框架，并验证了它与直接测量纤毛运动的相关性。我们进一步确定了使用 AOAO 旋转来概括和测量由化疗诱导的毒性和原发性纤毛运动障碍患者细胞中的 CCDC39 突变引起的纤毛运动缺陷的可行性。我们希望我们的旋转 AOAO 模型和相关的计算流程能够提供一个通用的框架，以加快遗传和环境纤毛病的建模和治疗开发。