@article{9b6a81f66e084b9c8bbe865f0e6ecfb4,
title = "Engineering rotating apical-out airway organoid for assessing respiratory cilia motility",
abstract = "Motile cilia project from the airway apical surface and directly interface with inhaled external environment. Owing 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) from a defined number of cells. 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 correlated it with the 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 patients with primary ciliary dyskinesia. We expect our rotating AOAO model and the associated computational pipeline to offer a generalizable framework to expedite the modeling of and therapeutic development for genetic and environmental ciliopathies.",
keywords = "Bioengineering, Cell biology, Molecular physiology",
author = "Piyumi Wijesekara and Prakarsh Yadav and Perkins, {Lydia A.} and Stolz, {Donna B.} and Franks, {Jonathan M.} and Watkins, {Simon C.} and {Reinoso Jacome}, Emily and Brody, {Steven L.} and Amjad Horani and Jian Xu and {Barati Farimani}, Amir and Xi Ren",
note = "Funding Information: This work was supported by the Department of Defense Peer-Reviewed Medical Research Program W81XWH2110183 (X.R. and A.B.F.), and the DSF Charitable Foundation (X.R.). We are grateful to Misti West for laboratory management, to Ming Sun and Mara L. Sullivan from the Center for Biologic Imaging at the University of Pittsburgh for assistance with TEM sample processing, and to Kian Patel for proofreading the article. † These authors contributed equally. P.W. P.Y. A.B.F. and X.R. contributed to the overall design of the project, data interpretation, and preparation of the article. P.W. and P.Y. performed the experiments and analyzed the data. L.A.P. and S.C.W. performed image acquisition regarding cilia beating. D.B.S and J.M.F. performed electron microscopy studies. E.R.J. assisted with figure preparation. S.L.B. A.H. and J.X. provided patient-derived human airway basal stem cells and advised on experiments related to these cells. A patent has been filed by Carnegie Mellon University on the technology described in this study. Funding Information: This work was supported by the Department of Defense Peer-Reviewed Medical Research Program W81XWH2110183 (X.R. and A.B.F.), and the DSF Charitable Foundation (X.R.). We are grateful to Misti West for laboratory management, to Ming Sun and Mara L. Sullivan from the Center for Biologic Imaging at the University of Pittsburgh for assistance with TEM sample processing, and to Kian Patel for proofreading the article. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = aug,
day = "19",
doi = "10.1016/j.isci.2022.104730",
language = "English",
volume = "25",
journal = "iScience",
issn = "2589-0042",
number = "8",
}