Engineering rotating apical-out airway organoid for assessing respiratory cilia motility

Piyumi Wijesekara; Prakarsh Yadav; Lydia A Perkins; Donna B Stolz; Jonathan M Franks; Simon C Watkins; Emily Reinoso Jacome; Steven L Brody; Amjad Horani; Jian Xu; Amir Barati Farimani; Xi Ren

doi:10.1016/j.isci.2022.104730

Engineering rotating apical-out airway organoid for assessing respiratory cilia motility

iScience. 2022 Jul 9;25(8):104730. doi: 10.1016/j.isci.2022.104730. eCollection 2022 Aug 19.

Authors

Piyumi Wijesekara¹, Prakarsh Yadav², Lydia A Perkins³, Donna B Stolz⁴, Jonathan M Franks⁴, Simon C Watkins⁴, Emily Reinoso Jacome¹, Steven L Brody⁵, Amjad Horani^{6

7}, Jian Xu⁵, Amir Barati Farimani^{1

2}, Xi Ren^{1

2}

Affiliations

¹ Department of Biomedical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA, USA.
² Department of Mechanical Engineering, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA, USA.
³ Department of Biological Sciences, Carnegie Mellon University, 5000 Forbes Avenue, Pittsburgh, PA, USA.
⁴ Department of Cell Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA.
⁵ Department of Medicine, Washington University School of Medicine, St. Louis, MO, USA.
⁶ Department of Pediatrics, Washington University School of Medicine, St. Louis, MO, USA.
⁷ Department of Cell Biology and Physiology, Washington University School of Medicine, St. Louis, MO, USA.

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.

Grants and funding

R01 HL128370/HL/NHLBI NIH HHS/United States