Multicellular modeling of ciliopathy by combining iPS cells and microfluidic airway-on-a-chip technology

Abstract

Mucociliary clearance is an essential lung function that facilitates the removal of inhaled pathogens and foreign matter unidirectionally from the airway tract and is innately achieved by coordinated ciliary beating of multiciliated cells. Should ciliary function become disturbed, mucus can accumulate in the airway causing subsequent obstruction and potentially recurrent pneumonia. However, it has been difficult to recapitulate unidirectional mucociliary flow using human-derived induced pluripotent stem cells (iPSCs) in vitro and the mechanism governing the flow has not yet been elucidated, hampering the proper humanized airway disease modeling. Here, we combine human iPSCs and airway-on-a-chip technology, to demonstrate the effectiveness of fluid shear stress (FSS) for regulating the global axis of multicellular planar cell polarity (PCP), as well as inducing ciliogenesis, thereby contributing to quantifiable unidirectional mucociliary flow. Furthermore, we applied the findings to disease modeling of primary ciliary dyskinesia (PCD), a genetic disease characterized by impaired mucociliary clearance. The application of an airway cell sheet derived from patient-derived iPSCs and their gene-edited counterparts, as well as genetic knockout iPSCs of PCD causative genes, made it possible to recapitulate the abnormal ciliary functions in organized PCP using the airway-on-a-chip. These findings suggest that the disease model of PCD developed here is a potential platform for making diagnoses and identifying therapeutic targets and that airway reconstruction therapy using mechanical stress to regulate PCP might have therapeutic value.

A Biomimetic Human Lung‐on‐a‐Chip with Colorful Display of Microphysiological Breath

“…The lung-on-a-chip has emerged as a promising tool in the past decade for modeling the complexity of human lung structure and function. [1][2][3][4][5][6][7][8] Upon now, a series of lung-on-a-chip devices have been established to recapitulate the lung cells/tissue interactions and study lung diseases such as inflammation, asthma, lung cancer, and lung injuries. [4][5][6][8][9][10][11][12][13][14][15][16][17][18] As these models could resemble human lung in terms of structure, they have significantly advanced our understanding in lung diseases and respiratory drug discovery.…”

Clinically Relevant Influenza Virus Evolution Reconstituted in a Human Lung Airway-on-a-Chip

“…This dynamic mode of drug administration on-chip has been shown to replicate the therapeutic time window of oseltamivir observed in patients in vivo ( 8 ). We and others also have shown that fluid shear stress in the microfluidic human airway chip leads to physiologically relevant mucociliary velocities and coordinated mucociliary movement, rather than random motions, which may play a role in viral entry and clearance ( 5 , 29 ). In addition, infection of these chips with the same strain of influenza H1N1 virus as used in the present study has been shown to elicit a host cytokine response that closely replicates host responses observed in human patients ( 8 ), which also may affect viral replication dynamics.…”

Breathing on Chip: Dynamic flow and stretch tune cellular composition and accelerate mucociliary maturation of airway epitheliumin vitro

“…Although we did not explore the isolated effects of dynamic flow and stretch on airway epithelial biology, the ability to deliver these cues and achieve measurable effects on airway differentiation and homeostasis using a commercially available chip model combined with a robust protocol for primary airway culture opens new avenues for investigating ciliogenesis, airway disease and epithelial regeneration in the presence of mechanical stresses and establish beneficial versus adverse effects. Our method could also be applicable to the differentiation of alveolar or airway cells derived from induced pluripotent stems cells 22,34 as well as multi-tissue airway models and enhance their structural and functional maturation in vitro .…”