Small airway-on-a-chip enables analysis of human lung inflammation and drug responses in vitro.

Kambez H Benam,Thomas C Ferrante,Geraldine A Hamilton,Cedric Hubeau,Anthony Bahinski,Antonio Varone,Michael Salmon,Stephen E Alves,Donald E Ingber,Carolina Lucchesi,James C Weaver,Hyun-Hee Lee,Remi Villenave

doi:10.1038/nmeth.3697

Abstract

Here we describe the development of a human lung 'small airway-on-a-chip' containing a differentiated, mucociliary bronchiolar epithelium and an underlying microvascular endothelium that experiences fluid flow, which allows for analysis of organ-level lung pathophysiology in vitro. Exposure of the epithelium to interleukin-13 (IL-13) reconstituted the goblet cell hyperplasia, cytokine hypersecretion and decreased ciliary function of asthmatics. Small airway chips lined with epithelial cells from individuals with chronic obstructive pulmonary disease recapitulated features of the disease such as selective cytokine hypersecretion, increased neutrophil recruitment and clinical exacerbation by exposure to viral and bacterial infections. With this robust in vitro method for modeling human lung inflammatory disorders, it is possible to detect synergistic effects of lung endothelium and epithelium on cytokine secretion, identify new biomarkers of disease exacerbation and measure responses to anti-inflammatory compounds that inhibit cytokine-induced recruitment of circulating neutrophils under flow.

Highlights

The epithelium formed from cells isolated from the small airway epithelium of lungs from both normal humans (Fig. 1g,i and Supplementary Movie 2) and chronic obstructive pulmonary disease (COPD) patients (Fig. 1h) contained many ciliated epithelial cells as well as mucus-producing Goblet cells (Fig. 1g,h), Club cells (Supplementary Fig. 1b) and Basal cells that appeared in strikingly similar proportions to those found in normal human lung (Table 1)
Electron microscopic analysis of the normal lung small airway-on-a-chip confirmed that the cilia that appeared on the apical surface of the polarized epithelium (Fig. 1i) exhibited the same structure (9+2 microtubule organization) and length (~ 6 μm) (Supplementary Fig. 1c) as healthy cilia found in living human lung in vivo[21, 22]
When mucociliary transport was measured by introducing fluorescent polystyrene microbeads into the top channel, rapid coordinated movement of the beads were observed along the surface of the epithelium (Fig. 1k and Supplementary Movie 4) as a result of active synchronized cilia beating; again, the particle velocity measured (50–100 μm/sec) was nearly identical to that observed in the normal human lung airway (Table 1)

Summary

Introduction

Advances in microsystems engineering have recently made it possible to create biomimetic microfluidic cell culture devices, known as ‘organs-on-chips’, that contain continuously perfused microchannels lined by living human cells that recapitulate the multicellular architectures, tissue-tissue interfaces, physicochemical microenvironments and vascular perfusion of the body[17], which potentially offer new opportunities for disease modeling and drug efficacy assessment[18] We previously used this approach to successfully reconstitute the alveolar-capillary interface of the human lung air sac and associated inflammatory responses in vitro[19, 20]. We demonstrate that this in vitro tool provides a way to dissect contributions of individual tissue types (epithelium, endothelium and immune cells) to these organ-level responses, and to identify new anti-inflammatory therapies

Methods

Results

Conclusion