Studying lung microvascular endothelial dysfunction upon cigarette smoke exposure using an alveolus-on-chip

Abstract

Lung microvascular endothelial cells are hypothesized to contribute to alveolar repair and regeneration. We recently demonstrated loss of these cells in unaffected tissue regions from patients with emphysema and confirmed their support function for alveolar epithelial cells in vitro. Here, we modelled the cellular cross-talk between these compartments in response to whole cigarette smoke (WCS) using a novel alveolus-on-chip, to identify potential therapeutic targets. Primary endothelial cells, pericytes and alveolar epithelial type-2 cells (AEC2) were isolated from human lung tissue. First, endothelial cells were seeded in a fibrin-collagen hydrogel in an open-top alveolus-on-chip to form vascular networks, alone, or in co-culture with pericytes. AEC2 were next seeded as a monolayer on top of the hydrogel. After confluence was reached, chip cultures were exposed to WCS, after which chips were processed for various analyses. Endothelial cells alone were able to form networks with vessel diameters of 5-10 μm, while in presence of pericytes, networks with a 1-3 μm vessel diameter were observed. Upon whole CS exposure, AEC2 monolayer integrity was maintained while the vascular network disintegrated. Interestingly, when vascular networks were exposed in absence of the AEC2 layer, disintegration was not observed, suggesting that an AEC2-derived component(s) promotes this disintegration. In conclusion, lung microvascular endothelial cells are negatively affected by CS exposure via alveolar epithelial cross-talk. Further studies will focus on identification of this AEC2-derived factor(s) to investigate if it could provide a therapeutic target for prevention of emphysema progression.

Authors 9

1. Abilash Ravi Aachen 1PulmoScience Lab Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   1PulmoScience Lab, Department of Pulmonology, Leiden University Medical Center, Leiden, Netherlands

2. Tarek Gensheimer

   University of Twente

   Affiliation as printed

   2Department of Applied Stem Cell Technologies, University of Twente, Enschede, Netherlands

3. Xinhui Wu

   University of Groningen

   Affiliation as printed

   3Department of Molecular Pharmacology, Faculty of Science and Engineering, University of Groningen, Groningen, Netherlands

4. Jill R. Johnson

   Aston University

   Affiliation as printed

   4School of Biosciences, College of Health & Life Sciences, Aston University, Birmingham, United Kingdom

5. Martin C. Harmsen

   University of Groningen

   Affiliation as printed

   3Department of Molecular Pharmacology, Faculty of Science and Engineering, University of Groningen, Groningen, Netherlands

6. Reinoud Gosens

   University of Groningen

   Affiliation as printed

   3Department of Molecular Pharmacology, Faculty of Science and Engineering, University of Groningen, Groningen, Netherlands

7. Pieter S. Hiemstsra Aachen 1PulmoScience Lab Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   1PulmoScience Lab, Department of Pulmonology, Leiden University Medical Center, Leiden, Netherlands

8. Andries D. van der Meer

   University of Twente

   Affiliation as printed

   2Department of Applied Stem Cell Technologies, University of Twente, Enschede, Netherlands

9. Anne M. van der Does Aachen 1PulmoScience Lab Dept of Pulmonology, Leiden University Medical Center

   Leiden University Medical Center

   Affiliation as printed

   1PulmoScience Lab, Department of Pulmonology, Leiden University Medical Center, Leiden, Netherlands