Modeling emphysematous-tissue related cellular subsets for future use in advanced cell culture models

Abstract

Emphysema entails the irreversible loss of the alveoli. There are no treatments to halt or recover alveolar loss, and drug discovery is hindered by culture systems not replicating the complex disease processes. We aim to develop an Alveolus-on-Chip model with disease-relevant cells to study cellular cross-talk related to impaired tissue repair in emphysema. To this end we developed mediator cocktails to promote emphysema-associated cellular subsets and hallmark processes. We cultured primary alveolar epithelial type-2 cells (AT2) and MRC-5 fibroblasts with emphysema-related compounds (inflammatory mediators TNFα, IL-1β, IFN-γ, and hedgehog (HH) signaling agonists SAG and shh). We assessed AT2 survival, proliferation, and AT1 differentiation, and induction of a reported lung fibroblast phenotype expressing GLI1 and IL7, as well as inflammatory marker IL-8. Exposure of MRC-5 to HH agonists induced GLI1 expression, but inflammatory mediators impair this expression. IL7 gene expression is increased by inflammatory mediators, but this doesn’t translate to higher protein levels; IL-8 protein production is increased by culture with inflammatory mediators. AT2 cultures were negatively affected by culture with inflammatory mediators, showing lower SP-C and AGER expression and viability, but higher IL-8 levels. Culture with emphysema-related mediators results in phenotype changes in both AT2 and fibroblast populations similar to phenotypes detected in human tissue. Further research on these phenotypes will allow us to develop an Alveolus-on-Chip model to study cellular cross-talk underlying emphysema pathology and test regenerative strategies. Supported by the NWO Perspective grant #P12-14.

Authors 6

1. Philip Jalink 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, the Netherlands, Leiden, Netherlands

2. Charlotte J.N. Ovaa 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, the Netherlands, Leiden, Netherlands

3. Eric D.L. Muré 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, the Netherlands, Leiden, Netherlands

4. Pieter Sicco Hiemstra 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, the Netherlands, Leiden, Netherlands

5. Irene Heijink

   University Medical Center Groningen · University of Groningen

   Affiliation as printed

   2University of Groningen, University Medical Center Groningen, Department of Pathology & Medical Biology and Pulmonary Diseases, Groningen Research Institute for Asthma and COPD (GRIAC), Groningen, the Netherlands, Groningen, Netherlands

6. 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, the Netherlands, Leiden, Netherlands