Background: IL11 was recently reported to induce the formation of myofibroblasts and their secretion of collagen, and its subcutaneous administration drives the development of pulmonary fibrosis in mice. While the role of IL11 on myofibroblast differentiation is described, less is known as to how IL11 affects epithelial-mesenchymal communication associated with epithelial repair. Furthermore, it is not yet well-established which cell types are the main producers and responders to IL11 in vivo.

Here we aim to investigate the role of IL11 in alveolar regeneration.

Methods: Epithelial progenitor cell behavior was studied using an organoid model, comprised of a co-culture of primary mouse CD31-CD45-Epcam+ cells with either CCL206 fibroblasts or primary human fibroblasts. After 14 days of growth, the number and size of the organoids were determined. Additionally, immunofluorescent staining for Surfactant protein C (SPC) and acetylated tubulin (ACT) was performed to define alveolar vs airway organoids. Immunohistochemical staining of IL11 and IL11 receptor was performed on human lung tissue to observe producer and responder cells of IL11 signaling.

Results: The presence of rhIL11 significantly decreased the number of both mouse and human organoids formed in a concentration-dependent manner. Additionally, the fraction of SPC expressing organoids was concentration-dependently reduced by rhIL11 whereas the fraction of SPC-ACT- organoids was increased. Organoid size was not affected by rhIL11. Finally, immunohistochemistry revealed that IL11R is found in the smooth muscle, airway epithelium, endothelium and macrophages, whereas IL11 was primarily observed in the airway epithelium, alveolar type 2 cells and macrophages.

Conclusions: IL11 may impair epithelial progenitor function by suppressing their activation and by suppressing the formation of mature alveolar cell types. IL11R localization suggests mesenchymal activation by IL11 may contribute to the observed effects on progenitor function, possibly through mesenchyme-epithelial crosstalk.

IL11 negatively impacts adult lung alveolar organoid formation

Rationale: In chronic obstructive pulmonary disease (COPD), the airways are characterized by goblet cell hyperplasia, decreased ciliated cell numbers and an altered turnover of the extracellular matrix (ECM). Current 2D (co-)culture systems lack the spatial in vivo environment that is formed by ECM. In this study we aimed to develop a lung ECM hydrogel-based co-culture model to investigate bidirectional interactions between fibroblasts and epithelial cells in relation to ECM changes. 

Methods: Porcine lungs were decellularized, lyophilized and ground to a powder. This powder was porcine pepsin-solubilized for 48h, pH neutralized, buffered with PBS and incubated at 37°C for 30 min to generate ECM hydrogels. Human primary lung fibroblasts (50,000/ml) from non-COPD donors were mixed with hydrogels prior to gelation in transwells. The next day, the hydrogels were coated with collagen type IV or left uncoated. After 24h, 100,000 primary bronchial epithelial cells (PBEC) from non-COPD donors were seeded on top of fibroblast seeded and unseeded gels. The cells were cultured submerged for 4 days and subsequently at air liquid interface for 14 days. The hydrogels were imaged using inverted light microscopy and by histology on paraffin-embedded sections

Results: PBECs attached and formed a mono layer on the collagen-IV coated and uncoated hydrogels. All gels contracted after 48 hours. The absence of fibroblasts appeared to reduce the degree of contraction. After 4 days of submerged culture the epithelial layer formed a stacked multicellular confluent layer on the hydrogels. After 14 days of air exposure the multicellular epithelial cells on the hydrogels further expanded and the fibroblasts remodeled their environment, as shown by a change in structure and staining intensity around the fibroblasts in the ECM hydrogel seen in the H&E staining.

Conclusions: The combination of a lung ECM hydrogel with a basement membrane like coating made it possible to co-culture primary lung fibroblasts and PBECs in 3D at ALI, generating a morphology resembling the in vivo airway wall. 


Airway epithelial cells, fibroblasts and the extracellular matrix: a ménage à trois resembling the airway wall in vivo


INTRODUCTION: Idiopathic Pulmonary Fibrosis (IPF) is characterized by aberrant ECM deposition and remodeling of the lung – structural changes which further drive a pro-fibrotic cellular microenvironment. Decellularized lung ECM-derived hydrogels resemble the mechanical properties of native decellularized tissues, potentially providing a 3D model mimicking native cell-ECM interactions. We aimed to characterize this 3D human lung microenvironment model, with respect to stiffness and viscoelastic properties, in the presence and absence of primary human lung fibroblasts.
METHODS: Lyophilized powders of decellularized IPF and control lung matrices (pooled from 6 donors per group) were pepsin digested, and formed to hydrogels seeded with control primary lung fibroblasts (n=6 donors), and cultured for 14 days. Stiffness and viscoelastic relaxation were measured by Low-Load Compression Testing (20% strain). 
RESULTS: IPF hydrogels showed a trend towards higher stiffness compared to control hydrogels and cell-seeded IPF hydrogels were significantly stiffer than cell-seeded controls (1.85 ± 0.48 kPA vs 1.04 ± 0.22 kPA, p=0.0183, Kruskal-Wallis test) (Fig. 1). Time to reach 100% viscoelastic relaxation was similar in cell-seeded compared to native hydrogels for both IPF and control hydrogels. Total percentage relaxation of cell-seeded IPF hydrogels was significantly lower than of cell-seeded control hydrogels (108.4% ± 2.39% vs 117.6% ± 3.63%, p=0.0495, Kruskal-Wallis test) while empty hydrogels reached comparable values. The percentage relaxation was in general lower in IPF hydrogels than control hydrogels (cell-seeded or not). 
DISCUSSION & CONCLUSIONS: The mechanical properties of the ECM hydrogels were modified by fibroblasts and the source of the ECM influenced these modifications. These data suggest that higher stiffnesses and altered relaxation patterns of the ECM may be involved in fibrotic responses in IPF. Fibroblast-seeded ECM-derived hydrogels can provide more insight on cell-ECM interactions 


Fibroblast-seeded lung extracellular matrix- derived hydrogels as an in vitro model for stromal bed in  Idiopathic Pulmonary Fibrosis (IPF)

poster

[![](https://assets.underline.io/uploads/markdown_image/1/image/261902fdf187a88bfa74da226ff77186.png)](https://drive.google.com/file/d/1gUViQJJHCSgM38OhwpDoTdN89evCsZ-7/view?usp=sharing)
[![](https://assets.underline.io/uploads/markdown_image/1/image/1605292c232e6f185c7f87ee38e8502a.png)](https://drive.google.com/file/d/1kPkznVX_6cx-n846z_pUtyVDZql9KImc/view?usp=sharing)


### **Welcome Message**

Welcome to the inaugural **Dutch Lung Congress.**  We have all missed our opportunities to network and interact with our colleagues in the last twelve months or more.  While our desire was to meet face to face once again, when it became evident that this would not be possible until much later in 2021 (or even into 2022) the hard working committee organizing the Dutch Lung Congress found, what we hope will be, an excellent alternative to facilitate our annual meeting.

The Dutch Lung Congress aims to bring together everyone interested in research related to pulmonary diseases in the Netherlands.  We hope this event will provide opportunities for multidisciplinary interactions to enhance our approaches for addressing these challenging diseases.  Each session will include input from patients, clinical and basic and/or translational scientists discussing the latest information around the central themes chosen by the organizing committee.  One advantage of holding our congress as a virtual event this year is that we will have the opportunity to hear from world leading experts from all over the globe.  We hope you enjoy the programme the organizing committee have put together and will engage in dynamic discussions with all the speakers.

One highlight of our congress will be the Young Investigator Session.  Remember to join us for this exciting session when we will hear from the presenters who submitted the top scoring abstracts.  This promises to be a session full of exciting science that will display the strengths that constitute the future talent in our field.

When we are not listening to these great presentations we can all meet in Gathertown.  Within this interactive platform you will be able to meet colleagues and friends for networking. Enjoy a private chat over a virtual coffee, visit the booths of our sponsors and other parties, chat with the Young Investigator Board members or take some timeout in the relaxation spaces.  While in Gathertown take the opportunity to visit all the diverse posters on display. Chat live with the presenters or leave them a question on the discussion board for them to engage with you.  Make sure you check for any messages each time you enter Gathertown to make the most of your networking opportunities.

We look forward to seeing you during the DLC and the great scientific (and other) discussions we are anticipating during our 2021 meeting.

On behalf of the DLC organizing Committee
<br>**Janette Burgess
<br>Chair, DLC Organizing Committee**
![](https://assets.underline.io/uploads/markdown_image/1/image/f461fb275d9667fb2a76a30813388e90.png)



### Special Session: Publieksmiddag
![](https://assets.underline.io/uploads/markdown_image/1/image/77e768dd500e6205175e5cafba59abdc.png)






![](https://assets.underline.io/uploads/markdown_image/1/image/f461fb275d9667fb2a76a30813388e90.png)
**More about the committee team:**:
[Barbro N. Melgert, PhD] (https://drive.google.com/file/d/1q_IIH3JvTivumCUzzsZGX7BgD41HxQmk/view?usp=sharing)
// [Bas Holverda, PhD] (https://drive.google.com/file/d/1QKXHmTTQjgs4OZVuM8doXU055S-fu_uU/view?usp=sharing)
// [Frits Franssen, PhD] (https://drive.google.com/file/d/1d7HRUJaqGuRFzkPN3SVgkbBfqFkDkWq2/view?usp=sharing)
// [Janette Burgess, PhD] (https://drive.google.com/file/d/1VHFlrEN6uCQGeeW3Lj4I8aTw7HsB0GFr/view?usp=sharing)
// [Michel van den Heuvel, PhD] (https://drive.google.com/file/d/1JeOEP8MHxiK_4lONr8UM2HaxdxpkAmA2/view?usp=sharing)
// [Patrick Bhairosing] (https://drive.google.com/file/d/1xI8ORq-g1bbVcrlRDviZ9NhAwanc4dFl/view?usp=sharing)
// [Gimano Amatngalim, PhD] (https://drive.google.com/file/d/1fyVDDKARLNjRk_JZk8_GyhGMjmp6DWHp/view?usp=sharing)
// [Peter Kunst, PhD] (https://drive.google.com/file/d/1QhtB4nQezpAjbfUAcBnINc-Bm3Shy2Lf/view?usp=sharing)
// [Dr. Michel van den Heuvel, Prof., MD, PhD] (https://drive.google.com/file/d/1JBdNEBXZCEikEZhJ6MoD5DhQfXMz3bP6/view?usp=sharing)
![](https://assets.underline.io/uploads/markdown_image/1/image/f461fb275d9667fb2a76a30813388e90.png)

Please register with DLC to be able to enter the conference on Underline.io. If you have already registered, please login to Underline.io to be able to see full conference content. In case you do not have an Underline account, please create one. Thank you 

Janette Burgess

3

Presentations

IL11 negatively impacts adult lung alveolar organoid formation

Airway epithelial cells, fibroblasts and the extracellular matrix: a ménage à trois resembling the airway wall in vivo

Fibroblast-seeded lung extracellular matrix- derived hydrogels as an in vitro model for stromal bed in  Idiopathic Pulmonary Fibrosis (IPF)

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Janette Burgess

3

Presentations

IL11 negatively impacts adult lung alveolar organoid formation

Airway epithelial cells, fibroblasts and the extracellular matrix: a ménage à trois resembling the airway wall in vivo

Fibroblast-seeded lung extracellular matrix- derived hydrogels as an in vitro model for stromal bed in Idiopathic Pulmonary Fibrosis (IPF)

Stay up to date with the latest Underline news!

PRESENTATIONS

CONFERENCES

COMPANY

RESOURCES

Fibroblast-seeded lung extracellular matrix- derived hydrogels as an in vitro model for stromal bed in  Idiopathic Pulmonary Fibrosis (IPF)