Inflammatory Vesseloid ON chip – IVEON

Inflammation is the resulting cellular and tissue response to an aggression, whether due to an infection (bacterial or viral), an injury, an auto-immune disease, or a yet unidentified agent (Behçet’s disease). Blood vessels are in first line when an inflammatory process is triggered because they are the main conveyor of proinflammatory molecules and cues, and are the location of extravasation of leukocytes, fluids and plasma proteins. They also propagate and amplify the immune response by secreting pro-inflammatory molecules that will act on the surrounding tissue. The endothelium which constitutes the inner lining of the vessels is the key player in these processes. Endothelial cell (EC) biology has been extremely well studied in vitro and in vivo. However, 2D in vitro models do not capture the spatial organization of vessels and the complexity of an in vivo environment. Thus, the challenge resides in the development of an inflamed 3D vessel model. This model should be ideally suited for exploring vessel inflammation and endothelitis to better identify the complex cellular and molecular events implicated in these processes. Furthermore, it may represent an ideal platform for testing the effect of pharmacological modulators and drugs.
This project aims at setting up a semiologic model of the inflamed blood vasculature. This is based on major advances in vascular tissue engineering made by the partners of this project, the development of the vesseloid. For this project, the vesseloid will first be stabilized to constitute the building block for an inflamed vasculature and then the EC-dependent inflammatory response will be investigated (cytokine-dependent EC activation, EC and leukocyte interaction after cytokine-activation, effect of EC injury by mechanical trauma, laser ablation, or UV damage) as well as the effect of pharmacological modulation. The project includes the use of state-of-the-art imaging and molecular techniques which will provide detailed mechanistical insights. This project musters a multidisciplinary team, expert in vascular biology, microfluidics, live imaging and molecular approaches. Thus, the IVEON project will deliver a physiopathomimetic organoid for the evaluation of a range of endothelial-dependent inflammatory responses which can be further used for the validation of pharmacological and therapeutic molecules.

In sum, the declared objectives are, thus, as follows:

1- Buiding an Inflammatory vesseloid-on-chip
We will first recreate an enriched environment and complexify the cellular composition. We will also implement this objective by producing vascular cells from IPSC. Furthermore, flow will be introduced as another parameter.

2- Characterizing the 3D model at a molecular and functional level
We will first ascertain that the vesseloid-on-chip exhibit all the features of an inflamed vessel by gene expression profile secretome analysis, cellular bioichemistry, immunostaining and imaging using super-resolution microscopy. We will visualize paracrine-induced autocrine factors secretion with a newly available strategy for protein labelling in living cells and study transmigration of leukocytes in our system.

3- Addressing the biological relevance of the model
We will first address the effect of vascular injury by physical manipulation: and investigate the pharmacological/therapeutic modulation of the vesseloid-on-chip.