WP1 will characterize, enhance and produce DP8α regulatory T cells (Tregs) in the GMP format to treat IBD patients. Proteomic approaches have been used to identify the antigen(s) recognized by these DP8α Tregs and derived from the bacteria F. prausnitzii. One GMP-compliant antibody panel was selected to maximize the quality of DP8α Tregs sorting. We are currently performing a deep characterization of DP8α Tregs, finalizing a method to enhance their suppressive activity and setting up conditions to produce them in the GMP format.
WP2 aims to develop a well-characterized library of patient-derived organoids mimicking dysfunctional IBD gut tissue by enriching the epithelial cells with selected immune cells and connecting them to a neural enteric network. Gut samples from non-IBD and IBD patients requiring surgery were collected to establish organoids made only from epithelial intestinal cells (hEIO). Some hEIO individual lines have already been established and stored. Their characterization is ongoing. Purification of immune and neural cells from the gut have started and co-cultures tests with hEIOs are in progress.
WP3 aims to establish the optimal experimental conditions to generate vascularized intestinal “mesoscale” organoids (PDLM) (several organoids fused together) using bioprinting. To develop the ex vivo model of functional and immunocompetent IBD (PDLM), as a proof of concept, we have developed the PDLM from commercial human colonic primary cells. Using the same condition, we establish PDLM derived from healthy and IBD patients. We have begun to develop the vascularization of these organoids, and to integrate autologous T cells.
WP4 aims to develop the first microfluidic device for perfusing a large vascularized system integrating PDLM and lymph node tissue. We successfully developed the first microfluidic chip prototype and hydrogel (PLMA, a photopolymerizable hydrogel obtained from platelets for 3D culture) designed to promote tissue vascularization. The microfluidic device demonstrated biocompatibility, good optical properties, and its ability to effectively confine hydrogel within the central chamber. Meanwhile, the significant progress made in capillary production will be integrated into the on-chip culture model in the next period.
WP5 aims to establish the objective criteria to assess the response of complex gut organoids to gold standard IBD therapy. Monitoring of the organoid growth over time has been performed by videomicroscopy. Characterization of all these organoids is currently being performed by immunofluorescence and qRT-PCR. Identification and functional test of Paneth cells was addressed.
WP6 aims to compare, using complex organoid models of IBD, the impact of DP8α Tregs treatment with the reference therapies evaluated in WP5. It has not really started yet.
WP7 aids project coordination and communication, supporting the MITI² coordinator in managing consortium. It ensures internal communication and disseminates project results to maximize visibility. A dedicated project management team supports the MITI² coordinator in overseeing consortium, ensuring that the project meets its objectives. Moreover, it oversees the dissemination of project results, enhancing visibility and impact.
WP8 objectives are to establish and implement all necessary ethics procedures to ensure that the project fully complies with ethical standards and regulations. It sets out the ethical requirements that the project, financed by the European Commission, must comply with.
