Team LARGHERO

Stem Cell Biotechnologies

The ZEPHYR project is a translational research program targeting pediatric tracheal replacement using a decellularized tracheal matrix. It relies on an original validated and patented decellularization protocol, comprehensive in vitro characterization, and in vivo studies in pigs focused on bio-integration, revascularization, and re-epithelialization. The ultimate goal is a safe clinical transfer, including cryobanking, GMP manufacturing, and a regulatory framework.

We are also conducting a project on the characterization, culture, and differentiation of tracheal epithelial cells in the context of tracheal re-epithelialization. Our objective is to understand the mechanisms of tissue regeneration, to characterize, isolate, and culture porcine tracheal epithelial cells, and to compare them with human tracheal cells. The long-term goal is to culture and differentiate tracheal epithelial cells under Good Manufacturing Practice (GMP) conditions in order to reconstitute a functional epithelium.

Our objective is to develop cell therapy products composed of human umbilical cord-derived mesenchymal stromal cells (UC-MSCs) to treat immune and/or inflammatory diseases such as graft-versus-host disease, autoimmune diseases, or severe inflammatory syndromes. We are particularly interested in the heterogeneity of the biological properties and immunomodulatory functions of UC-MSCs, which makes it difficult to standardize treatments and predict clinical responses. We are therefore evaluating different strategies to overcome this limitation: (1) characterization of UC-MSCs and their immunomodulatory functions, in order to identify and select optimal donors; (2) development of a cell therapy product composed of a pool of multiple UC-MSC donors including an optimal donor, in order to reduce the variability of the therapeutic effect between individual donors and to increase the immunoregulatory potential of non-optimal donors; and (3) identification of UC-MSC sub-populations exhibiting the highest immunomodulatory functions.

The team’s work focuses on the potential of cell therapy in the antenatal management of myelomeningocele. Animal experiments have demonstrated the benefit of a patch of mesenchymal stromal cells (MSCs) isolated from umbilical cords as an adjunct treatment to in utero surgery. These encouraging results have opened the way to a translational application in humans. Additional ongoing experiments are investigating the use of MSC-derived extracellular vesicles in this indication.

The TREX group concentrates its research efforts on the study of various corneal pathologies — including aniridia, Fuchs endothelial dystrophy, and limbal stem cell deficiency — by combining four complementary approaches: understanding pathophysiological mechanisms, identifying biomarkers, developing or improving innovative therapies, and establishing advanced in vitro models. For its work, the TREX group uses patient samples, primary cells, and cell lines cultured in vitro in 2D or 3D using bioprinting approaches, as well as in vivo models mimicking human pathologies. Finally, the TREX group devotes a significant portion of its work to extracellular vesicles, with a view to therapeutic development.

Our group has been conducting a research program dedicated to the development of tissue-engineered esophageal substitutes for more than fifteen years, in close collaboration with digestive surgery teams and numerous other partners. The experimental approach in porcine models initially evaluated various cellularized substitutes, demonstrating their immunomodulatory and tissue remodeling capacities in vivo. The team subsequently shifted toward the development and use of a decellularized porcine, and then human, esophagus. This acellular substitute demonstrated in vivo the capacity to induce tissue remodeling toward an esophageal phenotype similar to that of cellularized substitutes.

These results led to the obtention of institutional funding and authorization from the ANSM (French National Agency for the Safety of Medicines and Health Products) to initiate the ESOGRAFT clinical trial, supported by the creation of a bank of decellularized and cryopreserved human esophagi. In parallel, we are continuing our translational research with two goals: 1. to adapt this approach for pediatric use, primarily for the treatment of esophageal atresia, and 2. to understand the mechanisms of action underlying the observed tissue regeneration.