Aurore DUQUENOY
Convention CIFRE entre la société MaaT Pharma et l’IRT Bioaster
Directeur de thèse : Dr Vincent THOMAS
Co-Directeur de thèse : Dr Joël DORE
Vincent.THOMAS@bioaster.org
Microbiota Program
Campus de l’Institut Pasteur, Bâtiment François Jacob – 3e étage
28, rue du Docteur Roux
75015 – PARIS, FRANCE
http://www.bioaster.org/en/
Expansion contrôlée du microbiote fécal en vue d’une utilisation en transplantation fécale autologue
MaaT Pharma est une entreprise qui développe des solutions standardisées de microbiothérapie (transplantation fécale) dans le cadre de dysbioses entéropathiques. La société a développé un procédé pour conditionner les suspensions fécales par congélation. Ces suspensions de selles ont été testées in vivo chez la souris pour évaluer leur capacité à reconstruire le microbiote original. MaaT Pharma a obtenu les autorisations pour réaliser des premiers essais cliniques chez l’homme par administration rectale. Elle s’est par la suite intéressée à l’étude de la multiplication du microbiote intestinal via un procédé de fermentation adapté capable de reproduire les microbiotes fécaux et par conséquent générer des quantités supplémentaires de matériels à des fins thérapeutiques.
C’est dans ce contexte que le projet de thèse s’inscrit ; réalisé sous forme de convention Cifre avec l’Institut de Recherche Technologique Bioaster, le projet consiste à investiguer les conditions de culture qui vont permettre de multiplier de manière contrôlée le microbiote fécal en vue d’une utilisation thérapeutique. Les premiers objectifs ont été de mettre en place un système de fermentation in vitro permettant d’amplifier et de maintenir le microbiote fécal ainsi que de développer des outils analytiques (notamment par cytométrie en flux) pour le suivi de l’évolution des écosystèmes au cours du processus de fermentation.
L’élaboration du procédé de fermentation s’est axée sur le choix des systèmes ainsi que l’évaluation des conditions de culture afin d’optimiser l’expansion du microbiote fécal. Ces premiers essais ont permis de dégager et fixer des paramètres importants du procédé permettant d’atteindre les objectifs fixés à savoir le maintien de la richesse et de la diversité ainsi que le maintien de communautés d’intérêts.
La mise au point de ces conditions a nécessité l’étude de nombreux échantillons en parallèle. La cytométrie en flux, permettant l’analyse rapide et multiparamétrique d’échantillons, a été implémentée pour suivre l’évolution des écosystèmes complexes au cours du processus de fermentation. Les marquages en cytométrie en flux mis au point permettent d’évaluer la proportion de bactéries Gram+/Gram- dans des échantillons complexes. Cet outil est utilisé pour caractériser les produits issus du processus de fermentation et pourrait être utilisé en routine pour qualifier des produits issus de fermentations industrielles.
Des travaux complémentaires sont en cours sur le développement d’outils analytiques additionnels par cytométrie en flux afin d’approfondir la caractérisation des échantillons complexes. Les effets immuno-modulateurs de souches d’intérêts du microbiote intestinal sont actuellement investigués sur des lignées cellulaires immunitaires et épithéliales.
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