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SUMMARY:Thesis defense - Ihssane Idrissi
DESCRIPTION:Venue : Centre Broca \n\nIhssane Idrissi \nTeam : Quantitative imaging of the cell\nIINS \nTitle\nDevelopment of a toxicology assay using 3D cellular models and soSPIM-based high-content screening \nAbstract\nDrug-induced liver injury (DILI) remains a major cause of drug withdrawal\, highlighting the urgent need for predictive and physiologically relevant in vitro assays. Three-dimensional (3D) cell cultures have emerged as promising models for drug screening and toxicity testing\, as they better reproduce tissue architecture and function compared to conventional two-dimensional (2D) systems. However\, their integration into high-content screening (HCS) still faces important challenges\, including the standardization and parallelization of culture\, the minimization of handling steps between culture and imaging\, and the development of imaging methods capable of capturing complete 3D volumes with high speed and minimal photobleaching and phototoxicity\, while remaining fully compatible with HCS requirements. \nThis PhD project aims to establish a dedicated HCS pipeline for hepatotoxicity assays using HepaRG liver spheroids\, microfabricated JeWells culture devices\, single-objective light-sheet fluorescence microscopy (soSPIM)\, and dedicated analysis tools. The soSPIM architecture\, based on integrated 45° mirrors\, enables high-speed\, low-phototoxic 3D imaging. Combined with JeWells devices\, arrays of truncated pyramidal-shaped cavities\, it allows the parallelized culture and standardized imaging of hundreds of 3D samples within a single chip\, providing quantitative morphological readouts at the single-organoid level. \nAs a proof of concept\, we modeled drug-induced cholestasis on 7-day-old HepaRG spheroids cultured in JeWell devices. Spheroids were exposed to reference compounds\, including chlorpromazine and indomethacin\, imaged on the soSPIM-HCS platform after immunolabelling\, and analyzed for nuclear\, cytoskeletal\, and hepatobiliary transporter markers. We demonstrated the ability of this platform to capture both morphological disruptions and functional impairments characteristic of cholestatic injury in liver organoids. \nOverall\, this work establishes a methodological framework integrating standardized 3D culture\, advanced 3D imaging\, and automated analysis into a single HCS pipeline for hepatotoxicity assessment. Beyond the case study of cholestasis\, this platform paves the way for broader applications of 3D organoid-based assays in drug discovery and toxicology. \nKeywords: Light-sheet microscopy\, HepaRG spheroids\, High-Content Screening\, Hepatotoxicity\, Cholestasis\, Quantitative image analysis. \nJury\nJean-Baptiste Sibarita  – Ingénieur de Recherche\, Université de Bordeaux\, CNRS – Directeur de Thèse \nCharlotte Rivière – Directrice de Recherche\, Université de Lyon\, CNRS – Rapporteure \nCorinne Lorenzo –  Ingénieure de Recherche\, Université de Toulouse\, CNRS – Rapporteure \nPierre Nassoy – Directeur de Recherche\, Université de Bordeaux\, CNRS – Examinateur \nPascale Roux – Lab Manager\, Sanofi – Invitée \nRémi Galland – Chargé de Recherche\, Université de Bordeaux\, CNRS – Invité \n
URL:https://www.bordeaux-neurocampus.fr/en/event/thesis-defense-ihssane-idrissi/
CATEGORIES:Thesis
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