Periodic Reporting for period 4 - SymBreakOrganoid (Cell-to-cell variability during symmetry breaking in organoid development)
Reporting period: 2023-04-01 to 2023-09-30
The overall objectives of the project are to determine the cause and consequences of cellular heterogeneity in intestinal organoids. We have established novel experimental frameworks for analyzing collective behaviors of cells in tissue and develop perturbation methods to have improved spatio temporal resolution. This is allowing us to understand design principles of tissue development, homeostasis and regeneration. We are also developed tools to perturb individual cells and create synthetic heterogeneities in tissues. This then allowed us to translate some of our finding in human organoids and discover novel mechanisms of human intestinal regeneration.
For the grant we performed imaging multiplexed time course experiments of organoid development for determining the source of cell to cell variability and to predict symmetry breaking events. We then performed scRNAseq and ATACseq time course experiment to understand the extend of cell to cell variability. Finally, we built a light sheet microscope that allows us to follow the formation of organoids from one cell to a fully formed organoid to follow the consequence of cellular heterogeneity. We have implementing the optogenetic perturbations in the light sheet to control cellular heterogeneity.
The work establishes novel paradigm on how to analyzer collective behaviors. We also discovered that organoid development follow a regenerative response and this is having incredible implication in how to study regeneration in vitro, in mouse but also in human. It allowed us to test novel compounds in intestinal regeneration in vivo. Finally, it allowed us to set the standard in the field on how to perform image based analysis in organoids