Grapin-Botton Group: Endoderm and Pancreas Development
Our aim is to understand how progenitor cells of the pancreas read architectural cues to choose between expansion and differentiation. We use these principles to recapitulate the process in vitro from stem cells.
More specifically, Professor Grapin-Botton and her group investigate the impact of the cellular and organ architecture on the cells’ fate choices and how single cells act in a community to generate an organ. To do so, they use mouse genetics, live imaging in 3D and they developed 3D in vitro “organoid” culture systems modelling development. More recently they used human in vitro stem cell models investigate human development. These studies are intended to gain insight into human syndromes impairing pancreas development and they guide the generation of replacement beta cells for Diabetes therapy.
3D culture models of pancreas development: we have developed systems to expand pancreas progenitors from a few cells or single cells in 3D in vitro (protocol). This leads to the formation of organoids similar to a pancreas in vitro. We use these in vitro models together with in silico modelling to understand how different cell types with a specific spatial arrangement are created from one cell or an initially homogeneous group of cells. We are adapting these systems to control the differentiation of beta cells from stem cells to build a model of human pancreas development and disease. Using genetic modifications of human stem cells we can test how perturbations of genes associated with Diabetes risk cause the disease.
Contributions of individual cells to organ community: During pancreas development progenitors proliferate, expanding the size of the organ while a subset of them differentiates to generate the endocrine cells that regulate glucose levels and acinar cells contributing to food digestion. Using clonal analysis, live imaging and single cell PCR and sequencing, we study how each cell makes such decisions, the role of stochastic processes and logic rules. By understanding the process we hope to control it and generate numerous beta cells for diabetes therapy.
Architectural cues controlling cell differentiation and proliferation: The differentiation of pancreatic cells occurs largely from polarized progenitors aligned in ducts. Using in vivo investigations in mice we are investigating whether this architectural environment and more specifically planar cell polarity controls the differentiation of pancreatic cell types. To do so, we are investigating in which cell types this pathway is active, how it is organized in space and the role of genes controlling the architecture in syndromes associating kidney cysts and diabetes. We recently found that pancreatic cells secrete pancreatic juice from very early during development which may create a flow controlling the organization of the ductal tree and possibly differentiation.
Dahl-Jensen, S.B., Yennek, S., Flasse, L., Larsen, H.L., Sever, G.K., Novak, I., Sneppen K. and Grapin-Botton, A. (2018). Deconstructing the principles of ductal network formation in the pancreas. PLoS Biology, 16, e2002842, doi:10.1371/journal.pbio.2002842.
Ramond, C., Beydag-Tasöz, B.S., Azad, A., van de Bunt, M., Petersen, M.K.B., Beer, , N.L., Glaser, N., Berthault, C., Gloyn, A.L., Hansson, M., McCarthy, M.I., Honoré, C., Grapin-Botton, A. and Scharfmann, R. (2018). Understanding human fetal pancreas development using subpopulation sorting, RNA sequencing and single-cell profiling. Development 145, dev.165480, doi:10.1242/dev.165480.
Larsen, H.L., Martín-Coll, L., Nielsen, A.V., Wright, C.V.E., Trusina, A., Kim, Y.H. # and Grapin-Botton, A. # (2017). Stochastic priming and spatial cues orchestrate heterogeneous clonal contribution to mouse pancreas organogenesis. Nature Communications 8, 605, doi:10.1038/s41467-017-00258-4. # Shared last authorship
Petersen, M.B.K., Azad, A., Ingvorsen, C., Hess, K., Hansson, M., Grapin-Botton, A.# and Honoré, C.# (2017). Single-Cell Gene Expression Analysis of a Human ESC Model of Pancreatic Endocrine Development Reveals Different Paths to β-Cell Differentiation. Stem Cell Reports 9, 1246-1261. doi:10.1016/j.stemcr.2017.08.009. * Equal contribution. # Shared last authorship
Kim, Y. H., List Larsen, H., Rué, P., Lemaire, L.A., Ferrer, J. and Grapin-Botton, A. (2015). Cell cycle-dependent differentiation dynamics balances growth and endocrine differentiation in the pancreas. PLOS Biology 13:e1002111, doi:10.1371/journal.pbio.1002111.
Greggio, C., De Franceschi, F., Figueiredo-Larsen, M., Gobaa, S., Ranga, A., Semb, H., Lutolf, M. and Grapin-Botton, A. (2013). Artificial three-dimensional niches deconstruct pancreas development in vitro. Development 140, 4452-4462, doi: 10.1242/dev.096628.
Cortijo, C., Gouzi, M., Tissir, F. and Grapin-Botton, A. (2012). Planar cell polarity controls pancreatic beta cell differentiation and glucose homeostasis. Cell Reports 2, 1593-1606, doi:10.1016/j.celrep.2012.10.016.
Pancreas progenitors in 3D
Dispersed cells from the embryonic pancreas are grown in Matrigel TM and expand within a week into a branched organ. The organoid can be visualized by the cell nuclei expressing GFP (Green Fluorescent Protein) under the control of the Pdx1 promoter, an essential transcription factor dynamically expressed during pancreatic development.