Uncovering the molecular and cellular heterogeneity of pancreatic ductal cells

Data suggest that pancreatic ductal cells have a latent capacity for regeneration but the molecular regulation of this potential has been completely unexplored. I have recently optimised ex vivo pancreatic organoids as an efficient and tractable system to study the effect of perturbations on ductal cell fate.

I propose to use organoids to characterise the cell types coexisting in the ductal compartment that are permissive or non-permissive for changing cell fate and the molecular pathways involved. Differentiation will be induced by Ngn3/Pdx1/MafA overexpression or by a medium containing differentiation-inducing factors. I will use transcriptomics to delineate different cell types in ductal INS-GFP organoids to identify markers of reprogramming-competent cells. I will also compare RNASeq profiles of reprogrammed, intermediate and unchanged organoid cells to reveal genes required for cell fate change and perform a Crispr/Cas9-based loss of function screen to identify the crucial signaling pathways involved in the maintenance of duct cell fate, using CK19-cherry as a marker of ductal cell fate.

These results will elucidate the fundamental biology of pancreatic ductal cell plasticity and provide clues for how to unlock the regenerative capacity of pancreatic ductal cells in vivo for future therapies.