GSE134179
TrxG complex catalytic and non-catalytic activity play distinct roles in pancreas progenitor specification and differentiation (E13.5 Dpy30?P pancreas scRNA-Seq)
Purpose: To investigate whether pancreas progenitor specification may be altered, we performed single-cell RNA-sequencing (scRNA-seq) using E13.5 control and Dpy30?P pancreas. To determine whether Dpy30?P cells have a different developmental trajectory than control cells, we performed pseudo-temporal analysis (Trapnell et al., 2014). Methods: Floxed Dpy30 mice were crossed to Pdx1-Cre driver mice to obtain conditional deletion of Dpy30 exon 4 in the pancreas. In all studies, knockout mice (Dpy30?P, Pdx1-Cre; Dpy30flox/flox) were compared to littermate controls (Dpy30flox/flox or Dpy30flox/wt). Results: In total, we sequenced 3,900 control and 3,044 Dpy30?P cells from which 786 control and 477 Dpy30?P epithelial cells were identified for further analysis. As expected, control progenitor cells gave rise to cells in two distinct branches corresponding to the acinar and endocrine lineages, with terminal markers upregulated in cells at the ends of these branches. Dpy30?P progenitors differentiated along these same trajectories; however, relatively few Dpy30?P cells were found at the ends of these branches, and acinar and endocrine terminal marker expression was reduced in Dpy30?P cells. Interestingly, a higher fraction of Dpy30?P cells were found within the MPC/BPC and acinar clusters, predominantly at the expense of endocrine cells. Analyzing the fraction of cells in G1 phase across pseudo-time showed that a higher fraction of Dpy30?P acinar lineage cells were in G1 phase. Conclusions: Overall, this suggests that MPC/BPCs and acinar cells may have a prolonged G1 phase due to a failure to appropriately induce genes essential for progression through G2M or S phase. Overall design: Single-cell RNA-sequencing was performed using one E13.5 control and one E13.5 Dpy30?P pancreas