Dataset Information

Expression data from cells sorted from human fetal pancreas

ABSTRACT: Although a large set of data is available concerning organogenesis in animal models, information remains scarce on human organogenesis. In this work, we performed temporal mapping of human fetal pancreatic organogenesis using cell surface markers. We demonstrate that in the human fetal pancreas at 7 weeks of development, the glycoprotein 2 (GP2) marks a multipotent cell population that will differentiate either into the acinar, ductal and endocrine lineages. Development towards the acinar lineage is paralleled by a substantial increase in GP2 expression. Conversely, a subset of the multipotent GP2+ population undergoes endocrine differentiation by down-regulating GP2 and CD142 and turning on NEUROG3, an early marker of endocrine differentiation. Endocrine maturation will progress by up-regulating SUSD2 and lowering ECAD levels. Finally, we show that in vitro differentiation of pancreatic endocrine cells derived from human pluripotent stem cells mimics key in vivo events. Our work constitutes a powerful approach to more precisely define intermediate cell population during conversion of multipotent progenitors into the 3 main human pancreatic cell types (acinar, ductal and endocrine) in vivo. As such, the data pave the way to extend our understanding of the origin of mature human pancreatic cell types and how such lineage decisions are regulated.

ORGANISM(S): Homo sapiens

PROVIDER: GSE96697 | GEO | 2017/03/17

SECONDARY ACCESSION(S): PRJNA379384

REPOSITORIES: GEO

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Project description:Although a large set of data is available concerning organogenesis in animal models, information remains scarce on human organogenesis. In this work, we performed temporal mapping of human fetal pancreatic organogenesis using cell surface markers. We demonstrate that in the human fetal pancreas at 7 weeks of development, the glycoprotein 2 (GP2) marks a multipotent cell population that will differentiate either into the acinar, ductal and endocrine lineages. Development towards the acinar lineage is paralleled by a substantial increase in GP2 expression. Conversely, a subset of the multipotent GP2+ population undergoes endocrine differentiation by down-regulating GP2 and CD142 and turning on NEUROG3, an early marker of endocrine differentiation. Endocrine maturation will progress by up-regulating SUSD2 and lowering ECAD levels. Finally, we show that in vitro differentiation of pancreatic endocrine cells derived from human pluripotent stem cells mimics key in vivo events. Our work constitutes a powerful approach to more precisely define intermediate cell population during conversion of multipotent progenitors into the 3 main human pancreatic cell types (acinar, ductal and endocrine) in vivo. As such, the data pave the way to extend our understanding of the origin of mature human pancreatic cell types and how such lineage decisions are regulated. While the mechanisms by which glucose regulates insulin secretion from pancreatic beta cells are now well described, the way glucose modulates gene expression in such cells needs more understanding. Here, we demonstrate that MondoA, but not its paralogue ChREBP, is the predominant glucose-responsive transcription factor in human pancreatic beta EndoC-βH1 cells and in human islets. In high glucose conditions, MondoA shuttles to the nucleus where it is required for the induction of the glucose-responsive genes ARRDC4 and TXNIP, the latter being a protein strongly linked to beta-cell dysfunction and diabetes. Importantly, increasing cAMP signaling in human beta cells, using forskolin or the GLP-1 mimetic Exendin-4, inhibits the shuttling of MondoA and potently inhibits TXNIP and ARRDC4 expression. Furthermore, we demonstrate that modulating MondoA expression functionally affects glucose uptake. These results highlight MondoA as a novel target in beta cells that coordinates transcriptional response to elevated glucose levels.

Project description:Our team has previously characterized a subpopulation of ductal progenitor cells with progenitor-like characteristics. These cells proliferate in response to stimulation of the BMP pathway by BMP-7 and, following withdrawal of this growth factor, differentiate into all three adult epithelial pancreatic lineages (ductal, acinar and endocrine), both in vitro and after transplantation. scRNAseq analysis of sorted human pancreatic ductal cells has confirmed the existence of progenitor-like clusters whose transcriptomic signature suggests a role for stress-mediated de-differentiation in the acquisition of their stemness. Importantly, the application of cell trajectory analyses upon integration of this ductal dataset with other published datasets that also include endocrine and acinar cells revealed the potential existence of ducto-acinar and ducto-endocrine differentiation axes. However, these conclusions were qualified by the fact that they came from the amalgamation of multiple datasets from many different donors, processed and analyzed in different ways, and also in the absence of a defined regenerative stimulus whose effect could be investigated in a truly continuous manner. We hypothesized that the sequential scRNAseq of same-donor HPSs would address the above constraints and allow us, for the first time, to dissect cell fate changes at the single cell level in response to BMP-7 in a “whole human pancreas” setting. To test this hypothesis, we conducted the longitudinal scRNAseq analysis of human pancreatic slices (HPSs) subjected to a 10-day regimen consisting of 5 days of exposure to BMP-7 followed by 5 days without. The most immediately apparent observation is the transcriptomic heterogeneity of the ductal and acinar compartments, which most earlier analyses (based on isolated islet samples that at best contained a small proportion of ductal and acinar cell types) have failed to detect. With the exception of a recent single nucleus RNAseq report, no other similar study thus far has presented the field with a true whole-pancreas analysis where the different cell types of the organ retain their natural proportion. Integration of single-cell datasets obtained from same-donor HPSs at different time points, or in the presence or absence of treatment, allowed us to conduct cell trajectory analyses using true temporal information, rather than by pseudotemporal inferences. This integration revealed population shifts consistent with BMP-7-mediated progenitor cell activation, the blurring of ductal/acinar boundaries, the formation of clear ducto-acinar-endocrine differentiation axes and, notably, the appearance of transitional insulin-producing cell populations. These findings contrast with the pattern observed in untreated controls, whose cluster composition and distribution remain largely unchanged with time –another observation that further confirms the robustness of the HPS model to faithfully mirror the cellular makeup of the human pancreas over a 10-day period. In summary, our study provides the first longitudinal scRNAseq analysis of whole human pancreatic tissue, mapping true cell fate trajectories in a manner that was simply not possible prior to the development of long-term HPS culture techniques. These experiments unequivocally confirm pancreatic plasticity in a dynamic fashion and validate the reliability of this novel human-based model for countless other research applications.

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Expression data from cells sorted from human fetal pancreas

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