MADMpancreas · Using the precision mouse genetic tool MADM to elucidate the role of EGFR in directing beta cell differentiation and pancreatic morphogenesis
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2019-06-01 → 2021-05-31
- EU contribution
- €207,312
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
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Results in brief
Using the precision mouse genetic tool MADM to elucidate the role of EGFR in directing beta cell differentiation and pancreatic morphogenesis
The pancreas houses two physiologically distinct compartments within one organ. The hormone-producing endocrine cells of the islets and the tubular ductal system of the exocrine pancreas share a common developmental progenitor. Recent work from the Semb group implicated Egfr signaling as a critical commitment event that both transcriptionally primes cells for endocrine differentiation–by upregulating Neurog3 expression in metastable progenitors—while also facilitating islet morphogenesis—by antagonizing cellular polarity and inducing delamination from the bipotent trunk epithelium. A central objective of this project was to understand how Egfr signaling is regulated in the developing pancreas. Specifically, we wanted to test the hypothesis that quantitative differences in pathway activation explain why metastable Neurog3-expressing cells commit to endocrinogenesis in a seemingly stochastic and temporally dispersed manner. We further test if such quantitative differences facilitate selection of the different hormone producing fates. To quantitatively manipulate Egfr levels at single-cell resolution, this project employs the mouse genetic tool Mosaic Analysis with Double Markers (MADM).
Data: CORDIS, © European Union
Project objective
Insulin producing beta cells are derived from bipotent pancreatic progenitors (bi-PPs) that constitute the primitive ductal epithelium. While we now know a great deal about the transcriptional networks required for endocrinogenesis and commitment to the beta lineage, considerably less is understood about the cellular events that initiate delamination and activate differentiation networks. The host group recently found that ligand-specific Egfr signaling is essential for these processes. Signaling by the highly potent Egfr ligand betacellulin diminishes apicobasal polarity leading to upregulation of Ngn3, delamination, and beta cell differentiation. The overall aim of this proposal is to elucidate the role of Egfr signaling in the beta cell differentiation process. I will ask: Is the seemingly stochastic differentiation of bi-PPs regulated by inherent differential Egfr signaling? Do quantitative differences in this single pathway lead to different cellular outcomes? How does the targeted loss of Egfr in Ngn3-expressing endocrine precursors affect their differentiation? What are the genes regulated by Egfr pathway activation that putatively commit Ngn3+ progenitors to a beta cell fate? And how does the disruption of endocrine differentiation affect ductal morphogenesis? Answers to these questions have remained elusive because of the difficulty of observing individual cell behaviors in situ in the developing pancreas. To meet this challenge, I will adapt the cutting edge mouse genetic tool Mosaic Analysis with Double Markers (MADM) to quantitatively manipulate Egfr levels in bi-PPs and to concomitantly label altered cells. The successful implementation of MADMouse will deliver findings of immediate interest to investigators seeking to improve the directed differentiation of therapeutic grade beta cells from hPSCs. I will augment my expertise in precision mouse genetics with training in hESC culture enhancing my career goal of establishing my own research group.
Original text from CORDIS.
Participants
- KOBENHAVNS UNIVERSITET · KOBENHAVNCoordinatorDenmark
Links
Data: CORDIS, © European Union