Wnt Signaling Separates the Progenitor and Endocrine Compartments during Pancreas Development
Nadav Sharon,
Jordan Vanderhooft,
Juerg Straubhaar,
Jonas Mueller,
Raghav Chawla,
Quan Zhou,
Elise N. Engquist,
Cole Trapnell,
David K. Gifford,
Douglas A. Melton
Affiliations
Nadav Sharon
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA
Jordan Vanderhooft
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA
Juerg Straubhaar
FAS Informatics, Harvard University, Cambridge, MA 02138, USA
Jonas Mueller
Computer Science and Artificial Intelligence Laboratory, MIT, Cambridge, MA 02412, USA
Raghav Chawla
Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Division of Hematology/Oncology, Seattle Children’s Hospital, Seattle, WA 98105, USA; Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA
Quan Zhou
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA
Elise N. Engquist
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA
Cole Trapnell
Department of Genome Sciences, University of Washington, Seattle, WA 98195, USA; Molecular & Cellular Biology Program, University of Washington, Seattle, WA 98195, USA
David K. Gifford
Computer Science and Artificial Intelligence Laboratory, MIT, Cambridge, MA 02412, USA
Douglas A. Melton
Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, Cambridge, MA 02138, USA; Corresponding author
Summary: In vitro differentiation of pluripotent cells into β cells is a promising alternative to cadaveric-islet transplantation as a cure for type 1 diabetes (T1D). During the directed differentiation of human embryonic stem cells (hESCS) by exogenous factors, numerous genes that affect the differentiation process are turned on and off autonomously. Manipulating these reactions could increase the efficiency of differentiation and provide a more complete control over the final composition of cell populations. To uncover in vitro autonomous responses, we performed single-cell RNA sequencing on hESCs as they differentiate in spherical clusters. We observed that endocrine cells and their progenitors exist beside one another in separate compartments that activate distinct genetic pathways. WNT pathway inhibition in the endocrine domain of the differentiating clusters reveals a necessary role for the WNT inhibitor APC during islet formation in vivo. Accordingly, WNT inhibition in vitro causes an increase in the proportion of differentiated endocrine cells. : In vitro differentiation of pluripotent cells into β cells is a promising alternative to cadaveric islet transplantation as a cure for type 1 diabetes. Sharon et al. use scRNA-seq to identify the cell populations that form during the process and uncover a role for WNT pathway inhibition during endocrine differentiation. Keywords: Single-cell RNA sequencing, human embryonic stem cells, hESCs, in vitro differentiation, β cells, insulin, diabetes, pancreas development, WNT pathway, APC
