Anterior-Posterior Patterning of Definitive Endoderm Generated from Human Embryonic Stem Cells Depends on the Differential Signaling of Retinoic Acid, Wnt-, and BMP-Signaling.
As known from model organisms, such as frog, fish, mouse, and chicken, the anterior-posterior patterning of the definitive endoderm (DE) into distinct domains is controlled by a variety of signaling interactions between the DE and its surrounding mesoderm. This includes Wnt/FGFs and BMPs in the posterior half and all-trans-retinoic acid, TGF-β-ligands, Wnt-, and BMP-inhibitors in the anterior half of the DE sheet. However, it is currently unclear how these embryonic tissue interactions can be translated into a defined differentiation protocol for human embryonic stem cells. Activin A has been proposed to direct DE into a SOX2-positive foregut-like cell type. Due to the pleiotropic nature of SOX2 in pluripotency and developing cells of the foregut, we purified DE-cells by magnetic cell sorting and tested the effects of anteriorizing and posteriorizing factors on pure endoderm. We show in contrast to previous studies that the generation of the foregut marked by SOX2/FOXA2 double-positive cells does not depend on activin A/TGF-β-signaling but is mediated by the inhibition of Wnt- and BMP-signaling. Retinoic acid can posteriorize and at the same time dorsalize the foregut toward a PDX1-positive pancreatic duodenal cell type whereas active Wnt/beta-catenin signaling synergistically with FGF-2, BMP-4, and RA induces the formation of CDX2-positive posterior endoderm. Thus, these results provide new insights into the mechanisms behind cell specification of human DE derived from pluripotent stem cells. Stem Cells 2016;34:2635-2647.
Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
Claudia
C
Davenport
Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
Ulf
U
Diekmann
Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
Insa
I
Budde
Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
Nora
N
Detering
Institute of Clinical Biochemistry, Hannover Medical School, Hannover, Germany.
Ortwin
O
Naujok
D028341:Activins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D019521:Body Patterning / Q000235:genetics; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D002454:Cell Differentiation; D002460:Cell Line; D004707:Endoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental; D051542:Hepatocyte Nuclear Factor 3-beta / Q000235:genetics / Q000378:metabolism; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D018189:Immunomagnetic Separation; D008648:Mesoderm / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D055748:SOXB1 Transcription Factors / Q000235:genetics / Q000378:metabolism; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D014212:Tretinoin / Q000494:pharmacology; D060449:Wnt Signaling Pathway; D060508:Wnt3 Protein / Q000235:genetics* / Q000378:metabolism / Q000494:pharmacology; D051176:beta Catenin / Q000235:genetics / Q000378:metabolism
C525809:BMP4 protein, human; D055415:Bone Morphogenetic Protein 4; C495270:CTNNB1 protein, human; C496502:FOXA2 protein, human; D018398:Homeodomain Proteins; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D015534:Trans-Activators; D016212:Transforming Growth Factor beta; C555507:WNT3 protein, human; D060508:Wnt3 Protein; C422405:activin A; D051176:beta Catenin; C084010:pancreatic and duodenal homeobox 1 protein; D016222:Fibroblast Growth Factor 2; D028341:Activins; D051542:Hepatocyte Nuclear Factor 3-beta; D014212:Tretinoin
Anterior-posterior patterning; Definitive endoderm; Differentiation; Human embryonic stem cells; Pancreatic duodenal endoderm