Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Stephanie Brown
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Epigenetics Programme, The Babraham Institute, Cambridge, United Kingdom
Shota Nakanoh
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom; Division of Embryology, National Institute for Basic Biology, Okazaki, Japan
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Brandon T Wesley
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Daniele Muraro
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom; Wellcome Sanger Institute, Hinxton, Cambridge, United Kingdom
A Sophie Brumm
Human Embryo and Stem Cell Laboratory, The Francis Crick Institute, London, United Kingdom
Human Embryo and Stem Cell Laboratory, The Francis Crick Institute, London, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge, United Kingdom
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Daniel Ortmann
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Department of Surgery, University of Cambridge, Cambridge, United Kingdom
Wellcome–MRC Cambridge Stem Cell Institute, Jeffrey Cheah Biomedical Centre, University of Cambridge, Cambridge, United Kingdom; Epigenetics Programme, The Babraham Institute, Cambridge, United Kingdom; Centre for Trophoblast Research, University of Cambridge, Cambridge, United Kingdom
The signalling pathways that maintain primed human pluripotent stem cells (hPSCs) have been well characterised, revealing a critical role for TGFβ/Activin/Nodal signalling. In contrast, the signalling requirements of naïve human pluripotency have not been fully established. Here, we demonstrate that TGFβ signalling is required to maintain naïve hPSCs. The downstream effector proteins – SMAD2/3 – bind common sites in naïve and primed hPSCs, including shared pluripotency genes. In naïve hPSCs, SMAD2/3 additionally bind to active regulatory regions near to naïve pluripotency genes. Inhibiting TGFβ signalling in naïve hPSCs causes the downregulation of SMAD2/3-target genes and pluripotency exit. Single-cell analyses reveal that naïve and primed hPSCs follow different transcriptional trajectories after inhibition of TGFβ signalling. Primed hPSCs differentiate into neuroectoderm cells, whereas naïve hPSCs transition into trophectoderm. These results establish that there is a continuum for TGFβ pathway function in human pluripotency spanning a developmental window from naïve to primed states.
