WNT-mediating TCF/LEF transcription factor gene expression in early human pluripotency and cell lineages differs from the rodent paradigm

Connor Ross; Paula A. Balestrini; Lawrence E. Bates; Takuya Azami; Taiye Asakole; Maxine Semple; Marika Salonna; Richard Gyuris; Jennifer Nichols; Norah Fogarty; Stefan Hoppler

doi:10.1242/jcs.264257

WNT-mediating TCF/LEF transcription factor gene expression in early human pluripotency and cell lineages differs from the rodent paradigm

Connor Ross
, Paula A. Balestrini
, Lawrence E. Bates
, Takuya Azami
, Taiye Asakole
, Maxine Semple
, Marika Salonna
, Richard Gyuris
, Jennifer Nichols
, Norah Fogarty
, Stefan Hoppler^* (Corresponding Author)

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

Embryonic stem (ES) cell research has uncovered different requirements for WNT/β-catenin signalling in human naïve pluripotent cells compared to the mouse paradigm. It is therefore important to study WNT/β-catenin signalling directly in models that recapitulate early human development. Since TCF/LEF transcription factors mediate regulation of target genes downstream of WNT/β-catenin signalling, we examined the regulation, expression and protein localisation of the four TCF/LEF genes by analysing in vitro ‘snapshots’ of human development, leveraging naïve and primed pluripotent cells, blastoids and preimplantation blastocysts. Strikingly, we comprehensively confirm clear differences between mouse and human pluripotent stem cells, suggesting their differential requirements for WNT signalling reflects a pluripotent state-dependent manner. Human naïve ES cells express considerably lower levels of TCF7L1, unlike their mouse counterparts. TCF7L2 is robustly expressed in the trophectoderm derived from naïve ES cells, in blastoids and human preimplantation blastocysts. In primed pluripotent stem cells, active WNT/β-catenin signalling induces the expression of both TCF7 and LEF1, concomitant with hallmark gastrulation markers. The expression of human TCF/LEF genes indicates a differential requirement for WNT/β-catenin signalling throughout early human embryo development that warrants further investigation.

Original language	English
Article number	jcs264257
Number of pages	19
Journal	Journal of Cell Science
Volume	138
Issue number	18
Early online date	29 Sept 2025
DOIs	https://doi.org/10.1242/jcs.264257
Publication status	Published - Sept 2025

Bibliographical note

Acknowledgements
We would like to thank all members of the Hoppler, Fogarty and Nichols groups for critical feedback and helpful discussions on this project and Yvonne Turnbull for lab management. We thank Dr Debbie Wilkinson (Aberdeen Advanced Microscopy and Histology), Ann Wheeler (Edinburgh Imaging) and Darran Clements (Cambridge Imaging) for their assistance with confocal microscopy. We thank Dr Andrea Holme and Robert Pineda (Ian Fraser Cytometry Centre, University of Aberdeen) for their kind assistance with flow cytometry. We thank all the clinical staff and patients who have kindly donated their embryos for research purposes and the clinical staff who coordinate the transfer of donated embryos to research projects.

Data Availability Statement

All relevant data and details of resources can be found within the article and its supplementary information.

Funding

S.H. and C.R. are supported by a Biotechnology and Biological Sciences Research Council (BBSRC) research grant (BB/Y001974/1). C.R. was supported by a Medical Research Council doctoral training partnership (MRC-DTP) PhD studentship (2117379). J.N. is supported by a BBSRC research grant (BB/Y005120/1). N.E.F. is supported by a UK Research and Innovation (UKRI) Future Leaders Fellowship (MR/W006693/1). Open Access funding provided by University of Aberdeen. Deposited in PMC for immediate release.

Funders	Funder number
Biotechnology and Biological Sciences Research Council	BB/Y001974/1, BB/Y005120/1
Medical Research Council	2117379
UK Research and Innovation	MR/W006693/1

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

SDG 3 Good Health and Well-being

Keywords

Human development
Naïve pluripotency
WNT/β-catenin signalling
TCF/LEF transcription factors
Extra-embryonic cell lineages
Early embryonic cell lineages

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Final published version, 20.3 MBLicence: CC BY

Iain Fraser Cytometry Centre
Duncan, L. (Senior Application Scientist), Laird, A. (Technician) & Burgoyne, K. (Technician)
Institute of Medical Sciences
Research Facilities: Facility
Microscopy and Histology
Wilkinson, D. (Manager) & Milne, G. (Manager)
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Cite this

Ross, C., Balestrini, P. A., Bates, L. E., Azami, T., Asakole, T., Semple, M., Salonna, M., Gyuris, R., Nichols, J., Fogarty, N., & Hoppler, S. (2025). WNT-mediating TCF/LEF transcription factor gene expression in early human pluripotency and cell lineages differs from the rodent paradigm. Journal of Cell Science, 138(18), Article jcs264257. https://doi.org/10.1242/jcs.264257

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title = "WNT-mediating TCF/LEF transcription factor gene expression in early human pluripotency and cell lineages differs from the rodent paradigm",

abstract = "Embryonic stem (ES) cell research has uncovered different requirements for WNT/β-catenin signalling in human na{\"i}ve pluripotent cells compared to the mouse paradigm. It is therefore important to study WNT/β-catenin signalling directly in models that recapitulate early human development. Since TCF/LEF transcription factors mediate regulation of target genes downstream of WNT/β-catenin signalling, we examined the regulation, expression and protein localisation of the four TCF/LEF genes by analysing in vitro {\textquoteleft}snapshots{\textquoteright} of human development, leveraging na{\"i}ve and primed pluripotent cells, blastoids and preimplantation blastocysts. Strikingly, we comprehensively confirm clear differences between mouse and human pluripotent stem cells, suggesting their differential requirements for WNT signalling reflects a pluripotent state-dependent manner. Human na{\"i}ve ES cells express considerably lower levels of TCF7L1, unlike their mouse counterparts. TCF7L2 is robustly expressed in the trophectoderm derived from na{\"i}ve ES cells, in blastoids and human preimplantation blastocysts. In primed pluripotent stem cells, active WNT/β-catenin signalling induces the expression of both TCF7 and LEF1, concomitant with hallmark gastrulation markers. The expression of human TCF/LEF genes indicates a differential requirement for WNT/β-catenin signalling throughout early human embryo development that warrants further investigation.",

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author = "Connor Ross and Balestrini, \{Paula A.\} and Bates, \{Lawrence E.\} and Takuya Azami and Taiye Asakole and Maxine Semple and Marika Salonna and Richard Gyuris and Jennifer Nichols and Norah Fogarty and Stefan Hoppler",

note = "Acknowledgements We would like to thank all members of the Hoppler, Fogarty and Nichols groups for critical feedback and helpful discussions on this project and Yvonne Turnbull for lab management. We thank Dr Debbie Wilkinson (Aberdeen Advanced Microscopy and Histology), Ann Wheeler (Edinburgh Imaging) and Darran Clements (Cambridge Imaging) for their assistance with confocal microscopy. We thank Dr Andrea Holme and Robert Pineda (Ian Fraser Cytometry Centre, University of Aberdeen) for their kind assistance with flow cytometry. We thank all the clinical staff and patients who have kindly donated their embryos for research purposes and the clinical staff who coordinate the transfer of donated embryos to research projects.",

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AU - Ross, Connor

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AU - Azami, Takuya

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AU - Semple, Maxine

AU - Salonna, Marika

AU - Gyuris, Richard

AU - Nichols, Jennifer

AU - Fogarty, Norah

AU - Hoppler, Stefan

N1 - Acknowledgements We would like to thank all members of the Hoppler, Fogarty and Nichols groups for critical feedback and helpful discussions on this project and Yvonne Turnbull for lab management. We thank Dr Debbie Wilkinson (Aberdeen Advanced Microscopy and Histology), Ann Wheeler (Edinburgh Imaging) and Darran Clements (Cambridge Imaging) for their assistance with confocal microscopy. We thank Dr Andrea Holme and Robert Pineda (Ian Fraser Cytometry Centre, University of Aberdeen) for their kind assistance with flow cytometry. We thank all the clinical staff and patients who have kindly donated their embryos for research purposes and the clinical staff who coordinate the transfer of donated embryos to research projects.

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N2 - Embryonic stem (ES) cell research has uncovered different requirements for WNT/β-catenin signalling in human naïve pluripotent cells compared to the mouse paradigm. It is therefore important to study WNT/β-catenin signalling directly in models that recapitulate early human development. Since TCF/LEF transcription factors mediate regulation of target genes downstream of WNT/β-catenin signalling, we examined the regulation, expression and protein localisation of the four TCF/LEF genes by analysing in vitro ‘snapshots’ of human development, leveraging naïve and primed pluripotent cells, blastoids and preimplantation blastocysts. Strikingly, we comprehensively confirm clear differences between mouse and human pluripotent stem cells, suggesting their differential requirements for WNT signalling reflects a pluripotent state-dependent manner. Human naïve ES cells express considerably lower levels of TCF7L1, unlike their mouse counterparts. TCF7L2 is robustly expressed in the trophectoderm derived from naïve ES cells, in blastoids and human preimplantation blastocysts. In primed pluripotent stem cells, active WNT/β-catenin signalling induces the expression of both TCF7 and LEF1, concomitant with hallmark gastrulation markers. The expression of human TCF/LEF genes indicates a differential requirement for WNT/β-catenin signalling throughout early human embryo development that warrants further investigation.

AB - Embryonic stem (ES) cell research has uncovered different requirements for WNT/β-catenin signalling in human naïve pluripotent cells compared to the mouse paradigm. It is therefore important to study WNT/β-catenin signalling directly in models that recapitulate early human development. Since TCF/LEF transcription factors mediate regulation of target genes downstream of WNT/β-catenin signalling, we examined the regulation, expression and protein localisation of the four TCF/LEF genes by analysing in vitro ‘snapshots’ of human development, leveraging naïve and primed pluripotent cells, blastoids and preimplantation blastocysts. Strikingly, we comprehensively confirm clear differences between mouse and human pluripotent stem cells, suggesting their differential requirements for WNT signalling reflects a pluripotent state-dependent manner. Human naïve ES cells express considerably lower levels of TCF7L1, unlike their mouse counterparts. TCF7L2 is robustly expressed in the trophectoderm derived from naïve ES cells, in blastoids and human preimplantation blastocysts. In primed pluripotent stem cells, active WNT/β-catenin signalling induces the expression of both TCF7 and LEF1, concomitant with hallmark gastrulation markers. The expression of human TCF/LEF genes indicates a differential requirement for WNT/β-catenin signalling throughout early human embryo development that warrants further investigation.

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SN - 0021-9533

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JO - Journal of Cell Science

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M1 - jcs264257

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WNT-mediating TCF/LEF transcription factor gene expression in early human pluripotency and cell lineages differs from the rodent paradigm

Abstract

Bibliographical note

Data Availability Statement

Funding

UN SDGs

Keywords

Access to Document

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Equipment

Iain Fraser Cytometry Centre

Microscopy and Histology

Cite this