Publications by year
In Press
Stirparo GG, Boroviak T, Guo G, Nichols J, Smith A, Bertone P (In Press). Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human preimplantation epiblast.
Abstract:
Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human preimplantation epiblast
AbstractSingle-cell profiling techniques create opportunities to delineate cell fate progression in mammalian development. Recent studies provide transcriptome data from human preimplantation embryos, in total comprising nearly 2000 individual cells. Interpretation of these data is confounded by biological factors such as variable embryo staging and cell-type ambiguity, as well as technical challenges in the collective analysis of datasets produced with different sample preparation and sequencing protocols. Here we address these issues to assemble a complete gene expression time course spanning human preimplantation embryogenesis. We identify key transcriptional features over developmental time and elucidate lineage-specific regulatory networks. We resolve post hoc cell-type assignment in the blastocyst, and define robust transcriptional prototypes that capture epiblast and primitive endoderm lineages. Examination of human pluripotent stem cell transcriptomes in this framework identifies culture conditions that sustain a naïve state pertaining to the inner cell mass. Our approach thus clarifies understanding both of lineage segregation in the early human embryo and of in vitro stem cell identity, and provides an analytical resource for comparative molecular embryology.
Abstract.
Dattani A, Huang T, Smith A, Guo G (In Press). Suppression of YAP Safeguards Human Naïve Pluripotency.
Abstract:
Suppression of YAP Safeguards Human Naïve Pluripotency
ABSTRACTPropagation of human naïve pluripotent stem cells (nPSCs) requires inhibition of MEK/ERK signalling. However, MEK/ERK inhibition also induces differentiation into trophectoderm (TE). Therefore, robust self-renewal requires active suppression of TE fate. Tankyrase inhibition using XAV939 has been shown to stabilise human nPSCs. Here we dissect the mechanism of this effect. Tankyrase inhibition blocks canonical Wnt/β-catenin signalling. However, nPSCs depleted of β-catenin remain dependent on XAV939. We show that XAV939 prevents TE induction by suppressing YAP activity independent of β-catenin. Tankyrase inhibition stabilises angiomotin, which reduces nuclear translocation of YAP1/TAZ. Upon deletion of Angiomotin-family members AMOT and AMOTL2, nuclear YAP increases and XAV939 fails to prevent TE induction. Conversely, nPSCs lacking YAP1 fail to undergo TE differentiation and sustain efficient self-renewal without XAV939. These findings explain the distinct requirement for tankyrase inhibition in human but not mouse naïve PSCs and highlight the pivotal role of YAP in human naïve pluripotency.
Abstract.
2023
Richards D, Cockerell A, Wright L, Dattani A, Guo G, Smith A, Tsaneva K (2023). Biophysical models of early mammalian embryogenesis. Stem Cell Reports, 18, 26-46.
2022
Strawbridge SE, Clarke J, Guo G, Nichols J (2022). Deriving Human Naïve Embryonic Stem Cell Lines from Donated Supernumerary Embryos Using Physical Distancing and Signal Inhibition.
Methods Mol Biol,
2416, 1-12.
Abstract:
Deriving Human Naïve Embryonic Stem Cell Lines from Donated Supernumerary Embryos Using Physical Distancing and Signal Inhibition.
Until recently, naïve pluripotent stem cell lines were not captured from human embryos because protocols were based upon those devised for murine embryonic stem cells. In contrast with early lineage segregation in mouse embryos, human hypoblast specification is not solely dependent upon FGF signaling; consequently, its maturation during embryo explant culture may provide inductive signals to drive differentiation of the epiblast. To overcome this potential risk, here we describe how cells of the immature inner cell mass of human embryos can be physically separated during derivation, achieved via "immunosurgery", to eliminate the trophectoderm, followed by disaggregation of the remaining inner cell mass cells. A modification of a culture regime developed for propagation of human pluripotent stem cells reset to the naïve state is used, which comprises serum-free medium supplemented with various inhibitors of signaling pathways, polarization, and differentiation. Colonies arising from the first plating of an inner cell mass may be pooled for ease of handling, or propagated separately to allow establishment of clonal human naïve embryonic stem cell lines.
Abstract.
Author URL.
Dattani A, Huang T, Liddle C, Smith A, Guo G (2022). Suppression of YAP safeguards human naïve pluripotency.
Development,
149(24).
Abstract:
Suppression of YAP safeguards human naïve pluripotency.
Propagation of human naïve pluripotent stem cells (nPSCs) relies on the inhibition of MEK/ERK signalling. However, MEK/ERK inhibition also promotes differentiation into trophectoderm (TE). Therefore, robust self-renewal requires suppression of TE fate. Tankyrase inhibition using XAV939 has been shown to stabilise human nPSCs and is implicated in TE suppression. Here, we dissect the mechanism of this effect. Tankyrase inhibition is known to block canonical Wnt/β-catenin signalling. However, we show that nPSCs depleted of β-catenin remain dependent on XAV939. Rather than inhibiting Wnt, we found that XAV939 prevents TE induction by reducing activation of YAP, a co-factor of TE-inducing TEAD transcription factors. Tankyrase inhibition stabilises angiomotin, which limits nuclear accumulation of YAP. Upon deletion of angiomotin-family members AMOT and AMOTL2, nuclear YAP increases and XAV939 fails to prevent TE induction. Expression of constitutively active YAP similarly precipitates TE differentiation. Conversely, nPSCs lacking YAP1 or its paralog TAZ (WWTR1) resist TE differentiation and self-renewal efficiently without XAV939. These findings explain the distinct requirement for tankyrase inhibition in human but not in mouse nPSCs and highlight the pivotal role of YAP activity in human naïve pluripotency and TE differentiation. This article has an associated 'The people behind the papers' interview.
Abstract.
Author URL.
Dattani A, Guo G (2022). The people behind the papers – Anish Dattani and Ge Guo.
Development (Cambridge),
149(24).
Abstract:
The people behind the papers – Anish Dattani and Ge Guo
The ability to propagate human naïve pluripotent stem cells (nPSCs) has allowed researchers to address questions about early development that were not previously accessible. The culture conditions themselves have provided insights into the signals required for self-renewal; now, a paper published in Development describes the pathway downstream of XAV939, a tankyrase inhibitor, required for maintenance of human nPSCs but not mouse embryonic stem cells (ESCs). We caught up with first author Anish Dattani and corresponding author Ge Guo, a group leader at the Living Systems Institute, University of Exeter, to find out more about their research.
Abstract.
2021
Stirparo GG, Smith A, Guo G (2021). Cancer-Related Mutations Are Not Enriched in Naive Human Pluripotent Stem Cells.
Cell Stem Cell,
28(1), 164-169.e2.
Abstract:
Cancer-Related Mutations Are Not Enriched in Naive Human Pluripotent Stem Cells.
Previous analysis of RNA sequencing (RNA-seq) data from human naive pluripotent stem cells reported multiple point "mutations" in cancer-related genes and implicated selective culture conditions. We observed, however, that those mutations were only present in co-cultures with mouse feeder cells. Inspection of reads containing the polymorphisms revealed complete identity to the mouse reference genome. After we filtered reads to remove sequences of mouse origin, the actual incidence of oncogenic polymorphisms arising in naive pluripotent stem cells is close to zero.
Abstract.
Author URL.
Guo G, Stirparo GG, Strawbridge SE, Spindlow D, Yang J, Clarke J, Dattani A, Yanagida A, Li MA, Myers S, et al (2021). Human naive epiblast cells possess unrestricted lineage potential.
Cell Stem Cell,
28(6), 1040-1056.e6.
Abstract:
Human naive epiblast cells possess unrestricted lineage potential.
Classic embryological experiments have established that the early mouse embryo develops via sequential lineage bifurcations. The first segregated lineage is the trophectoderm, essential for blastocyst formation. Mouse naive epiblast and derivative embryonic stem cells are restricted accordingly from producing trophectoderm. Here we show, in contrast, that human naive embryonic stem cells readily make blastocyst trophectoderm and descendant trophoblast cell types. Trophectoderm was induced rapidly and efficiently by inhibition of ERK/mitogen-activated protein kinase (MAPK) and Nodal signaling. Transcriptome comparison with the human embryo substantiated direct formation of trophectoderm with subsequent differentiation into syncytiotrophoblast, cytotrophoblast, and downstream trophoblast stem cells. During pluripotency progression lineage potential switches from trophectoderm to amnion. Live-cell tracking revealed that epiblast cells in the human blastocyst are also able to produce trophectoderm. Thus, the paradigm of developmental specification coupled to lineage restriction does not apply to humans. Instead, epiblast plasticity and the potential for blastocyst regeneration are retained until implantation.
Abstract.
Author URL.
Yanagida A, Spindlow D, Nichols J, Dattani A, Smith A, Guo G (2021). Naive stem cell blastocyst model captures human embryo lineage segregation.
Cell Stem Cell,
28(6), 1016-1022.e4.
Abstract:
Naive stem cell blastocyst model captures human embryo lineage segregation.
Human naive pluripotent cells can differentiate into extraembryonic trophectoderm and hypoblast. Here we describe a human embryo model (blastoid) generated by self-organization. Brief induction of trophectoderm leads to formation of blastocyst-like structures within 3 days. Blastoids are composed of three tissue layers displaying exclusive lineage markers, mimicking the natural blastocyst. Single-cell transcriptome analyses confirm segregation of trophectoderm, hypoblast, and epiblast with high fidelity to the human embryo. This versatile and scalable system provides a robust experimental model for human embryo research.
Abstract.
Author URL.
2020
Guo G, Stirparo GG, Strawbridge S, Yang J, Clarke J, Li MA, Myers S, Özel BN, Nichols J, Smith A, et al (2020). Trophectoderm Potency is Retained Exclusively in Human Naïve Cells.
Abstract:
Trophectoderm Potency is Retained Exclusively in Human Naïve Cells
SUMMARY Classical mouse embryology has established a sequence of lineage bifurcations underpinning early mammalian development. Consistent with this paradigm, mouse embryonic stem cells have lost the capacity to generate extraembryonic trophectoderm. We show here, however, that human naïve epiblast stem cells readily produce this lineage. Inhibition of ERK signalling, fundamental to naïve cell propagation, is unexpectedly instrumental in trophectoderm induction. Transcriptome analyses authenticate trophoblast fate and expose a trajectory via reversion to inner cell mass (ICM). Nodal inhibition enhances differentiation and BMP signalling is not engaged. Strikingly, after formative transition primed stem cells cannot make trophectoderm but respond to BMP and form amnion. Gene perturbations in naïve cells reveal that YAP and TFAP2C promote trophectoderm as in mouse, while NANOG suppresses distinctively in human. Finally, ICMs from expanded human blastocysts efficiently regenerate trophectoderm. Thus in human, retained trophectoderm potential is an integral feature of emergent pluripotency that confers higher regulative plasticity.
Abstract.
2019
Bredenkamp N, Yang J, Clarke J, Stirparo GG, von Meyenn F, Baker D, Drummond R, li D, Wu C, Rostovskaya M, et al (2019). Efficient RNA-mediated reprogramming of human somatic cells to naïve pluripotency facilitated by tankyrase inhibition.
Bredenkamp N, Stirparo GG, Nichols J, Smith A, Guo G (2019). The Cell-Surface Marker Sushi Containing Domain 2 Facilitates Establishment of Human Naive Pluripotent Stem Cells.
Stem Cell Reports,
12(6), 1212-1222.
Abstract:
The Cell-Surface Marker Sushi Containing Domain 2 Facilitates Establishment of Human Naive Pluripotent Stem Cells.
Recently naive human pluripotent stem cells (hPSCs) have been described that relate to an earlier stage of development than conventional hPSCs. Naive hPSCs remain challenging to generate and authenticate, however. Here we report that Sushi Containing Domain 2 (SUSD2) is a robust cell-surface marker of naive hPSCs in the embryo and in vitro. SUSD2 transcripts are enriched in the pre-implantation epiblast of human blastocysts and immunostaining shows localization of SUSD2 to KLF17-positive epiblast cells. SUSD2 mRNA is strongly expressed in naive hPSCs but is negligible in other hPSCs. SUSD2 immunostaining of live or fixed cells provides unambiguous discrimination of naive versus conventional hPSCs. SUSD2 staining or flow cytometry enable monitoring of naive hPSCs in maintenance culture, and their isolation and quantification during resetting of conventional hPSCs or somatic cell reprogramming. Thus SUSD2 is a powerful non-invasive tool for reliable identification and purification of the naive hPSC phenotype.
Abstract.
Author URL.
Bredenkamp N, Yang J, Clarke J, Stirparo GG, von Meyenn F, Dietmann S, Baker D, Drummond R, Ren Y, Li D, et al (2019). Wnt Inhibition Facilitates RNA-Mediated Reprogramming of Human Somatic Cells to Naive Pluripotency.
Stem Cell Reports,
13(6), 1083-1098.
Abstract:
Wnt Inhibition Facilitates RNA-Mediated Reprogramming of Human Somatic Cells to Naive Pluripotency.
In contrast to conventional human pluripotent stem cells (hPSCs) that are related to post-implantation embryo stages, naive hPSCs exhibit features of pre-implantation epiblast. Naive hPSCs are established by resetting conventional hPSCs, or are derived from dissociated embryo inner cell masses. Here we investigate conditions for transgene-free reprogramming of human somatic cells to naive pluripotency. We find that Wnt inhibition promotes RNA-mediated induction of naive pluripotency. We demonstrate application to independent human fibroblast cultures and endothelial progenitor cells. We show that induced naive hPSCs can be clonally expanded with a diploid karyotype and undergo somatic lineage differentiation following formative transition. Induced naive hPSC lines exhibit distinctive surface marker, transcriptome, and methylome properties of naive epiblast identity. This system for efficient, facile, and reliable induction of transgene-free naive hPSCs offers a robust platform, both for delineation of human reprogramming trajectories and for evaluating the attributes of isogenic naive versus conventional hPSCs.
Abstract.
Author URL.
2018
Guo G, von Meyenn F, Rostovskaya M, Clarke J, Dietmann S, Baker D, Sahakyan A, Myers S, Bertone P, Reik W, et al (2018). Epigenetic resetting of human pluripotency (vol 144, pg 2748, 2017).
DEVELOPMENT,
145(8).
Author URL.
Stirparo GG, Boroviak T, Guo G, Nichols J, Smith A, Bertone P (2018). Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human pre-implantation epiblast.
DEVELOPMENT,
145(3).
Author URL.
Stirparo GG, Boroviak T, Guo G, Nichols J, Smith A, Bertone P (2018). Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human pre-implantation epiblast (vol 145, dev158501, 2018).
DEVELOPMENT,
145(15).
Author URL.
Stirparo GG, Boroviak T, Guo G, Nichols J, Smith A, Bertone P (2018). Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human preimplantation epiblast.
DevelopmentAbstract:
Integrated analysis of single-cell embryo data yields a unified transcriptome signature for the human preimplantation epiblast
Single-cell profiling techniques create opportunities to delineate cell fate progression in mammalian development. Recent studies provide transcriptome data from human preimplantation embryos, in total comprising nearly 2000 individual cells. Interpretation of these data is confounded by biological factors such as variable embryo staging and cell-type ambiguity, as well as technical challenges in the collective analysis of datasets produced with different sample preparation and sequencing protocols. Here we address these issues to assemble a complete gene expression time course spanning human preimplantation embryogenesis. We identify key transcriptional features over developmental time and elucidate lineage-specific regulatory networks. We resolve post hoc cell-type assignment in the blastocyst, and define robust transcriptional prototypes that capture epiblast and primitive endoderm lineages. Examination of human pluripotent stem cell transcriptomes in this framework identifies culture conditions that sustain a naïve state pertaining to the inner cell mass. Our approach thus clarifies understanding both of lineage segregation in the early human embryo and of in vitro stem cell identity, and provides an analytical resource for comparative molecular embryology.
Abstract.
2017
Guo G, von Meyenn F, Rostovskaya M, Clarke J, Dietmann S, Baker D, Sahakyan A, Myers S, Bertone P, Reik W, et al (2017). Epigenetic resetting of human pluripotency.
DEVELOPMENT,
144(15), 2748-2763.
Author URL.
Guo G, von Meyenn F, Rostovskaya M, Clarke J, Dietmann S, Baker D, Sahakyan A, Myers S, Bertone P, Reik W, et al (2017). Epigenetic resetting of human pluripotency.
2016
Guo G, von Meyenn F, Santos F, Chen Y, Reik W, Bertone P, Smith A, Nichols J (2016). Naive Pluripotent Stem Cells Derived Directly from Isolated Cells of the Human Inner Cell Mass.
STEM CELL REPORTS,
6(4), 437-446.
Author URL.
2015
Takashima Y, Guo G, Loos R, Nichols J, Ficz G, Krueger F, Oxley D, Santos F, Clarke J, Mansfield W, et al (2015). Resetting Transcription Factor Control Circuitry toward Ground-State Pluripotency in Human (vol 158, pg 1254, 2014).
CELL,
162(2), 452-453.
Author URL.
2014
Takashima Y, Guo G, Loos R, Nichols J, Ficz G, Krueger F, Oxley D, Santos F, Clarke J, Mansfield W, et al (2014). Resetting Transcription Factor Control Circuitry toward Ground-State Pluripotency in Human.
CELL,
158(6), 1254-1269.
Author URL.
2012
Huang Y, Pettitt SJ, Guo G, Liu G, Li MA, Yang F, Bradley A (2012). Isolation of homozygous mutant mouse embryonic stem cells using a dual selection system.
NUCLEIC ACIDS RESEARCH,
40(3).
Author URL.
2011
Guo G, Huang Y, Humphreys P, Wang X, Smith A (2011). A PiggyBac-Based Recessive Screening Method to Identify Pluripotency Regulators.
PLOS ONE,
6(4).
Author URL.
Wang W, Yang J, Liu H, Lu D, Chen X, Zenonos Z, Campos LS, Rad R, Guo G, Zhang S, et al (2011). Rapid and efficient reprogramming of somatic cells to induced pluripotent stem cells by retinoic acid receptor gamma and liver receptor homolog 1.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,
108(45), 18283-18288.
Author URL.
2010
Guo G, Smith A (2010). A genome-wide screen in EpiSCs identifies Nr5a nuclear receptors as potent inducers of ground state pluripotency.
DEVELOPMENT,
137(19), 3185-3192.
Author URL.
Yang J, van Oosten AL, Theunissen TW, Guo G, Silva JCR, Smith A (2010). Stat3 Activation is Limiting for Reprogramming to Ground State Pluripotency.
CELL STEM CELL,
7(3), 319-328.
Author URL.
2009
Guo G, Yang J, Nichols J, Hall JS, Eyres I, Mansfield W, Smith A (2009). Klf4 reverts developmentally programmed restriction of ground state pluripotency.
DEVELOPMENT,
136(7), 1063-1069.
Author URL.
Silva J, Nichols J, Theunissen TW, Guo G, van Oosten AL, Barrandon O, Wray J, Yamanaka S, Chambers I, Smith A, et al (2009). Nanog is the Gateway to the Pluripotent Ground State.
CELL,
138(4), 722-737.
Author URL.
Hall J, Guo G, Wray J, Eyres I, Nichols J, Grotewold L, Morfopoulou S, Humphreys P, Mansfield W, Walker R, et al (2009). Oct4 and LIF/Stat3 Additively Induce Kruppel Factors to Sustain Embryonic Stem Cell Self-Renewal.
CELL STEM CELL,
5(6), 597-609.
Author URL.
2004
Guo G, Wang W, Bradley A (2004). Mismatch repair genes identified using genetic screens in Blm-deficient embryonic stem cells. Nature, 429(6994), 891-895.
