Close

Lactate Accelerates Mouse ES Cell Differentiation Towards the XEN Lineage

Mohamed I. Gatie, Tyler T. Cooper, Gilles A. Lajoie, Gregory M. Kelly

Preprint posted on 26 December 2020 https://www.biorxiv.org/content/10.1101/2020.12.14.422783v2.full

Enhance the in vitro conversion of embryonic stem cells to the XEN lineage with a single metabolite, lactate.

Selected by Nozomu Takata

Background

The yolk sac, a membranous structure situated outside of the embryo, has a variety of functions1. For instance, the yolk sac aids in nutrition and gas exchange between the mother and the developing embryo as well as the formation of the umbilical cord. Another important role is the production of embryonic blood cells, primitive macrophages and germ cells.

In amniotes, a major constituent of the yolk sac is created from cell derivatives of a particular cell lineage called the primitive endoderm (PrE), that appears as an epithelial monolayer visible shortly before implantation. It is one of the first three cell lineages, defined by complex mechanisms that remain elusive(Schematic the authors provided).

Due to the lack of accessibility to the PrE lineage on demand, there has been a plethora of well-established in vitro model systems allowing researchers to derive PrE cells and maintain them indefinitely in vitro as XEN cells3. XEN cells can be established from embryonic stem cells chemically4 or when the PrE-specific genes Gata4/65 or Sox176 are expressed in vitro. DAB2 is used as another XEN cell marker (Fig. S1), but it is likely other additional factors assist lineage specification toward the PrE.

Cells between preimplantation and post-implantation stages display distinct metabolic profiles7. The authors reported previously that F9 embryonal carcinoma cells, which are capable of adopting a XEN-like lineage, exhibited a metabolic phenotype reliant exclusively on glycolysis8. In addition, evidence indicates that the metabolic state between embryonic and extraembryonic lineages differs, suggesting that metabolism may be sufficient to influence lineage commitment. These phenomena have brought the authors to question why and how important glycolysis is for lineage specification.

In this preprint, the authors conducted metabolomic analysis on two distinct embryonic cell lineages and demonstrated that 1) XEN cells possess altered glycolytic enzyme levels involved in pyruvate and lactate homeostasis, 2) the cells are sensitive to glycolytic inhibition when compared with oxidative phosphorylation (OXPHOS) inhibition, 3) lactate is a key metabolite to enhance the XEN lineage.

 

Key findings

  1. Feeder-free XEN cells exhibit high sensitivity to glycolytic inhibition

The morphology of the ES cell colony was insensitive to treatment with the glucose blocker 2-DG or with oligomycin, an inhibitor of OXPHOS, while in XEN cells, 2-DG had a more pronounced effect not only on morphology but also on cell viability compared to oligomycin. It suggests that XEN cells were more sensitive to glycolytic inhibition than OXPHOS inhibition (Fig. 1)

  1. Metabolomic profiling between feeder-free ES and XEN cells identified lactate as a candidate metabolite

Next, the authors performed intracellular metabolomic analysis to identify changes in intracellular metabolite composition between ES and XEN cells. They revealed that 18 intracellular metabolites were significantly enriched in ES cells and 68 in XEN cells (Fig. 2), specifically, intracellular lactate levels were significantly higher in XEN cells. The data provided here indicate that XEN cells exhibit a unique metabolite profile distinct from the ES population.

  1. Elevated intracellular and extracellular lactate is observed in feeder-free XEN cells

LDHA converts the metabolite pyruvate into lactate while the reverse is mediated by LDHB. Therefore, the relative ratio of LDHA/LDHB could explain why lactate production was higher in XEN cells. Consistent with the finding from the metabolome analysis, LDHA expression was indeed notably higher in XEN cells than in ES cells in quantitative PCR and western blot analysis (Fig. 4).

How about the alternative possibility that pyruvate in XEN cells travels into mitochondria and facilitates OXPHOS to generate energy? Expression analysis of the mitochondrial pyruvate carrier Mpc1 showed significantly reduced expression in XEN cells. Therefore, pyruvate in XEN cells would likely fail to enter the mitochondria and instead, is preferentially converted into lactate by the enhanced LDHA activity.

The authors conducted functional tests to prove the hypothesis that lactate can facilitate cell fate. To promote intracellular lactate accumulation, the cells were incubated with L-lactate or AZD3965, a potent inhibitor to block lactate shuttling to the extracellular space. Intriguingly, both treatments enhanced Gata6 and Dab2 expression. Therefore, promoting intracellular lactate accumulation enhanced XEN induction in vitro, highlighting a novel role for lactate as a signaling molecule involved in the differentiation of XEN cells. These results suggest that lactate participates in specifying ES cells towards the XEN lineage.

In summary, the findings strongly suggest that feeder-free XEN cells are sensitive to glycolytic inhibition, produce high levels of lactate, which enhances XEN induction in vitro. This study revealed a functional association between metabolism and cell lineage specification.

 

Why I like to highlight this preprint

I like the questions the authors raised and how they were challenged in this study.

There are a growing number of reports suggesting that metabolites play unexpected roles in transcriptional changes following epigenetic modifications. In multicellular systems, it would not be so surprising that layers of multiple signalling cascades secure and regulate a variety of cellular activities that lead to changes in gene expression. However, the contribution of metabolites and key mechanisms remain less understood.

Embryonic development is a beautiful system that exists before birth and one of the best platforms to witness cell lineage specification. The earlier the developmental stage in vivo, the more difficult it generally is to get access to a small number of cell types. Therefore, this study uses an in vitro culture system that clearly distinguishes two key cell types and overcomes a quantitative hurdle when metabolomics is performed.

The discovery of glycolysis and its product lactate in the XEN cells may open the avenue to not only better understand the new aspect of metabolism in mammalian cell specification, but perhaps give us better treatments for complications during pregnancy in the future 9,10.

 

Questions to the authors

  • Is this mechanism happening in real embryos? If so, what could be the trigger for the sensitivity towards glycolysis? It would be nice if the authors could validate the findings in vivo in the future.

 

  • What happens when LDHA is inhibited or functionally KO? I assume lactate production will be reduced. Would this treatment stop the maintenance of XEN cell fate?

 

  • Lactate accelerates XEN cell markers. Is there any clear change in epigenetic modification? And if so, how would that be caused?

 

  • It wouldn’t be surprising if other embryonic cell types also prefer to consume lactate. Testing other lineages would be of great interest to reveal a general mechanism during embryonic development.

 

  • I am curious about the dynamics of glycolysis activity, which could be analyzed by visualizing the final product lactate in live by a fluorescent reporter system11.

 

 References

1          Donovan, M. F. & Bordoni, B. in StatPearls     (2021).

2          Hermitte, S. & Chazaud, C. Primitive endoderm differentiation: from specification to epithelium formation. Philos Trans R Soc Lond B Biol Sci 369, doi:10.1098/rstb.2013.0537 (2014).

3          Niakan, K. K., Schrode, N., Cho, L. T. & Hadjantonakis, A. K. Derivation of extraembryonic endoderm stem (XEN) cells from mouse embryos and embryonic stem cells. Nat Protoc 8, 1028-1041, doi:10.1038/nprot.2013.049 (2013).

4          Anderson, K. G. V. et al. Insulin fine-tunes self-renewal pathways governing naive pluripotency and extra-embryonic endoderm. Nat Cell Biol 19, 1164-1177, doi:10.1038/ncb3617 (2017).

5          Fujikura, J. et al. Differentiation of embryonic stem cells is induced by GATA factors. Gene Dev 16, 784-789, doi:DOI 10.1101/gad.968802 (2002).

6          McDonald, A. C., Biechele, S., Rossant, J. & Stanford, W. L. Sox17-mediated XEN cell conversion identifies dynamic networks controlling cell-fate decisions in embryo-derived stem cells. Cell Rep 9, 780-793, doi:10.1016/j.celrep.2014.09.026 (2014).

7          Zhou, W. et al. HIF1alpha induced switch from bivalent to exclusively glycolytic metabolism during ESC-to-EpiSC/hESC transition. EMBO J 31, 2103-2116, doi:10.1038/emboj.2012.71 (2012).

8          Gatie, M. I. & Kelly, G. M. Metabolic profile and differentiation potential of extraembryonic endoderm-like cells. cell Death Discov 4, 42, doi:10.1038/s41420-018-0102-1 (2018).

9          Moradan, S. & Forouzeshfar, M. Are abnormal yolk sac characteristics important factors in abortion rates? Int J Fertil Steril 6, 127-130 (2012).

10        Cho, F. N., Chen, S. N., Tai, M. H. & Yang, T. L. The quality and size of yolk sac in early pregnancy loss. Aust N Z J Obstet Gynaecol 46, 413-418, doi:10.1111/j.1479-828X.2006.00627.x (2006).

11        San Martin, A. et al. A genetically encoded FRET lactate sensor and its use to detect the Warburg effect in single cancer cells. PLoS One 8, e57712, doi:10.1371/journal.pone.0057712 (2013).

 

Tags: cell lineage, metabolomic profile

Posted on: 7 April 2021

doi: https://doi.org/10.1242/prelights.28089

Read preprint (No Ratings Yet)

Have your say

Your email address will not be published. Required fields are marked *

This site uses Akismet to reduce spam. Learn how your comment data is processed.

Sign up to customise the site to your preferences and to receive alerts

Register here

preLists in the cell biology category:

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

preLights peer support – preprints of interest

This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.

 



List by preLights peer support

The Society for Developmental Biology 82nd Annual Meeting

This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.

 



List by Joyce Yu, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

Planar Cell Polarity – PCP

This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.

 



List by Ana Dorrego-Rivas

BioMalPar XVI: Biology and Pathology of the Malaria Parasite

[under construction] Preprints presented at the (fully virtual) EMBL BioMalPar XVI, 17-18 May 2020 #emblmalaria

 



List by Dey Lab, Samantha Seah

1

Cell Polarity

Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

 



List by Yamini Ravichandran

TAGC 2020

Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

 



List by Maiko Kitaoka et al.

3D Gastruloids

A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.

 



List by Paul Gerald L. Sanchez and Stefano Vianello

ECFG15 – Fungal biology

Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome

 



List by Hiral Shah

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

EMBL Seeing is Believing – Imaging the Molecular Processes of Life

Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

 



List by Dey Lab

Autophagy

Preprints on autophagy and lysosomal degradation and its role in neurodegeneration and disease. Includes molecular mechanisms, upstream signalling and regulation as well as studies on pharmaceutical interventions to upregulate the process.

 



List by Sandra Malmgren Hill

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Cellular metabolism

A curated list of preprints related to cellular metabolism at Biorxiv by Pablo Ranea Robles from the Prelights community. Special interest on lipid metabolism, peroxisomes and mitochondria.

 



List by Pablo Ranea Robles

BSCB/BSDB Annual Meeting 2019

Preprints presented at the BSCB/BSDB Annual Meeting 2019

 



List by Dey Lab

MitoList

This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.

 



List by Sandra Franco Iborra

Biophysical Society Annual Meeting 2019

Few of the preprints that were discussed in the recent BPS annual meeting at Baltimore, USA

 



List by Joseph Jose Thottacherry

ASCB/EMBO Annual Meeting 2018

This list relates to preprints that were discussed at the recent ASCB conference.

 



List by Dey Lab, Amanda Haage

Also in the developmental biology category:

GfE/ DSDB meeting 2024

This preList highlights the preprints discussed at the 2024 joint German and Dutch developmental biology societies meeting that took place in March 2024 in Osnabrück, Germany.

 



List by Joyce Yu

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

preLights peer support – preprints of interest

This is a preprint repository to organise the preprints and preLights covered through the 'preLights peer support' initiative.

 



List by preLights peer support

The Society for Developmental Biology 82nd Annual Meeting

This preList is made up of the preprints discussed during the Society for Developmental Biology 82nd Annual Meeting that took place in Chicago in July 2023.

 



List by Joyce Yu, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

Journal of Cell Science meeting ‘Imaging Cell Dynamics’

This preList highlights the preprints discussed at the JCS meeting 'Imaging Cell Dynamics'. The meeting was held from 14 - 17 May 2023 in Lisbon, Portugal and was organised by Erika Holzbaur, Jennifer Lippincott-Schwartz, Rob Parton and Michael Way.

 



List by Helen Zenner

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

2nd Conference of the Visegrád Group Society for Developmental Biology

Preprints from the 2nd Conference of the Visegrád Group Society for Developmental Biology (2-5 September, 2021, Szeged, Hungary)

 



List by Nándor Lipták

Fibroblasts

The advances in fibroblast biology preList explores the recent discoveries and preprints of the fibroblast world. Get ready to immerse yourself with this list created for fibroblasts aficionados and lovers, and beyond. Here, my goal is to include preprints of fibroblast biology, heterogeneity, fate, extracellular matrix, behavior, topography, single-cell atlases, spatial transcriptomics, and their matrix!

 



List by Osvaldo Contreras

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

EMBL Conference: From functional genomics to systems biology

Preprints presented at the virtual EMBL conference "from functional genomics and systems biology", 16-19 November 2020

 



List by Jesus Victorino

Single Cell Biology 2020

A list of preprints mentioned at the Wellcome Genome Campus Single Cell Biology 2020 meeting.

 



List by Alex Eve

Society for Developmental Biology 79th Annual Meeting

Preprints at SDB 2020

 



List by Irepan Salvador-Martinez, Martin Estermann

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

Planar Cell Polarity – PCP

This preList contains preprints about the latest findings on Planar Cell Polarity (PCP) in various model organisms at the molecular, cellular and tissue levels.

 



List by Ana Dorrego-Rivas

Cell Polarity

Recent research from the field of cell polarity is summarized in this list of preprints. It comprises of studies focusing on various forms of cell polarity ranging from epithelial polarity, planar cell polarity to front-to-rear polarity.

 



List by Yamini Ravichandran

TAGC 2020

Preprints recently presented at the virtual Allied Genetics Conference, April 22-26, 2020. #TAGC20

 



List by Maiko Kitaoka et al.

3D Gastruloids

A curated list of preprints related to Gastruloids (in vitro models of early development obtained by 3D aggregation of embryonic cells). Updated until July 2021.

 



List by Paul Gerald L. Sanchez and Stefano Vianello

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

EDBC Alicante 2019

Preprints presented at the European Developmental Biology Congress (EDBC) in Alicante, October 23-26 2019.

 



List by Sergio Menchero et al.

EMBL Seeing is Believing – Imaging the Molecular Processes of Life

Preprints discussed at the 2019 edition of Seeing is Believing, at EMBL Heidelberg from the 9th-12th October 2019

 



List by Dey Lab

SDB 78th Annual Meeting 2019

A curation of the preprints presented at the SDB meeting in Boston, July 26-30 2019. The preList will be updated throughout the duration of the meeting.

 



List by Alex Eve

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

Young Embryologist Network Conference 2019

Preprints presented at the Young Embryologist Network 2019 conference, 13 May, The Francis Crick Institute, London

 



List by Alex Eve

Pattern formation during development

The aim of this preList is to integrate results about the mechanisms that govern patterning during development, from genes implicated in the processes to theoritical models of pattern formation in nature.

 



List by Alexa Sadier

BSCB/BSDB Annual Meeting 2019

Preprints presented at the BSCB/BSDB Annual Meeting 2019

 



List by Dey Lab

Zebrafish immunology

A compilation of cutting-edge research that uses the zebrafish as a model system to elucidate novel immunological mechanisms in health and disease.

 



List by Shikha Nayar

Also in the molecular biology category:

‘In preprints’ from Development 2022-2023

A list of the preprints featured in Development's 'In preprints' articles between 2022-2023

 



List by Alex Eve, Katherine Brown

CSHL 87th Symposium: Stem Cells

Preprints mentioned by speakers at the #CSHLsymp23

 



List by Alex Eve

9th International Symposium on the Biology of Vertebrate Sex Determination

This preList contains preprints discussed during the 9th International Symposium on the Biology of Vertebrate Sex Determination. This conference was held in Kona, Hawaii from April 17th to 21st 2023.

 



List by Martin Estermann

Alumni picks – preLights 5th Birthday

This preList contains preprints that were picked and highlighted by preLights Alumni - an initiative that was set up to mark preLights 5th birthday. More entries will follow throughout February and March 2023.

 



List by Sergio Menchero et al.

CellBio 2022 – An ASCB/EMBO Meeting

This preLists features preprints that were discussed and presented during the CellBio 2022 meeting in Washington, DC in December 2022.

 



List by Nadja Hümpfer et al.

EMBL Synthetic Morphogenesis: From Gene Circuits to Tissue Architecture (2021)

A list of preprints mentioned at the #EESmorphoG virtual meeting in 2021.

 



List by Alex Eve

FENS 2020

A collection of preprints presented during the virtual meeting of the Federation of European Neuroscience Societies (FENS) in 2020

 



List by Ana Dorrego-Rivas

ECFG15 – Fungal biology

Preprints presented at 15th European Conference on Fungal Genetics 17-20 February 2020 Rome

 



List by Hiral Shah

ASCB EMBO Annual Meeting 2019

A collection of preprints presented at the 2019 ASCB EMBO Meeting in Washington, DC (December 7-11)

 



List by Madhuja Samaddar et al.

Lung Disease and Regeneration

This preprint list compiles highlights from the field of lung biology.

 



List by Rob Hynds

MitoList

This list of preprints is focused on work expanding our knowledge on mitochondria in any organism, tissue or cell type, from the normal biology to the pathology.

 



List by Sandra Franco Iborra
Close