A single cell characterisation of human embryogenesis identifies pluripotency transitions and putative anterior hypoblast centre

Molè, Matteo A.; Coorens, Tim; Shahbazi, Marta; Weberling, Antonia; Weatherbee, Bailey A. T.; Gantner, Carlos W.; Sancho-Serra, Carmen; Richardson, Lucy; Drinkwater, Abbie; Syed, Najma; Engley, Stephanie; Snell, Philip; Christie, Leila; Elder, Kay; Campbell, Alison; Fishel, Simon; Behjati, Sam; Vento‐Tormo, Roser; Zernicka‐Goetz, Magdalena

doi:10.1038/s41467-021-23758-w

Cited by 84 publications

(101 citation statements)

References 77 publications

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“…Consistent with the staining pattern and in line with a previous report, we observed a steady reduction of OCT4 transcripts from day 7 to day 12, while the abundance of NANOG and SOX2 transcripts did not change between day 7 and day 9 and eventually decreased by day 12 29 . Similar to human pregastrulation development, we found a decreasing abundance of transcripts associated with naïve pluripotency ( KLF4 , KLF17 , PRDM8 , TFCP2L1 , ZFP42 , UTF1 ), while markers for primed pluripotency, that increased in human ( FGF2 , DNMT3B , SOX11 , SFRP2 , SALL2 ), 17 did not change from day 7 until day 12. Van Leeuwen et al 13 detected NODAL transcripts in the EPI of Rauber's layer (RL) stage (day 10–11) embryos and suggested NODAL activation through the convertase FURIN, which they detected in the RL.…”

Section: Discussionsupporting

confidence: 56%

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

et al. 2022

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The mammalian blastocyst undergoes two lineage segregations, that is, formation of the trophectoderm and subsequently differentiation of the hypoblast (HB) from the inner cell mass, leaving the epiblast (EPI) as the remaining pluripotent lineage. To clarify the expression patterns of markers specific for these lineages in bovine embryos, we analyzed day 7, 9, and 12 blastocysts completely produced in vivo by staining for OCT4, NANOG, SOX2 (EPI), and GATA6, SOX17 (HB) and identified genes specific for these developmental stages in a global transcriptomics approach. To study the role of OCT4, we generated OCT4‐deficient (OCT4 KO) embryos via somatic cell nuclear transfer or in vitro fertilization. OCT4 KO embryos reached the expanded blastocyst stage by day 8 but lost NANOG and SOX17 expression, while SOX2 and GATA6 were unaffected. Blastocysts transferred to recipient cows from day 6 to 9 expanded, but the OCT4 KO phenotype was not rescued by the uterine environment. Exposure of OCT4 KO embryos to exogenous FGF4 or chimeric complementation with OCT4 intact embryos did not restore NANOG or SOX17 in OCT4‐deficient cells. Our data show that OCT4 is required cell autonomously for the maintenance of pluripotency of the EPI and differentiation of the HB in bovine embryos.

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Section: Discussionsupporting

confidence: 56%

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

et al. 2022

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“…On the other hand, we found a significant reduction in total cell numbers, which is in contrast to mouse Nanog -KO embryos, where the loss of NANOG does not impede cell proliferation until the E3.5 blastocyst stage [ 1 ]. Interestingly, the reduced total cell number in NANOG -KO embryos reached normal levels when the embryos were cultivated with exogenous FGF4, where the proliferative impact of FGF4 [ 17 , 37 , 38 , 39 ] evidently alleviated the reduction of total cell numbers in the absence of NANOG. We hypothesize that in NANOG -KO embryos, the absence of FGF4 expressing EPI precursor cells causes the reduced cell number.…”

Section: Discussionmentioning

confidence: 99%

Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

Springer

Zakhartchenko

Wolf

et al. 2021

Cells

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The role of the pluripotency factor NANOG during the second embryonic lineage differentiation has been studied extensively in mouse, although species-specific differences exist. To elucidate the role of NANOG in an alternative model organism, we knocked out NANOG in fibroblast cells and produced bovine NANOG-knockout (KO) embryos via somatic cell nuclear transfer (SCNT). At day 8, NANOG-KO blastocysts showed a decreased total cell number when compared to controls from SCNT (NT Ctrl). The pluripotency factors OCT4 and SOX2 as well as the hypoblast (HB) marker GATA6 were co-expressed in all cells of the inner cell mass (ICM) and, in contrast to mouse Nanog-KO, expression of the late HB marker SOX17 was still present. We blocked the MEK-pathway with a MEK 1/2 inhibitor, and control embryos showed an increase in NANOG positive cells, but SOX17 expressing HB precursor cells were still present. NANOG-KO together with MEK-inhibition was lethal before blastocyst stage, similarly to findings in mouse. Supplementation of exogenous FGF4 to NANOG-KO embryos did not change SOX17 expression in the ICM, unlike mouse Nanog-KO embryos, where missing SOX17 expression was completely rescued by FGF4. We conclude that NANOG mediated FGF/MEK signaling is not required for HB formation in the bovine embryo and that another—so far unknown—pathway regulates HB differentiation.

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“…In Figure 1 , I revisit the question of whether the enrichment of X-linked genes we identified in ASD cord blood, a tissue derived from the epiblast lineage, could be related to their variable cellular expression and XCI status at a stage closer to post-implantation in humans. For this, the transcripts XACT , XIST , and two ASD relevant genes from Table 1 , PTCHD1 and MID1 , were plotted from a recent single cell RNAseq data set from 16 human embryos cultured to 9 or 11 days post fertilization, bioinformatically sorted into epiblast, hypoblast, syncytiotrophoblast, and cytotrophoblast ( www.humanembryo.org ) ( Mole et al, 2021 ). Interestingly, among the cell type markers of maturity in the epiblast identified in this study were transcriptional changes to two genes encoding DNA methyltransferases, specifically a decrease in DNMT3L and an increase in DNMT3B (25).…”

Section: Introductionmentioning

confidence: 99%

“…Top row shows drawings of pre- (day 6 post fertilization) and peri-implantation human embryos with the major cell lineages labelled (from BioRender.com ). Bottom panel presents data acquired from Mole et al, 2021 dataset in www.humanembryo.org which includes single cell RNA-seq from 16 human peri-implantation embryos, 8 at day 9 and 8 at day 11. While sex of embryos was not determined in the published study, a negligible level of SRY expression was observed, consistent with female sex.…”

Section: Introductionmentioning

confidence: 99%

X Chromosome Inactivation Timing is Not eXACT: Implications for Autism Spectrum Disorders

LaSalle

2022

Front. Genet.

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The etiology of autism spectrum disorders (ASD) is complex, involving different combinations of genetic and environmental factors. My lab’s approach has been to investigate DNA methylation as a tractable genome-wide modification at the interface of these complex interactions, reflecting past and future events in the molecular pathogenesis of ASD. Since X-linked genes were enriched in DNA methylation differences discovered from cord blood from newborns later diagnosed with ASD, this has prompted me to review and revisit the recent advancements in the field of X chromosome inactivation (XCI), particularly in humans and other primates. In this Perspective, I compare XCI mechanisms in different mammalian species, including the finding of the noncoding transcript XACT associated with X chromosome erosion in human pluripotent stem cells and recent findings from non-human primate post-implantation embryos. I focus on the experimentally challenging peri- and post-implantation stages of human development when the timing of XCI is prolonged and imprecise in humans. Collectively, this research has raised some important unanswered questions involving biased sex ratios in human births and the male bias in the incidence of ASD.

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A single cell characterisation of human embryogenesis identifies pluripotency transitions and putative anterior hypoblast centre

Cited by 84 publications

References 77 publications

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

OCT4/POU5F1 is indispensable for the lineage differentiation of the inner cell mass in bovine embryos

Hypoblast Formation in Bovine Embryos Does Not Depend on NANOG

X Chromosome Inactivation Timing is Not eXACT: Implications for Autism Spectrum Disorders

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