A regulatory circuitry comprised of miR-302 and the transcription factors OCT4 and NR2F2 regulates human embryonic stem cell differentiation
Open accessAbstract
Multiple levels of control are in play to regulate pluripotency and differentiation in human embryonic stem cells (hESCs). At the transcriptional level, the core factors OCT4, NANOG and SOX2 form a positive autoregulatory loop that is pivotal for maintaining the undifferentiated state. At the post-transcriptional level, microRNAs (miRNAs) belonging to the miR-302 family are emerging as key players in the control of proliferation and cell fate determination during differentiation. Here, we show that the transcriptional factors OCT4 and NR2F2 (COUP-TFII) and the miRNA miR-302 are linked in a regulatory circuitry that critically regulate both pluripotency and differentiation in hESCs. In the undifferentiated state, both OCT4 and the OCT4-induced miR-302 directly repress NR2F2 at the transcriptional and post-transcriptional level, respectively. Conversely, NR2F2 directly inhibits OCT4 during differentiation, triggering a positive feedback loop for its own expression. In addition, we show that regulation of NR2F2 activity itself relies on alternative splicing and transcriptional start site choice to generate a full-length transcriptionally active isoform and shorter variants, which enhance the activity of the long isoform. During hESC differentiation, NR2F2 is first detected at the earliest steps of neural induction and thus is among the earliest human embryonic neural markers. Finally, our functional analysis points to a crucial role for NR2F2 in the activation of neural genes during early differentiation in humans. These findings introduce a new molecular player in the context of early embryonic stem cell state and cell fate determination in humans.
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NR2F2 is involved in regulating embryonic stem cell differentiation and implicated in neural crest development, and has been under selection in the domesticated fox.
“The functionally related nuclear receptor NR2F2 is involved in regulating embryonic stem cell differentiation [66] and implicated in neural crest development, and has been under selection in the domesticated fox [67].”
From Self-domestication in Homo sapiens: Insights from comparative genomics (2017) · cites this paper as [66]
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Cites
MicroRNAs: Target Recognition and Regulatory Functions
Bartel, D. P. (2009) · Cell · doi:10.1016/j.cell.2009.01.002 · Cited by 20,068 · see citing works
Core Transcriptional Regulatory Circuitry in Human Embryonic Stem Cells
Boyer, L. A., Lee, T. I., Cole, M. F., Johnstone, S. E., Levine, S. S., Zucker, J. P., Guenther, M. G., Kumar, R. M., Murray, H. L., Jenner, R. G., et al. (2005) · Cell · doi:10.1016/j.cell.2005.08.020 · Cited by 4,505 · see citing works
A Mammalian microRNA Expression Atlas Based on Small RNA Library Sequencing
Landgraf, P., Rusu, M., Sheridan, R., Sewer, A., Iovino, N., Aravin, A., Pfeffer, S., Rice, A., Kamphorst, A. O., Landthaler, M., et al. (2007) · Cell · doi:10.1016/j.cell.2007.04.040 · Cited by 3,626 · see citing works
Nichols, J., Zevnik, B., Anastassiadis, K., Niwa, H., Klewe-Nebenius, D., Chambers, I., Schöler, H. R., & Smith, A. G. (1998) · Cell · doi:10.1016/s0092-8674(00)81769-9 · Cited by 3,394 · see citing works
Functional Expression Cloning of Nanog, a Pluripotency Sustaining Factor in Embryonic Stem Cells
Chambers, I., Colby, D., Robertson, M., Nichols, J., Lee, S., Tweedie, S., & Smith, A. G. (2003) · Cell · doi:10.1016/s0092-8674(03)00392-1 · Cited by 3,255 · see citing works
Control of Developmental Regulators by Polycomb in Human Embryonic Stem Cells
Lee, T. I., Jenner, R. G., Boyer, L. A., Guenther, M. G., Levine, S. S., Kumar, R., Chevalier, B. S., Johnstone, S. E., Cole, M. F., Isono, K., et al. (2006) · Cell · doi:10.1016/j.cell.2006.02.043 · Cited by 2,238 · see citing works
Dicer is essential for mouse development
Bernstein, E., Kim, S. Y., Carmell, M., Murchison, E., Alcorn, H. L., Li, M. Z., Mills, A., Elledge, S., Anderson, K., & Hannon, G. (2003) · Nature Genetics · doi:10.1038/ng1253 · Cited by 2,100 · see citing works
Connecting microRNA Genes to the Core Transcriptional Regulatory Circuitry of Embryonic Stem Cells
Marson, A., Levine, S. S., Cole, M. F., Frampton, G. M., Brambrink, T., Johnstone, S. E., Guenther, M. G., Johnston, W. K., Wernig, M., Newman, J. J., et al. (2008) · Cell · doi:10.1016/j.cell.2008.07.020 · Cited by 1,361 · see citing works
Kanellopoulou, C., Muljo, S. A., Kung, A. L., Ganesan, S., Drapkin, R., Jenuwein, T., Livingston, D. M., & Rajewsky, K. (2005) · Genes & Development · doi:10.1101/gad.1248505 · Cited by 1,256 · see citing works
Stark, A., Brennecke, J., Bushati, N., Russell, R. B., & Cohen, S. M. (2005) · Cell · doi:10.1016/j.cell.2005.11.023 · Cited by 1,080 · see citing works
DGCR8 is essential for microRNA biogenesis and silencing of embryonic stem cell self-renewal
Wang, Y., Medvid, R., Melton, C. A., Jaenisch, R., & Blelloch, R. H. (2007) · Nature Genetics · doi:10.1038/ng1969 · Cited by 1,026 · see citing works
Fazi, F., Rosa, A., Fatica, A., Gelmetti, V., Marchis, M. L. D., Nervi, C., & Bozzoni, I. (2005) · Cell · doi:10.1016/j.cell.2005.09.023 · Cited by 984 · see citing works
Self-citationHuman embryonic stem cells express a unique set of microRNAs
Suh, M. R., Lee, Y., Kim, J. Y., Kim, S.-K., Moon, S., Lee, J. Y., Cha, K., Chung, H., Yoon, H. S., Moon, S. Y., et al. (2004) · Developmental Biology · doi:10.1016/j.ydbio.2004.02.019 · Cited by 974 · see citing works
Morin, R. D., O’Connor, M. D., Griffith, M., Kuchenbauer, F., Delaney, A., Prabhu, A.-L., Zhao, Y., McDonald, H., Zeng, T., Hirst, M., et al. (2008) · Genome Research · doi:10.1101/gr.7179508 · Cited by 972 · see citing works
Yuan, H., Corbi, N., Basilico, C., & Dailey, L. (1995) · Genes & Development · doi:10.1101/gad.9.21.2635 · Cited by 811 · see citing works
Developmental Defects in Mouse Embryos Lacking N-Cadherin
Radice, G. L., Rayburn, H., Matsunami, H., Knudsen, K. A., Takeichi, M., & Hynes, R. O. (1997) · Developmental Biology · doi:10.1006/dbio.1996.8443 · Cited by 729 · see citing works
Oliver, G. C., Mailhos, A., Wehr, R., Copeland, N. G., Jenkins, N. A., & Gruß, P. (1995) · Development · doi:10.1242/dev.121.12.4045 · Cited by 674 · see citing works
Chew, J.-L., Loh, Y., Zhang, W., Chen, X., Tam, W. L., Yeap, L.-S., Li, P., Ang, Y.-S., Lim, B., Robson, P., et al. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.25.14.6031-6046.2005 · Cited by 641 · see citing works
Bernstein, E., Duncan, E. M., Masui, O., Gil, J., Heard, É., & Allis, C. D. (2006) · Molecular and Cellular Biology · doi:10.1128/mcb.26.7.2560-2569.2006 · Cited by 531 · see citing works
The orphan nuclear receptor COUP-TFII is required for angiogenesis and heart developmentPreprint
Pereira, F. A., Qiu, Y., Zhou, G., Tsai, M., & Tsai, S. Y. (1999) · Genes & Development · doi:10.1101/gad.13.8.1037 · Cited by 526 · see citing works
Canalization of development by microRNAs
Hornstein, E., & Shomron, N. (2006) · Nature Genetics · doi:10.1038/ng1803 · Cited by 511 · see citing works
Oct4/Sox2-Regulated miR-302 Targets Cyclin D1 in Human Embryonic Stem Cells
Card, D. A. G., Hebbar, P. B., Li, L., Trotter, K. W., Komatsu, Y., Mishina, Y. M., & Archer, T. (2008) · Molecular and Cellular Biology · doi:10.1128/mcb.00359-08 · Cited by 493 · see citing works
Lhx2, a LIM homeobox gene, is required for eye, forebrain, and definitive erythrocyte development
Porter, F. D., Drago, J., Xu, Y., Cheema, S. S., Wassif, C. A., Huang, S.-P., Lee, E., Grinberg, A., Massalas, J. S., Bodine, D. M., et al. (1997) · Development · doi:10.1242/dev.124.15.2935 · Cited by 491 · see citing works
A MicroRNA Imparts Robustness against Environmental Fluctuation during Development
Li, X., Cassidy, J. J., Reinke, C., Fischboeck, S., & Carthew, R. W. (2009) · Cell · doi:10.1016/j.cell.2009.01.058 · Cited by 445 · see citing works
Epigenetic signatures of stem-cell identity
Spivakov, M., & Fisher, A. G. (2007) · Nature Reviews Genetics · doi:10.1038/nrg2046 · Cited by 382 · see citing works
Hay, D. C., Sutherland, L., Clark, A., & Burdon, T. (2004) · Stem Cells · doi:10.1634/stemcells.22-2-225 · Cited by 376 · see citing works
Vallier, L., Reynolds, D. S., & Pedersen, R. A. (2004) · Developmental Biology · doi:10.1016/j.ydbio.2004.08.031 · Cited by 350 · see citing works
Chick Ovalbumin Upstream Promoter-Transcription Factors (COUP-TFs): Coming of Age
Tsai, S. Y., & Tsai, M. (1997) · Endocrine Reviews · doi:10.1210/edrv.18.2.0294 · Cited by 334 · see citing works
Nagai, T., Aruga, J., Takada, S., Günther, T., Spörle, R., Schughart, K., & Mikoshiba, K. (1997) · Developmental Biology · doi:10.1006/dbio.1996.8449 · Cited by 318 · see citing works
Pankratz, M. T., Li, X.-J., LaVaute, T. M., Lyons, E. A., Chen, X., & Zhang, S. (2007) · Stem Cells · doi:10.1634/stemcells.2006-0707 · Cited by 315 · see citing works
Bar, M., Wyman, S. K., Fritz, B. R., Qi, J., Garg, K. S., Parkin, R. K., Kroh, E. M., Bendoraite, A., Mitchell, P. S., Nelson, A. M., et al. (2008) · Stem Cells · doi:10.1634/stemcells.2008-0356 · Cited by 300 · see citing works
Smith, J. R., Vallier, L., Lupo, G., Alexander, M., Harris, W. A., & Pedersen, R. A. (2007) · Developmental Biology · doi:10.1016/j.ydbio.2007.10.003 · Cited by 295 · see citing works
High‐Efficiency RNA Interference in Human Embryonic Stem Cells
Zaehres, H., Lensch, M. W., Daheron, L. M., Stewart, S. A., Itskovitz‐Eldor, J., & Daley, G. Q. (2005) · Stem Cells · doi:10.1634/stemcells.2004-0252 · Cited by 268 · see citing works
Naka, H., Nakamura, S., Shimazaki, T., & Okano, H. (2008) · Nature Neuroscience · doi:10.1038/nn.2168 · Cited by 243 · see citing works
Qiu, Y., Pereira, F. A., DeMayo, F. J., Lydon, J. P., Tsai, S. Y., & Tsai, M. (1997) · Genes & Development · doi:10.1101/gad.11.15.1925 · Cited by 215 · see citing works
Jesus, A. B.-D., Romero‐López, C., Lucena-Aguilar, G., Melen, G. J., Sánchez, L., Ligero, G., Berzal‐Herranz, A., & Menéndez, P. R. (2008) · Molecular and Cellular Biology · doi:10.1128/mcb.00398-08 · Cited by 214 · see citing works
Identification of COUP-TFII Orphan Nuclear Receptor as a Retinoic Acid–Activated Receptor
Kruse, S. W., Suino-Powell, K. M., Zhou, X. E., Kretschman, J. E., Reynolds, R. A., Vonrhein, C., Xu, Y., Wang, L., Tsai, S. Y., Tsai, M., et al. (2008) · PLoS Biology · doi:10.1371/journal.pbio.0060227 · Cited by 208 · see citing works
Rosa, A., Spagnoli, F. M., & Brivanlou, A. H. (2009) · Developmental Cell · doi:10.1016/j.devcel.2009.02.007 · Cited by 206 · see citing works
Self-citationLacoste, A., Berenshteyn, F., & Brivanlou, A. H. (2009) · Cell stem cell · doi:10.1016/j.stem.2009.07.011 · Cited by 202 · see citing works
Self-citationCOUP-TF orphan nuclear receptors in development and differentiation
Pereira, F. A., Tsai, M., & Tsai, S. Y. (2000) · Cellular and Molecular Life Sciences · doi:10.1007/pl00000624 · Cited by 193 · see citing works
Gu, P., Menuet, D. L., Chung, A. C.-K., Mancini, M. A., Wheeler, D. A., & Cooney, A. J. (2005) · Molecular and Cellular Biology · doi:10.1128/mcb.25.19.8507-8519.2005 · Cited by 186 · see citing works
Zhou, C. J., Qiu, Y., Pereira, F. A., Crair, M. C., Tsai, S. Y., & Tsai, M. (1999) · Neuron · doi:10.1016/s0896-6273(00)81032-6 · Cited by 185 · see citing works
COUP-TFI: an intrinsic factor for early regionalization of the neocortexPreprint
Zhou, C. J., Tsai, S. Y., & Tsai, M. (2001) · Genes & Development · doi:10.1101/gad.913601 · Cited by 184 · see citing works
The miR-302-367 cluster as a potential stemness regulator in ESCs
Jesus, A. B.-D., Lucena-Aguilar, G., & Menéndez, P. R. (2009) · Cell Cycle · doi:10.4161/cc.8.3.7554 · Cited by 165 · see citing works
Qiu, Y., Cooney, A. J., Kuratani, S., DeMayo, F. J., Tsai, S. Y., & Tsai, M. (1994) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.91.10.4451 · Cited by 161 · see citing works
Contribution of human embryonic stem cells to mouse blastocysts
James, D., Noggle, S., Swigut, T., & Brivanlou, A. H. (2006) · Developmental Biology · doi:10.1016/j.ydbio.2006.03.026 · Cited by 155 · see citing works
Self-citationBen-Shushan, E., Sharir, H., Pikarsky, E., & Bergman, Y. (1995) · Molecular and Cellular Biology · doi:10.1128/mcb.15.2.1034 · Cited by 153 · see citing works
Chick Ovalbumin Upstream Promoter-Transcription Factors (COUP-TFs): Coming of Age
Tsai, S. Y. (1997) · Endocrine Reviews · doi:10.1210/er.18.2.229 · Cited by 133 · see citing works
The COUP-TF nuclear receptors regulate cell migration in the mammalian basal forebrain
Tripodi, M., Filosa, A., Armentano, M., & Studer, M. (2004) · Development · doi:10.1242/dev.01530 · Cited by 117 · see citing works
COUP-TFs regulate eye development by controlling factors essential for optic vesicle morphogenesis
Tang, K., Xie, X., Park, J., Jamrich, M., Tsai, S. Y., & Tsai, M. (2010) · Development · doi:10.1242/dev.040568 · Cited by 116 · see citing works
COUP-TF Upregulates NGFI-A Gene Expression through an Sp1 Binding Site
Pipaón, C., Tsai, S. Y., & Tsai, M. (1999) · Molecular and Cellular Biology · doi:10.1128/mcb.19.4.2734 · Cited by 110 · see citing works
Mikkola, I., Bruun, J., Holm, T., & Johansen, T. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m008882200 · Cited by 84 · see citing works
Akamatsu, W., DeVeale, B., Okano, H., Cooney, A. J., & van der Kooy, D. (2009) · The Journal of Neuroscience · doi:10.1523/jneurosci.4527-08.2009 · Cited by 72 · see citing works
MicroRNAs in early vertebrate development
Rosa, A., & Brivanlou, A. H. (2009) · Cell Cycle · doi:10.4161/cc.8.21.9847 · Cited by 63 · see citing works
Self-citationIsolation, Characterization, and Chromosomal Localization of Mouse and Human COUP-TF I and II Genes
Qiu, Y., Krishnan, V., Zeng, Z., et al. (1995) · Genomics · doi:10.1006/geno.1995.1237 · Cited by 51 · see citing works
The germ cell nuclear factor (GCNF)
Zechel, C. (2005) · Molecular Reproduction and Development · doi:10.1002/mrd.20377 · Cited by 44 · see citing works
Regulation of Oct-4 gene expression during differentiation of EC cells
Schoorlemmer, J., Jonk, L. J. C., Shen, S., van Puijenbroek, A., Feijen, A., & Kruijer, W. (1995) · Molecular Biology Reports · doi:10.1007/bf00997235 · Cited by 37 · see citing works
Guiding embryonic stem cells towards differentiation: lessons from molecular embryology
Spagnoli, F. M., & Hemmati‐Brivanlou, A. (2006) · Current Opinion in Genetics & Development · doi:10.1016/j.gde.2006.08.004 · Cited by 35 · see citing works
The varied roles of nuclear receptors during vertebrate embryonic development
Chung, A. C.-K., & Cooney, A. J. (2003) · Nuclear Receptor Signaling · doi:10.1621/nrs.01007 · Cited by 20 · see citing works
Mechanisms of epigenetic inheritance: Copying of Polycomb repressed chromatin
Francis, N. J. (2009) · Cell Cycle · doi:10.4161/cc.8.21.9876 · Cited by 18 · see citing works
Requirement for COUP-TF1 and 2 in the temporal specification of neural stem cells in CNS development
仲, 勇. (2009) · Medical Entomology and Zoology · Cited by 18 · see citing works
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- Authors
- Rosa, A., & Brivanlou, A. H.
- Affiliations
- Rockefeller University (US)
- Corresponding author
- Brivanlou, Ali H
- Venue
- The EMBO Journal
- Published
- 2010-12-10
- Volume
- 30
- Issue
- 2
- Pages
- 237-248
- ISSN
- 0261-4189 (ISSN-L)
- 1460-2075
- PMC
- PMC3025464
- OpenAlex
- W1925932947
- Cited by
- 198
- References
- 61
- Citations by year
- 2026: 1 · 2025: 4 · 2024: 13 · 2023: 5 · 2022: 7 · 2021: 9 · 2020: 11 · 2019: 11 · 2018: 11 · 2017: 14 · 2016: 11 · 2015: 13 · 2014: 20 · 2013: 28 · 2012: 25
- FWCI
- 8.85
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- Locations
- The EMBO Journal (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- IRIS Research product catalog (Sapienza University of Rome) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Pluripotent Stem Cells Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Developmental Biology and Gene Regulation
- Life Sciences › Neuroscience › Developmental Neuroscience › Neurogenesis and neuroplasticity mechanisms
- Keywords
- miR-302, Oct4, NR2F2, COUP-TFII, human embryonic stem cells, stem cell differentiation, pluripotency, microRNA regulation, alternative splicing, neural induction, neuronal gene expression, positive feedback loop, cell fate determination
- MeSH terms
- Cell Differentiation, Humans, Luciferases, Plasmids, Blotting, Northern, Blotting, Western, Alternative Splicing, DNA Primers, Gene Expression Regulation, Developmental, Reverse Transcriptase Polymerase Chain Reaction, MicroRNAs, Chromatin Immunoprecipitation, Octamer Transcription Factor-3, COUP Transcription Factor II, Gene Regulatory Networks, Embryonic Stem Cells
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