CoChat
Sign Up
Cited by 271Pluripotent Stem Cells ResearchDevelopmental Biology and Gene RegulationReproductive Biology and Fertility

Myc Depletion Induces a Pluripotent Dormant State Mimicking Diapause

Open access
Scognamiglio, Roberta, Cabezas-Wallscheid, Nina, Thier, Marc Christian, Altamura, Sandro, Reyes, Alejandro, Prendergast, Áine M., Baumgärtner, Daniel, Carnevalli, Larissa S. +14 more·2016·Cell
DOI 10.1016/j.cell.2015.12.033PMC PMC4752822
Open this paper in CoChat — free accountOpen PDF

Abstract

Mouse embryonic stem cells (ESCs) are maintained in a naive ground state of pluripotency in the presence of MEK and GSK3 inhibitors. Here, we show that ground-state ESCs express low Myc levels. Deletion of both c-myc and N-myc (dKO) or pharmacological inhibition of Myc activity strongly decreases transcription, splicing, and protein synthesis, leading to proliferation arrest. This process is reversible and occurs without affecting pluripotency, suggesting that Myc-depleted stem cells enter a state of dormancy similar to embryonic diapause. Indeed, c-Myc is depleted in diapaused blastocysts, and the differential expression signatures of dKO ESCs and diapaused epiblasts are remarkably similar. Following Myc inhibition, pre-implantation blastocysts enter biosynthetic dormancy but can progress through their normal developmental program after transfer into pseudo-pregnant recipients. Our study shows that Myc controls the biosynthetic machinery of stem cells without affecting their potency, thus regulating their entry and exit from the dormant state.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.1016/j.cell.2015.12.033

Cites

6 of 6 references verified0 citation errors · 0 not checkable

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Previous observations correlate cell size and biosynthetic activity.

    “consistent with previous observations correlating cell size and biosynthetic activity (Fig. S3 B; Iritani and Eisenman, 1999; Scognamiglio et al., 2016)”

    From PU.1 enforces quiescence and limits hematopoietic stem cell expansion during inflammatory stress (2021) · cites this paper as Scognamiglio et al., 2016

    The Results section reports that Myc-inhibited embryonic stem cells formed small colonies and showed drastically reduced DNA and protein synthesis. Small colonies do not establish smaller individual cells, however, so the paper’s results do not directly show a correlation between cell size and biosynthetic activity. Its References section lists an earlier study whose title links protein synthesis and cell size. (LLM, conf=0.78)

    Experimental — accuracy not yet measured

  • IL-1 downregulates expression of Myc, a crucial upstream regulator of protein synthesis and cell cycle genes that leads to defective cell cycle entry if deleted in stem cells.

    “Likewise, IL-1 downregulates expression of Myc, a crucial upstream regulator of protein synthesis and cell cycle genes that leads to defective cell cycle entry if deleted in stem cells (Laurenti et al., 2008; Scognamiglio et al., 2016; Wilson et al., 2004).”

    From PU.1 enforces quiescence and limits hematopoietic stem cell expansion during inflammatory stress (2021) · cites this paper as Scognamiglio et al., 2016

    The Results sections report that loss of Myc in embryonic stem cells downregulates biosynthetic and DNA-replication genes, reduces protein synthesis, and causes cell-cycle arrest. Our copy does not show that IL-1 downregulates Myc, so it supports the consequences of Myc loss but not the sentence’s IL-1 premise. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • Oma and IL-1 reduced the forward scatter (FSC) profile of cultured HSCs.

    “Oma and IL-1 also reduced the forward scatter (FSC) profile of cultured HSCs, consistent with previous observations correlating cell size and biosynthetic activity (Fig. S3 B; Iritani and Eisenman, 1999; Scognamiglio et al., 2016).”

    From PU.1 enforces quiescence and limits hematopoietic stem cell expansion during inflammatory stress (2021) · cites this paper as Scognamiglio et al., 2016

    The citation accompanies a statement about prior observations linking cell size and biosynthetic activity; the Oma/IL-1 FSC result is presented as the citing authors’ own finding. The Results section ‘The Dormant State Induced by Myc Depletion Is Reversible’ reports smaller colonies and reduced DNA and protein synthesis in Myc-inhibited embryonic stem cells, but our copy does not report a cell-size or FSC measurement linking size to biosynthetic activity. Searches for forward scatter, cell size, and Oma/IL-1 found no such result. (LLM, conf=0.76)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

Cites

Flagged first, then by citations

Research works printed but not linked: 62

  1. Adhikary S. Eilers M. Transcriptional regulation and transformation by Myc proteins Nat. Rev. Mol. Cell Biol. 6 2005 635 645 16064138
  2. Arnold I. Watt F.M. c-Myc activation in transgenic mouse epidermis results in mobilization of stem cells and differentiation of their progeny Curr. Biol. 11 2001 558 568 11369200
  3. Baldridge M.T. King K.Y. Boles N.C. Weksberg D.C. Goodell M.A. Quiescent haematopoietic stem cells are activated by IFN-gamma in response to chronic infection Nature 465 2010 793 797 20535209 PMC2935898
  4. Batlle-Morera L. Smith A. Nichols J. Parameters influencing derivation of embryonic stem cells from murine embryos Genesis 46 2008 758 767 18837461
  5. Blackwood E.M. Eisenman R.N. Max: a helix-loop-helix zipper protein that forms a sequence-specific DNA-binding complex with Myc Science 251 1991 1211 1217 2006410
  6. Boroviak T. Loos R. Bertone P. Smith A. Nichols J. The ability of inner-cell-mass cells to self-renew as embryonic stem cells is acquired following epiblast specification Nat. Cell Biol. 16 2014 516 528 24859004 PMC4878656
  7. Boroviak T. Loos R. Lombard P. Okahara J. Behr R. Sasaki E. Nichols J. Smith A. Bertone P. Lineage-Specific Profiling Delineates the Emergence and Progression of Naive Pluripotency in Mammalian Embryogenesis Dev. Cell 35 2015 366 382 26555056 PMC4643313
  8. Carroll P.A. Diolaiti D. McFerrin L. Gu H. Djukovic D. Du J. Cheng P.F. Anderson S. Ulrich M. Hurley J.B. Deregulated Myc requires MondoA/Mlx for metabolic reprogramming and tumorigenesis Cancer Cell 27 2015 271 285 25640402 PMC4326605
  9. Cartwright P. McLean C. Sheppard A. Rivett D. Jones K. Dalton S. LIF/STAT3 controls ES cell self-renewal and pluripotency by a Myc-dependent mechanism Development 132 2005 885 896 15673569
  10. Charron J. Malynn B.A. Fisher P. Stewart V. Jeannotte L. Goff S.P. Robertson E.J. Alt F.W. Embryonic lethality in mice homozygous for a targeted disruption of the N-myc gene Genes Dev. 6 12A 1992 2248 2257 1459450
  11. Chen J.R. Cheng J.G. Shatzer T. Sewell L. Hernandez L. Stewart C.L. Leukemia inhibitory factor can substitute for nidatory estrogen and is essential to inducing a receptive uterus for implantation but is not essential for subsequent embryogenesis Endocrinology 141 2000 4365 4372 11108244
  12. Cox J. Mann M. MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification Nat. Biotechnol. 26 2008 1367 1372 19029910
  13. Dang C.V. MYC, metabolism, cell growth, and tumorigenesis Cold Spring Harb. Perspect. Med. 3 2013 3 PMC3721271 23906881
  14. Dang C.V. Web of the extended Myc network captures metabolism for tumorigenesis Cancer Cell 27 2015 160 162 25670077
  15. Dubois N.C. Adolphe C. Ehninger A. Wang R.A. Robertson E.J. Trumpp A. Placental rescue reveals a sole requirement for c-Myc in embryonic erythroblast survival and hematopoietic stem cell function Development 135 2008 2455 2465 18550708
  16. Ehninger A. Boch T. Uckelmann H. Essers M.A. Müdder K. Sleckman B.P. Trumpp A. Posttranscriptional regulation of c-Myc expression in adult murine HSCs during homeostasis and interferon-α-induced stress response Blood 123 2014 3909 3913 24795346
  17. Eilers M. Eisenman R.N. Myc’s broad reach Genes Dev. 22 2008 2755 2766 18923074 PMC2751281
  18. Eilers M. Schirm S. Bishop J.M. The MYC protein activates transcription of the alpha-prothymosin gene EMBO J. 10 1991 133 141 1989881 PMC452621
  19. Essers M.A. Offner S. Blanco-Bose W.E. Waibler Z. Kalinke U. Duchosal M.A. Trumpp A. IFNalpha activates dormant haematopoietic stem cells in vivo Nature 458 2009 904 908 19212321
  20. Flach J. Bakker S.T. Mohrin M. Conroy P.C. Pietras E.M. Reynaud D. Alvarez S. Diolaiti M.E. Ugarte F. Forsberg E.C. Replication stress is a potent driver of functional decline in ageing haematopoietic stem cells Nature 512 2014 198 202 25079315 PMC4456040
  21. Freytag S.O. Geddes T.J. Reciprocal regulation of adipogenesis by Myc and C/EBP alpha Science 256 1992 379 382 1566086
  22. Hall J. Guo G. Wray J. Eyres I. Nichols J. Grotewold L. Morfopoulou S. Humphreys P. Mansfield W. Walker R. Oct4 and LIF/Stat3 additively induce Krüppel factors to sustain embryonic stem cell self-renewal Cell Stem Cell 5 2009 597 609 19951688
  23. Hatton K.S. Mahon K. Chin L. Chiu F.C. Lee H.W. Peng D. Morgenbesser S.D. Horner J. DePinho R.A. Expression and activity of L-Myc in normal mouse development Mol. Cell. Biol. 16 1996 1794 1804 8657155 PMC231166
  24. Hondo E. Stewart C.L. Profiling gene expression in growth-arrested mouse embryos in diapause Genome Biol. 6 2005 202 15642106 PMC549055
  25. Hsieh A.L. Walton Z.E. Altman B.J. Stine Z.E. Dang C.V. MYC and metabolism on the path to cancer Semin. Cell Dev. Biol. 43 2015 11 21 26277543 PMC4818970
  26. Huang C.Y. Bredemeyer A.L. Walker L.M. Bassing C.H. Sleckman B.P. Dynamic regulation of c-Myc proto-oncogene expression during lymphocyte development revealed by a GFP-c-Myc knock-in mouse Eur. J. Immunol. 38 2008 342 349 18196519
  27. Humbert P.O. Verona R. Trimarchi J.M. Rogers C. Dandapani S. Lees J.A. E2f3 is critical for normal cellular proliferation Genes Dev. 14 2000 690 703 10733529 PMC316459
  28. Iritani B.M. Eisenman R.N. c-Myc enhances protein synthesis and cell size during B lymphocyte development Proc. Natl. Acad. Sci. USA 96 1999 13180 13185 10557294 PMC23921
  29. Johnston L.A. Prober D.A. Edgar B.A. Eisenman R.N. Gallant P. Drosophila myc regulates cellular growth during development Cell 98 1999 779 790 10499795 PMC10176494
  30. Kim J. Woo A.J. Chu J. Snow J.W. Fujiwara Y. Kim C.G. Cantor A.B. Orkin S.H. A Myc network accounts for similarities between embryonic stem and cancer cell transcription programs Cell 143 2010 313 324 20946988 PMC3018841
  31. Knoepfler P.S. Cheng P.F. Eisenman R.N. N-myc is essential during neurogenesis for the rapid expansion of progenitor cell populations and the inhibition of neuronal differentiation Genes Dev. 16 2002 2699 2712 12381668 PMC187459
  32. Laurenti E. Varnum-Finney B. Wilson A. Ferrero I. Blanco-Bose W.E. Ehninger A. Knoepfler P.S. Cheng P.F. MacDonald H.R. Eisenman R.N. Hematopoietic stem cell function and survival depend on c-Myc and N-Myc activity Cell Stem Cell 3 2008 611 624 19041778 PMC2635113
  33. Laurenti E. Wilson A. Trumpp A. Myc’s other life: stem cells and beyond Curr. Opin. Cell Biol. 21 2009 844 854 19836223
  34. Laurenti E. Frelin C. Xie S. Ferrari R. Dunant C.F. Zandi S. Neumann A. Plumb I. Doulatov S. Chen J. CDK6 levels regulate quiescence exit in human hematopoietic stem cells Cell Stem Cell 16 2015 302 313 25704240 PMC4359055
  35. Liu J. Xu Y. Stoleru D. Salic A. Imaging protein synthesis in cells and tissues with an alkyne analog of puromycin Proc. Natl. Acad. Sci. USA 109 2012 413 418 22160674 PMC3258597
  36. Mantalenakis S.J. Ketchel M.M. Frequency and extent of delayed implantation in lactating rats and mice J. Reprod. Fertil. 12 1966 391 394 5951130
  37. Marks H. Kalkan T. Menafra R. Denissov S. Jones K. Hofemeister H. Nichols J. Kranz A. Stewart A.F. Smith A. Stunnenberg H.G. The transcriptional and epigenomic foundations of ground state pluripotency Cell 149 2012 590 604 22541430 PMC3398752
  38. Martello G. Bertone P. Smith A. Identification of the missing pluripotency mediator downstream of leukaemia inhibitory factor EMBO J. 32 2013 2561 2574 23942233 PMC3791366
  39. Nichols J. Chambers I. Smith A. Derivation of germline competent embryonic stem cells with a combination of interleukin-6 and soluble interleukin-6 receptor Exp. Cell Res. 215 1994 237 239 7957676
  40. Nichols J. Chambers I. Taga T. Smith A. Physiological rationale for responsiveness of mouse embryonic stem cells to gp130 cytokines Development 128 2001 2333 2339 11493552
  41. Oskarsson T. Batlle E. Massagué J. Metastatic stem cells: sources, niches, and vital pathways Cell Stem Cell 14 2014 306 321 24607405 PMC3998185
  42. Pelengaris S. Khan M. Evan G.I. Suppression of Myc-induced apoptosis in beta cells exposes multiple oncogenic properties of Myc and triggers carcinogenic progression Cell 109 2002 321 334 12015982
  43. Rahl P.B. Lin C.Y. Seila A.C. Flynn R.A. McCuine S. Burge C.B. Sharp P.A. Young R.A. c-Myc regulates transcriptional pause release Cell 141 2010 432 445 20434984 PMC2864022
  44. Renfree M.B. Shaw G. Diapause Annu. Rev. Physiol. 62 2000 353 375 10845095
  45. Signer R.A. Magee J.A. Salic A. Morrison S.J. Haematopoietic stem cells require a highly regulated protein synthesis rate Nature 509 2014 49 54 24670665 PMC4015626
  46. Silva J. Nichols J. Theunissen T.W. Guo G. van Oosten A.L. Barrandon O. Wray J. Yamanaka S. Chambers I. Smith A. Nanog is the gateway to the pluripotent ground state Cell 138 2009 722 737 19703398 PMC3437554
  47. Smith K.N. Singh A.M. Dalton S. Myc represses primitive endoderm differentiation in pluripotent stem cells Cell Stem Cell 7 2010 343 354 20804970 PMC2954754
  48. Sosa M.S. Bragado P. Aguirre-Ghiso J.A. Mechanisms of disseminated cancer cell dormancy: an awakening field Nat. Rev. Cancer 14 2014 611 622 25118602 PMC4230700
  49. Srinivas S. Watanabe T. Lin C.S. William C.M. Tanabe Y. Jessell T.M. Costantini F. Cre reporter strains produced by targeted insertion of EYFP and ECFP into the ROSA26 locus BMC Dev. Biol. 1 2001 4 11299042 PMC31338
  50. Stewart C.L. Kaspar P. Brunet L.J. Bhatt H. Gadi I. Köntgen F. Abbondanzo S.J. Blastocyst implantation depends on maternal expression of leukaemia inhibitory factor Nature 359 1992 76 79 1522892
  51. Takashima Y. Guo G. Loos R. Nichols J. Ficz G. Krueger F. Oxley D. Santos F. Clarke J. Mansfield W. Resetting transcription factor control circuitry toward ground-state pluripotency in human Cell 158 2014 1254 1269 25215486 PMC4162745
  52. Takubo K. Nagamatsu G. Kobayashi C.I. Nakamura-Ishizu A. Kobayashi H. Ikeda E. Goda N. Rahimi Y. Johnson R.S. Soga T. Regulation of glycolysis by Pdk functions as a metabolic checkpoint for cell cycle quiescence in hematopoietic stem cells Cell Stem Cell 12 2013 49 61 23290136 PMC6592822
  53. Trumpp A. Refaeli Y. Oskarsson T. Gasser S. Murphy M. Martin G.R. Bishop J.M. c-Myc regulates mammalian body size by controlling cell number but not cell size Nature 414 2001 768 773 11742404
  54. Varlakhanova N.V. Cotterman R.F. deVries W.N. Morgan J. Donahue L.R. Murray S. Knowles B.B. Knoepfler P.S. myc maintains embryonic stem cell pluripotency and self-renewal Differentiation 80 2010 9 19 20537458 PMC2916696
  55. Wagner A.J. Meyers C. Laimins L.A. Hay N. c-Myc induces the expression and activity of ornithine decarboxylase Cell Growth Differ. 4 1993 879 883 8297793
  56. Walter D. Lier A. Geiselhart A. Thalheimer F.B. Huntscha S. Sobotta M.C. Moehrle B. Brocks D. Bayindir I. Kaschutnig P. Exit from dormancy provokes DNA-damage-induced attrition in haematopoietic stem cells Nature 520 2015 549 552 25707806
  57. Walz S. Lorenzin F. Morton J. Wiese K.E. von Eyss B. Herold S. Rycak L. Dumay-Odelot H. Karim S. Bartkuhn M. Activation and repression by oncogenic MYC shape tumour-specific gene expression profiles Nature 511 2014 483 487 25043018 PMC6879323
  58. Wang H. Dey S.K. Roadmap to embryo implantation: clues from mouse models Nat. Rev. Genet. 7 2006 185 199 16485018
  59. Wernig M. Meissner A. Cassady J.P. Jaenisch R. c-Myc is dispensable for direct reprogramming of mouse fibroblasts Cell Stem Cell 2 2008 10 12 18371415
  60. Wilson A. Murphy M.J. Oskarsson T. Kaloulis K. Bettess M.D. Oser G.M. Pasche A.C. Knabenhans C. Macdonald H.R. Trumpp A. c-Myc controls the balance between hematopoietic stem cell self-renewal and differentiation Genes Dev. 18 2004 2747 2763 15545632 PMC528895
  61. Wilson A. Laurenti E. Trumpp A. Balancing dormant and self-renewing hematopoietic stem cells Curr. Opin. Genet. Dev. 19 2009 461 468 19811902
  62. Zirath H. Frenzel A. Oliynyk G. Segerström L. Westermark U.K. Larsson K. Munksgaard Persson M. Hultenby K. Lehtiö J. Einvik C. MYC inhibition induces metabolic changes leading to accumulation of lipid droplets in tumor cells Proc. Natl. Acad. Sci. USA 110 2013 10258 10263 23733953 PMC3690852

Cited by

Sorted by citations

Authors
Scognamiglio, R., Cabezas-Wallscheid, N., Thier, M. C., Altamura, S., Reyes, A., Prendergast, Á. M., Baumgärtner, D., Carnevalli, L. S., Atzberger, A., Haas, S., von Paleske, L., Boroviak, T., Wörsdörfer, P., Essers, M. A. G., Kloz, U., Eisenman, R. N., Edenhofer, F., Bertone, P., Huber, W., van der Hoeven, F., Smith, A., & Trumpp, A.
Affiliations
  • German Cancer Research Center (DE)
  • Heidelberg University (DE)
  • European Molecular Biology Laboratory (DE)
  • Heidelberg Institute for Stem Cell Technology and Experimental Medicine (DE)
  • Wellcome/MRC Cambridge Stem Cell Institute (GB)
  • University of Würzburg (DE)
  • Fred Hutch Cancer Center (US)
  • Universität Innsbruck (AT)
Corresponding author
Trumpp, Andreas
ORCID
Venue
Cell
Publisher
Elsevier BV
Published
2016-02
Volume
164
Issue
4
Pages
668-680
ISSN
0092-8674 (ISSN-L)
DOI
10.1016/j.cell.2015.12.033
PMC
PMC4752822
OpenAlex
W2252735470
Cited by
271
References
6
Citations by year
2026: 9 · 2025: 26 · 2024: 32 · 2023: 26 · 2022: 24 · 2021: 35 · 2020: 20 · 2019: 20 · 2018: 33 · 2017: 28 · 2016: 18
FWCI
17.16
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell (publishedVersion, cc-by-nc-nd)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by-nc-nd)
  • PubMed Central (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
  • MDC Repository (Max-Delbrueck-Center for Molecular Medicine) (acceptedVersion)
  • Online Publication Service of Würzburg University (Würzburg University) (submittedVersion, cc-by-nc-nd)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
  • Apollo (University of Cambridge) (submittedVersion, cc-by-nc-nd)
  • Apollo
Funders
  • Deutsches Krebsforschungszentrum
  • Wellcome Trust
  • Deutsche Forschungsgemeinschaft
  • Medical Research Council (MC_PC_12009)
  • Biotechnology and Biological Sciences Research Council (BB/M004023/1)
  • National Cancer Institute (R01CA057138)
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
  • Health Sciences › Medicine › Public Health, Environmental and Occupational Health › Reproductive Biology and Fertility
Keywords
c-Myc, MYCN, mouse embryonic stem cells, naive pluripotency, ground state, delayed implantation, proliferation arrest, transcription, splicing, protein synthesis, epiblast, MEK inhibition, GSK3 inhibition, stem cell metabolism
MeSH terms
Animals, Blastocyst, Embryo, Mammalian, Female, Male, Mice, Inbred C57BL, Genes, myc, Proto-Oncogene Proteins c-myc, Cell Proliferation, Mice, Embryonic Stem Cells, Gene Knockout Techniques

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review