Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Letter
  • Published:

Retinoblastoma promotes definitive erythropoiesis by repressing Id2 in fetal liver macrophages

Nature volume 432, pages 1040–1045 (2004)Cite this article

Abstract

In mammals, the fetal liver is the first site of definitive erythropoiesis—the generation of mature, enucleated red cells. The functional unit for definitive erythropoiesis is the erythroblastic island, a multicellular structure composed of a central macrophage surrounded by erythroblasts at various stages of differentiation1,2. Targeted disruption of the retinoblastoma (Rb) tumour suppressor gene in the mouse leads to embryonic death caused by failure of erythroblasts to enucleate3,4,5. The erythroid defect has been attributed to loss of Rb in cells that support erythropoiesis, but the identity of these cells is unknown6. Here we show that Rb-deficient embryos carry profound abnormalities of fetal liver macrophages that prevent physical interactions with erythroblasts. In contrast, wild-type macrophages bind Rb-deficient erythroblasts and lead them to terminal differentiation and enucleation. Loss of Id2, a helix–loop–helix protein that mediates the lethality of Rb-deficient embryos7, rescues the defects of Rb-deficient fetal liver macrophages. Rb promotes differentiation of macrophages by opposing the inhibitory functions of Id2 on the transcription factor PU.1, a master regulator of macrophage differentiation. Thus, Rb has a cell autonomous function in fetal liver macrophages, and restrains Id2 in these cells in order to implement definitive erythropoiesis.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Figure 1: Abnormal development of FLMs in Rb-/- embryos and rescue by loss of Id2.
Figure 2: Deregulated Id2 causes defective macrophage-erythroblast interactions in the Rb-deficient fetal liver.
Figure 3: Rb-deficient erythroblasts but not Rb-deficient FLMs reconstitute functional erythroblastic islands.
Figure 4: Id2 and Rb regulate transcriptional activation of CSF1R by PU.1.

Similar content being viewed by others

References

  1. Sasaki, K. & Iwatsuki, H. Origin and fate of the central macrophages of erythroblastic islands in the fetal and neonatal mouse liver. Microsc. Res. Tech. 39, 398–405 (1997)

    Article  CAS  Google Scholar 

  2. Naito, M., Hasegawa, G. & Takahashi, K. Development, differentiation, and maturation of Kupffer cells. Microsc. Res. Tech. 39, 350–364 (1997)

    Article  CAS  Google Scholar 

  3. Clarke, A. R. et al. Requirement for a functional Rb-1 gene in murine development. Nature 359, 328–330 (1992)

    Article  ADS  CAS  Google Scholar 

  4. Lee, E. Y.-H. P. et al. Mice deficient for Rb are nonviable and show defects in neurogenesis and hematopoiesis. Nature 359, 288–294 (1992)

    Article  ADS  CAS  Google Scholar 

  5. Jacks, T. et al. Effects of an Rb mutation in the mouse. Nature 359, 295–300 (1992)

    Article  ADS  CAS  Google Scholar 

  6. Vooijs, M. & Berns, A. Developmental defects and tumor predisposition in Rb mutant mice. Oncogene 18, 5293–5303 (1999)

    Article  CAS  Google Scholar 

  7. Lasorella, A., Noseda, M., Beyna, M., Yokota, Y. & Iavarone, A. Id2 is a retinoblastoma protein target and mediates signalling by Myc oncoproteins. Nature 407, 592–598 (2000)

    Article  ADS  CAS  Google Scholar 

  8. Maandag, E. C. et al. Developmental rescue of an embryonic-lethal mutation in the retinoblastoma gene in chimeric mice. EMBO J. 13, 4260–4268 (1994)

    Article  CAS  Google Scholar 

  9. Williams, B. O. et al. Extensive contribution of Rb-deficient cells to adult chimeric mice with limited histopathological consequences. EMBO J. 13, 4251–4259 (1994)

    Article  CAS  Google Scholar 

  10. Pixley, F. J. & Stanley, E. R. CSF-1 regulation of the wandering macrophage: Complexity in action. Trends Cell Biol. 114, 628–638 (2004)

    Article  Google Scholar 

  11. Cecchini, M. G. et al. Role of colony stimulating factor-1 in the establishment and regulation of tissue macrophages during postnatal development of the mouse. Development 120, 1357–1372 (1994)

    CAS  PubMed  Google Scholar 

  12. Moore, M. A. & Metcalf, D. Ontogeny of the haemopoietic system: yolk sac origin of in vivo and in vitro colony forming cells in the developing mouse embryo. Br. J. Haematol. 18, 279–296 (1970)

    Article  CAS  Google Scholar 

  13. Kawane, K. et al. Requirement of DNase II for definitive erythropoiesis in the mouse fetal liver. Science 292, 1546–1549 (2001)

    Article  ADS  CAS  Google Scholar 

  14. Morioka, Y., Naito, M., Sato, T. & Takahashi, K. Immunophenotypic and ultrastructural heterogeneity of macrophage differentiation in bone marrow and fetal hematopoiesis of mouse in vitro and in vivo. J. Leukoc. Biol. 55, 642–651 (1994)

    Article  CAS  Google Scholar 

  15. Dai, X. M. et al. Targeted disruption of the mouse colony-stimulating factor 1 receptor gene results in osteopetrosis, mononuclear phagocyte deficiency, increased primitive progenitor cell frequencies, and reproductive defects. Blood 99, 111–120 (2002)

    Article  CAS  Google Scholar 

  16. Nakajima, H. et al. All-trans and 9-cis retinoic acid enhance 1,25-dihydroxyvitamin D3-induced monocytic differentiation of U937 cells. Leuk. Res. 20, 665–676 (1996)

    Article  CAS  Google Scholar 

  17. DeKoter, R. P., Walsh, J. C. & Singh, H. PU.1 regulates both cytokine-dependent proliferation and differentiation of granulocyte/macrophage progenitors. EMBO J. 17, 4456–4468 (1998)

    Article  CAS  Google Scholar 

  18. Anderson, K. L. et al. Myeloid development is selectively disrupted in PU.1 null mice. Blood 91, 3702–3710 (1998)

    CAS  PubMed  Google Scholar 

  19. Yates, P. R., Atherton, G. T., Deed, R. W., Norton, J. D. & Sharrocks, A. D. Id helix-loop-helix proteins inhibit nucleoprotein complex formation by the TCF ETS-domain transcription factors. EMBO J. 18, 968–976 (1999)

    Article  CAS  Google Scholar 

  20. Himes, S. R. et al. A highly conserved c-fms gene intronic element controls macrophage-specific and regulated expression. J. Leukoc. Biol. 70, 812–820 (2001)

    CAS  PubMed  Google Scholar 

  21. Tagoh, H. et al. Transcription factor complex formation and chromatin fine structure alterations at the murine c-fms (CSF-1 receptor) locus during maturation of myeloid precursor cells. Genes Dev. 16, 1721–1737 (2002)

    Article  CAS  Google Scholar 

  22. Rekhtman, N. et al. PU.1 and pRB interact and cooperate to repress GATA-1 and block erythroid differentiation. Mol. Cell. Biol. 23, 7460–7474 (2003)

    Article  CAS  Google Scholar 

  23. Clark, A. J., Doyle, K. M. & Humbert, P. O. Cell-intrinsic requirement for pRb in erythropoiesis. Blood 104, 1324–1326 (2004)

    Article  CAS  Google Scholar 

  24. Bergh, G., Ehinger, M., Olsson, I., Jacobsen, S. E. & Gullberg, U. Involvement of the retinoblastoma protein in monocytic and neutrophilic lineage commitment of human bone marrow progenitor cells. Blood 94, 1971–1978 (1999)

    CAS  PubMed  Google Scholar 

  25. Tsai, K. Y. et al. Mutation of E2f-1 suppresses apoptosis and inappropriate S phase entry and extends survival of Rb-deficient mouse embryos. Mol. Cell 2, 293–304 (1998)

    Article  CAS  Google Scholar 

  26. Ziebold, U., Reza, T., Caron, A. & Lees, J. A. E2F3 contributes both to the inappropriate proliferation and to the apoptosis arising in Rb mutant embryos. Genes Dev. 15, 386–391 (2001)

    Article  CAS  Google Scholar 

  27. Wu, L. et al. Extra-embryonic function of Rb is essential for embryonic development and viability. Nature 421, 942–947 (2003)

    Article  ADS  CAS  Google Scholar 

  28. Toma, J. G., El-Bizri, H., Barnabe-Heider, F., Aloyz, R. & Miller, F. D. Evidence that helix-loop-helix proteins collaborate with retinoblastoma tumor suppressor protein to regulate cortical neurogenesis. J. Neurosci. 20, 7648–7656 (2000)

    Article  CAS  Google Scholar 

  29. Iavarone, A., Garg, P., Lasorella, A., Hsu, J. & Israel, M. A. The helix-loop-helix protein Id-2 enhances cell proliferation and binds to the retinoblastoma protein. Genes Dev. 8, 1270–1284 (1994)

    Article  CAS  Google Scholar 

  30. Stanley, E. R. Murine bone marrow-derived macrophages. Methods Mol. Biol. 75, 301–304 (1997)

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank B. Tycko for discussions, Y. Yokota and T. Jacks for providing us with Id2+/- and Rb+/- mice, T. Ludwig for support in the cultures of FLMs and erythroblasts, and R. Russell for histological consultation. We thank D. Hume for CSF-1R promoter plasmids, T. Graf for PU.1 expression plasmids, E. Knudsen for the PSM-Rb plasmid and R. Wu for help with ER-MP12 and ER-MP20 immunostaining. This work was supported by grants from NIH to A.I., A.L., G.L. and E.R.S.

Author information

Authors and Affiliations

  1. Institute for Cancer Genetics, Department of Pathology, College of Physicians and Surgeons of Columbia University, New York, New York, 10032, USA

    Antonio Iavarone & Anna Lasorella

  2. Institute for Cancer Genetics, Department of Neurology, College of Physicians and Surgeons of Columbia University, New York, New York, 10032, USA

    Antonio Iavarone

  3. Institute for Cancer Genetics, Department of Pediatrics, College of Physicians and Surgeons of Columbia University, New York, New York, 10032, USA

    Anna Lasorella

  4. Human Cancer Genetics Program, Department of Molecular Virology, Immunology and Medical Genetics, The Ohio State University, Columbus, Ohio, 43210, USA

    Gustavo Leone

  5. Department of Developmental and Molecular Biology, Albert Einstein College of Medicine, Bronx, New York, 10461, USA

    Xu-Ming Dai & E. Richard Stanley

Authors
  1. Antonio Iavarone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Emerson R. King
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Xu-Ming Dai
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gustavo Leone
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. E. Richard Stanley
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Anna Lasorella
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Antonio Iavarone.

Ethics declarations

Competing interests

The authors declare that they have no competing financial interests.

Supplementary information

Supplementary Methods

Isolation, stripping, reconstitution and functional analysis of FLM-erythroblast clusters. (DOC 34 kb)

Supplementary Figure Legends

Legends to accompany the below Supplementary Information Figures. (DOC 26 kb)

Supplementary Figure 1

Expression pattern of genes changed by more than twofold in Rb-/- MEFs compared with wild-type MEFs and reversed in Id2-/-;Rb-/- MEFs. (JPG 97 kb)

Supplementary Figure 2

Expression of CSF-1R mRNA and protein is markedly reduced in Rb-null MEFs, but not in Id2;Rb double mutant cells. (JPG 22 kb)

Supplementary Figure 3

The Helix Loop Helix (HLH) domain of Id2 is necessary for binding to PU.1 (PPT 1763 kb)

Supplementary Figure 4

Id2 inhibits binding of PU.1 to an oligonucleotide from the CSF–1R–FIRE enhancer containing two PU-boxes. (PPT 699 kb)

Supplementary Figure 5

Western blot analysis of proteins expressed in NIH 3T3 for Chromatin Immunoprecipitation. (PPT 734 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Iavarone, A., King, E., Dai, XM. et al. Retinoblastoma promotes definitive erythropoiesis by repressing Id2 in fetal liver macrophages. Nature 432, 1040–1045 (2004). https://doi.org/10.1038/nature03068

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature03068

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing