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Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts

Nature volume 401, pages 670–677 (1999)Cite this article

Abstract

Id proteins may control cell differentiation by interfering with DNA binding of transcription factors. Here we show that targeted disruption of the dominant negative helix–loop–helix proteins Id1 and Id3 in mice results in premature withdrawal of neuroblasts from the cell cycle and expression of neural-specific differentiation markers. The Id1–Id3 double knockout mice also display vascular malformations in the forebrain and an absence of branching and sprouting of blood vessels into the neuroectoderm. As angiogenesis both in the brain and in tumours requires invasion of avascular tissue by endothelial cells, we examined the Id knockout mice for their ability to support the growth of tumour xenografts. Three different tumours failed to grow and/or metastasize in Id1+/-Id3-/- mice, and any tumour growth present showed poor vascularization and extensive necrosis. Thus, the Id genes are required to maintain the timing of neuronal differentiation in the embryo and invasiveness of the vasculature. Because the Id genes are expressed at very low levels in adults, they make attractive new targets for anti-angiogenic drug design.

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Figure 1: Id1-/-Id3-/- embryos exhibit cavitational lesions, cranial haemorrhage and premature neuronal differentiation.
Figure 2: Premature and ectopic expression of NeuroD1, MATH1, MATH2 and MATH3 in E11.5 Id1-/-Id3-/- embryonic brain.
Figure 3: Defects of angiogenesis in Id1-/-Id3-/- embryos and Id expression in blood vessels.
Figure 4: Tumour growth is inhibited in Id knockout mice.
Figure 5: Decreased vascularization of xenografts and reduced metastases in Id1+/- Id3-/- mutant mice.
Figure 6: B6RV2 xenografts in Id1+/-Id3-/- mice show reduced αv-integrin and MMP2 staining and a thickening of the extracellular matrix.

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Acknowledgements

We thank F. Giancotti, E. Lai and K. Hajjar for critical reading of the manuscript and members of the Benezra lab for helpful comments throughout the course of the work. S. Kerns, K. Witty-Blease, J. Waka and A. Nicola of the Molecular Cytology Core Facility at SKI provided invaluable technical assistance. We thank R. Alani for establishing the B-CA cell line, W. Mark for the B6RV2 cell line, L. Cohen-Gould for help with electron microscopy, H. Nguyen and H. Gultekin for histological consultation and S. Doshi for assistance with eye surgery. R.B. thanks J. Benezra for unwavering support and all participants of the Annual Harold Weintraub Memorial Symposium in Aspen Colorado for their insightful comments. This work was supported by grants from the Children's Brain Tumor Foundation and from the NIH (K12) (D.L.), a grant from the NIH Medical Scientist Training Program (A.Y.), a grant from NIH Cancer Center Support (K.M.) and grants from the NIH and NSF (R.B.).

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  1. David Lyden, Alison Z. Young and Katia Manova: These authors contributed equally to this work

Authors and Affiliations

  1. Departments of Cell Biology, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    Alison Z. Young, Wei Yan & Robert Benezra

  2. Departments of Molecular Cytology Core Facility, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    Katia Manova

  3. Departments of Pathology, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    William Gerald

  4. Departments of Pediatrics, Memorial Sloan-Kettering Cancer Center, New York, 10021, New York, USA

    David Lyden & Richard O'Reilly

  5. Department of Pathology, Division of Neuropathology, Kaplan Cancer Center, New York, 10016, New York, USA

    David Zagzag

  6. Howard Hughes Medical Institute for Cancer Research and Department of Biology, Massachusetts Institute of Technology, Cambridge, 02139, Massachusetts, USA

    Richard O. Hynes

  7. Department of Proteinchemistry, Max-Planck-Institute for Biochemistry, Martinsried near Munich, D-82152, Germany

    Bernhard L. Bader

  8. Department of Immunology, Duke University Medical Center, Durham, 27710, North Carolina, USA

    Yuan Zhuang

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Correspondence to Robert Benezra.

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Lyden, D., Young, A., Zagzag, D. et al. Id1 and Id3 are required for neurogenesis, angiogenesis and vascularization of tumour xenografts. Nature 401, 670–677 (1999). https://doi.org/10.1038/44334

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