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The E3 ligase Itch is a negative regulator of the homeostasis and function of hematopoietic stem cells

Nature Immunology volume 12, pages 399–407 (2011)Cite this article

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Abstract

Although hematopoietic stem cells (HSCs) are the most thoroughly characterized type of adult stem cell, the intricate molecular machinery that regulates their self-renewal properties remains elusive. Here we showed that the E3 ubiquitin ligase Itch negatively regulated the development and function of HSCs. Itch−/− mice had HSCs with enhanced frequency, competence and long-term repopulating activity. Itch-deficient HSCs showed accelerated proliferation rates and sustained progenitor properties, as well as more signaling by the transcription factor Notch1, due to more accumulation of activated Notch1. Knockdown of Notch1 in Itch-mutant HSCs resulted in reversion of the phenotype. Thus, we identify Itch as a previously unknown negative regulator of HSC homeostasis and function.

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Figure 1: Itch deficiency results in a greater frequency of HSCs in the bone marrow.
Figure 2: Cell-intrinsic defects and greater competence of Itch-mutant HSCs.
Figure 3: Accelerated proliferation of Itch-mutant HSCs.
Figure 4: Augmented repopulation activity of Itch−/− HSCs after myeloablation.
Figure 5: Less spontaneous differentiation by Itch-deficient HSCs in vitro.
Figure 6: Itch deficiency results in more Notch1 protein and signaling in HPCs.
Figure 7: Knockdown of Notch1 in Itch-deficient HSCs results in reversion of their phenotype.

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Acknowledgements

We thank F. Manzo for help with manuscript submission; and the Yale Cell Sorter Facility for support. Supported by the Howard Hughes Medical Institute (R.A.F.) and the administrative core of the National Institutes of Health Center for Biomedical Research Excellence at Roger Williams Medical Center (P20RR018757).

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Author notes

  1. Chozhavendan Rathinam

    Present address: Present address: National Institutes of Health, Center for Biomedical Research Excellence in Stem Cell Biology, Roger Williams Medical Center, Boston University School of Medicine, Providence, Rhode Island, USA.,

Authors and Affiliations

  1. Department of Immunobiology, Yale University School of Medicine, New Haven, Connecticut, USA

    Chozhavendan Rathinam & Richard A Flavell

  2. Department of Biological Sciences, University of South Carolina, Columbia, South Carolina, USA

    Lydia E Matesic

  3. Howard Hughes Medical Institute, New Haven, Connecticut, USA

    Richard A Flavell

Authors
  1. Chozhavendan Rathinam
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  2. Lydia E Matesic
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  3. Richard A Flavell
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Contributions

C.R. conceived of, designed and did the study, analyzed and interpreted all data, and wrote the manuscript; L.E.M. provided the Itch−/− mice and corrected the manuscript; and R.A.F. provided advice and corrected the manuscript.

Corresponding author

Correspondence to Richard A Flavell.

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The authors declare no competing financial interests.

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Rathinam, C., Matesic, L. & Flavell, R. The E3 ligase Itch is a negative regulator of the homeostasis and function of hematopoietic stem cells. Nat Immunol 12, 399–407 (2011). https://doi.org/10.1038/ni.2021

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