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Megakaryocytes maintain homeostatic quiescence and promote post-injury regeneration of hematopoietic stem cells

Nature Medicine volume 20, pages 1321–1326 (2014)Cite this article

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Abstract

Multiple bone marrow stromal cell types have been identified as hematopoietic stem cell (HSC)-regulating niche cells1,2,3,4,5,6,7. However, whether HSC progeny can serve directly as HSC niche cells has not previously been shown. Here we report a dichotomous role of megakaryocytes (MKs) in both maintaining HSC quiescence during homeostasis and promoting HSC regeneration after chemotherapeutic stress. We show that MKs are physically associated with HSCs in the bone marrow of mice and that MK ablation led to activation of quiescent HSCs and increased HSC proliferation. RNA sequencing (RNA-seq) analysis revealed that transforming growth factor β1 (encoded by Tgfb1) is expressed at higher levels in MKs as compared to other stromal niche cells. MK ablation led to reduced levels of biologically active TGF-β1 protein in the bone marrow and nuclear-localized phosphorylated SMAD2/3 (pSMAD2/3) in HSCs, suggesting that MKs maintain HSC quiescence through TGF-β–SMAD signaling8,9. Indeed, TGF-β1 injection into mice in which MKs had been ablated restored HSC quiescence, and conditional deletion of Tgfb1 in MKs increased HSC activation and proliferation. These data demonstrate that TGF-β1 is a dominant signal emanating from MKs that maintains HSC quiescence. However, under conditions of chemotherapeutic challenge, MK ablation resulted in a severe defect in HSC expansion. In response to stress, fibroblast growth factor 1 (FGF1) signaling from MKs transiently dominates over TGF-β inhibitory signaling to stimulate HSC expansion10,11. Overall, these observations demonstrate that MKs serve as HSC-derived niche cells to dynamically regulate HSC function.

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Figure 1: MK ablation impairs HSC quiescence and causes HSC proliferation.
Figure 2: MKs maintain HSC quiescence through TGF-β signaling.
Figure 3: MK ablation causes an HSC regeneration defect after 5FU treatment.
Figure 4: MKs support HSC expansion under stress through FGF1 signaling.

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Acknowledgements

We are grateful to members of the Li lab for scientific discussion and manuscript critical reading, M. Hembree, K. Zapien, D. Dukes and B. Lewis for technical support and K. Tannen for manuscript editing and proofreading. We thank H. Lin (Massachusetts General Hospital), who generously provided the CAGA-luc plasmid. We thank C. Deng (US National Institutes of Health) for providing Fgfr1flox/flox mice. This work was funded by the Stowers Institute for Medical Research.

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Authors and Affiliations

  1. Stowers Institute for Medical Research, Kansas City, Missouri, USA

    Meng Zhao, John M Perry, Heather Marshall, Aparna Venkatraman, Pengxu Qian, Xi C He & Linheng Li

  2. Centre for Stem Cell Research, Christian Medical College, Vellore, India

    Aparna Venkatraman

  3. Laboratory of Blood and Vascular Biology, Rockefeller University, New York, New York, USA

    Jasimuddin Ahamed

  4. Department of Pathology and Laboratory Medicine, University of Kansas Medical Center, Kansas City, Kansas, USA

    Linheng Li

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  1. Meng Zhao
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Contributions

M.Z. performed experiments, analyzed data and wrote the paper. J.M.P. contributed to HSC culture studies and edited the manuscript. H.M. contributed to part of the mouse work. A.V., P.Q. and X.C.H. performed RNA-seq. J.A. shared reagents. L.L. directed the overall project and co-wrote the manuscript. All authors discussed the results and commented on the manuscript.

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Correspondence to Linheng Li.

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Zhao, M., Perry, J., Marshall, H. et al. Megakaryocytes maintain homeostatic quiescence and promote post-injury regeneration of hematopoietic stem cells. Nat Med 20, 1321–1326 (2014). https://doi.org/10.1038/nm.3706

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