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Granulocyte-derived TNFα promotes vascular and hematopoietic regeneration in the bone marrow

Nature Medicine volume 24, pages 95–102 (2018)Cite this article

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Abstract

Endothelial cells are a critical component of the bone marrow (BM) stromal network, which maintains and regulates hematopoietic cells1,2,3,4,5,6,7,8,9. Vascular regeneration precedes, and is necessary for, successful hematopoietic stem cell (HSC) transplantation, the only cure for most hematopoietic diseases2,4. Recent data suggest that mature hematopoietic cells regulate BM stromal-cell function10,11,12,13. Whether a similar cross-talk regulates the BM vasculature is not known. Here we found that donor hematopoietic cells act on sinusoidal endothelial cells and induce host blood vessel and hematopoietic regeneration after BM transplantation in mice. Adoptive transfer of BM, but not peripheral, granulocytes prevented the death of mice transplanted with limited numbers of HSCs and accelerated recovery of host vessels and hematopoietic cells. Moreover, selective granulocyte ablation in vivo impaired vascular and hematopoietic regeneration after BM transplantation. Gene expression analyses indicated that granulocytes are the main source of the cytokine TNFα, whereas its receptor TNFR1 is selectively upregulated in regenerating blood vessels. In adoptive transfer experiments, wild type, but not Tnfa−/−, granulocytes induced vascular recovery, and wild-type granulocyte transfer did not prevent death or promote vascular regeneration in Tnfr1−/−; Tnfr2−/− mice. Thus, by delivering TNFα to endothelial cells, granulocytes promote blood vessel growth and hematopoietic regeneration. Manipulation of the cross-talk between granulocytes and endothelial cells may lead to new therapeutic approaches to improve blood vessel regeneration and increase survival and hematopoietic recovery after HSC transplantation.

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Figure 1: Donor hematopoietic cells drive endogenous vascular regeneration after transplantation.
Figure 2: Granulocytes promote vascular regeneration.
Figure 3: Granulocytes are necessary for vascular regeneration.
Figure 4: Granulocytes promote vascular regeneration via TNFα.

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Acknowledgements

We thank M. May for excellent technical support. This work was supported by the Pardee Foundation (D.L.). E.B. was funded through a T32 training grant (2T32HD007505-21) from the Center of Organogenesis at the University of Michigan. We thank M. Hoenerhoff and the rest of the University of Michigan in vivo core facility for performing pathology analyses. We thank the mouse imaging laboratory and the flow cytometry core facility at the University of Michigan for help with imaging and FACS experiments. R.K. and flow cytometry and whole-mount immunofluorescence analyses were partially supported by a core grant from the NIH to the University of Michigan Cancer Center (P30-CA46592).

Author information

Authors and Affiliations

  1. Department of Cell and Developmental Biology, University of Michigan School of Medicine, Ann Arbor, Michigan, USA

    Emily Bowers, Anastasiya Slaughter & Daniel Lucas

  2. Center for Organogenesis, University of Michigan School of Medicine, Ann Arbor, Michigan, USA

    Emily Bowers & Daniel Lucas

  3. Ruth L. and David S. Gottesman Institute for Stem Cell and Regenerative Medicine Research, Albert Einstein College of Medicine, Bronx, New York, USA

    Paul S Frenette

  4. Departments of Medicine and of Cell Biology, Albert Einstein College of Medicine, Bronx, New York, USA

    Paul S Frenette

  5. Department of Biostatistics, University of Michigan School of Public Health, Ann Arbor, Michigan, USA

    Rork Kuick

  6. The John Goldman Center for Cellular Therapy, Hammersmith Hospital, Imperial College Healthcare NHS Trust, London, UK

    Oscar M Pello

  7. The University of Michigan Comprehensive Cancer Center, University of Michigan, Ann Arbor, USA

    Daniel Lucas

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Contributions

E.B. and D.L. designed the study; E.B., A.S. and D.L. designed, performed and analyzed experiments. O.M.P. suggested and designed experiments. R.K. performed statistical analyses. P.S.F. provided Nestin-gfp, Tnfa−/−, Tnfr1−/− and Tnfr2−/− mice and designed experiments. E.B. and D.L. wrote the manuscript with help from all coauthors. D.L. supervised the manuscript preparation.

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Correspondence to Daniel Lucas.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Text and Figures

Supplementary Figures 1–9 and Supplementary Table 1 (PDF 2606 kb)

Life Sciences Reporting Summary (PDF 251 kb)

3D reconstruction of endothelial cells (CD31/CD144+, white) in the sternum of a lethally irradiated mouse fourteen days after transplantation of 20×106 BMNC.

True vessels (with lumen) are indicated by blue arrows and can be easily distinguished from vascular sheets that appear after irradiation (yellow arrows). (MOV 1032 kb)

3D reconstruction of endothelial cells (CD31/CD144+, white) in the sternum of a lethally irradiated mouse fourteen days after transplantation of 0.1×106 BMNC.

True vessels (with lumen) are indicated by blue arrows and can be easily distinguished from vascular sheets that appear after irradiation (yellow arrows). (MOV 1007 kb)

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Bowers, E., Slaughter, A., Frenette, P. et al. Granulocyte-derived TNFα promotes vascular and hematopoietic regeneration in the bone marrow. Nat Med 24, 95–102 (2018). https://doi.org/10.1038/nm.4448

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