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Targeting skeletal endothelium to ameliorate bone loss

Nature Medicine volume 24, pages 823–833 (2018)Cite this article

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Abstract

Recent studies have identified a specialized subset of CD31hiendomucinhi (CD31hiEMCNhi) vascular endothelium that positively regulates bone formation. However, it remains unclear how CD31hiEMCNhi endothelium levels are coupled to anabolic bone formation. Mice with an osteoblast-specific deletion of Shn3, which have markedly elevated bone formation, demonstrated an increase in CD31hiEMCNhi endothelium. Transcriptomic analysis identified SLIT3 as an osteoblast-derived, SHN3-regulated proangiogenic factor. Genetic deletion of Slit3 reduced skeletal CD31hiEMCNhi endothelium, resulted in low bone mass because of impaired bone formation and partially reversed the high bone mass phenotype of Shn3−/− mice. This coupling between osteoblasts and CD31hiEMCNhi endothelium is essential for bone healing, as shown by defective fracture repair in SLIT3-mutant mice and enhanced fracture repair in SHN3-mutant mice. Finally, administration of recombinant SLIT3 both enhanced bone fracture healing and counteracted bone loss in a mouse model of postmenopausal osteoporosis. Thus, drugs that target the SLIT3 pathway may represent a new approach for vascular-targeted osteoanabolic therapy to treat bone loss.

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Fig. 1: Shn3−/− mice have higher levels of CD31hiEMCNhi endothelium.
Fig. 2: Ablation of Shn3 in osteoblasts enhances osteogenesis and angiogenesis in vivo.
Fig. 3: Inhibition of Shn3 enhances Slit3 expression in osteoblasts.
Fig. 4: Slit3−/− mice have reduced skeletal vasculature and bone mass in vivo.
Fig. 5: Osteoblast-derived Slit3 controls osteogenesis and CD31hiEMCNhi endothelium via ROBO1.
Fig. 6: Administration of recombinant SLIT3 has therapeutic effects on bone fracture healing and OVX-induced bone loss.

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  • 29 May 2018

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Acknowledgements

We thank the many individuals who provided valuable reagents. M.B.G. holds a Career Award for Medical Scientists from the Burroughs Wellcome Foundation and is supported by the Office of the Director of the NIH under award DP5OD021351, a Junior Investigator Award from the Musculoskeletal Transplant Foundation and a March of Dimes Basil O’Connor Award. J.-H.S. is supported by National Institute of Arthritis and Musculoskeletal and Skin Diseases/NIH under R01AR068983 and a pilot project program award from UMass Center for Clinical and Translational Science. A.D.L. is supported by NIH/National Heart, Lung, and Blood Institute under R01 HL126913. This content is solely the responsibility of the authors and does not necessarily represent the official views of the NIH. We thank D. Ballon, B. He, B. -S. Ding and J. McCormick and Citigroup Biomedical Imaging Core, Weill Cornell Microscopy and Image Analysis and Flow Cytometry Core Facilities for technical support.

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Authors and Affiliations

  1. Department of Pathology and Laboratory Medicine, Cornell University, New York, NY, USA

    Ren Xu, Alisha Yallowitz, Dong Yeon Shin, Shawon Debnath, Sarfaraz Lalani, Na Li, Yifang Liu, Yi Zhang, Annarita Di Lorenzo & Matthew B. Greenblatt

  2. Department of Orthopaedics, Shanghai Key Laboratory of Orthopaedic Implant, Shanghai Ninth People’s Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China

    An Qin

  3. Laboratory of Brain Development and Repair, The Rockefeller University, New York, NY, USA

    Zhuhao Wu

  4. Division of Rheumatology, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA

    Jung-Min Kim & Jae-Hyuck Shim

  5. Research Division, Hospital for Special Surgery, New York, NY, USA

    Gang Ji, Mathias P. Bostrom & Xu Yang

  6. Department of Joint Surgery, The Third Hospital of Hebei Medical University, Shijiazhuang, China

    Gang Ji

  7. Division of Adult Reconstruction and Joint Replacement, Department of Orthopaedic Surgery, Hospital for Special Surgery, New York, NY, USA

    Mathias P. Bostrom

  8. Institute for Computational Biomedicine, Cornell University, New York, NY, USA

    Chao Zhang

  9. Department of Cancer Immunology and Virology, Dana-Farber Cancer Institute and Harvard University Medical School, Boston, MA, USA

    Han Dong & Laurie H. Glimcher

  10. Department of Medicine, Harvard Medical School and Brigham and Women’s Hospital, Boston, MA, USA

    Han Dong & Laurie H. Glimcher

  11. Division of Regenerative Medicine, Department of Medicine, Ansary Stem Cell Institute, Cornell University, New York, NY, USA

    Pouneh Kermani, Michael G. Poulos & Jason M. Butler

  12. Department of Biomechanics, Hospital for Special Surgery, New York, NY, USA

    Amanda Wach

  13. Arthritis and Tissue Degeneration Program and David Z. Rosensweig Genomics Research Center, Hospital for Special Surgery, New York, NY, USA

    Kazuki Inoue & Baohong Zhao

  14. Department of Medicine, Weill Cornell Medical College, Cornell University, New York, NY, USA

    Kazuki Inoue & Baohong Zhao

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  1. Ren Xu
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Contributions

R.X. and M.B.G. designed the experimental plan. R.X. executed most experiments. A.Y., Z.W., X.Y., P.K., N.L., Y.L., A.W., Y.Z., A.D.L., J.-H.S., J.M.B. and K.I. assisted with mouse studies. D.Y.S., J.-M.K., S.L. and B.Z. conducted in vitro experiments. S.D., G.J., H.D. and M.G.P. assisted with flow cytometry analysis and cell sorting. A.Q. collected human bone fracture samples. C.Z. performed RNA-seq analysis. M.P.B., B.Z. and J.-H.S. assisted with osteoclast studies. R.X. and M.B.G wrote the manuscript. M.B.G and L.H.G. supervised the project.

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Correspondence to Laurie H. Glimcher or Matthew B. Greenblatt.

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L.H.G. is on the board of directors of and holds equity in the GlaxoSmithKline and Waters Corporations. She is also a founder of Quentis Pharmaceuticals.

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Xu, R., Yallowitz, A., Qin, A. et al. Targeting skeletal endothelium to ameliorate bone loss. Nat Med 24, 823–833 (2018). https://doi.org/10.1038/s41591-018-0020-z

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