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TGF-β1 impairs mechanosensation of human osteoblasts via HDAC6-mediated shortening and distortion of primary cilia

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Abstract

Transforming growth factor β (TGF-β) is a critical regulator of bone density owing to its multiple effects on cell growth and differentiation. Recently, we have shown that TGF-β1 effectively blocks bone morphogenetic protein (BMP) induced maturation of osteoblasts by upregulating histone deacetylase (HDAC) activity. The current study aimed at investigating the effect of rhTGF-β1 treatment on the expression of specific HDACs and their cellular effects, e.g., microtubule structures (primary cilia) and mechanosensation. Exposure to TGF-β1 most significantly induced expression of HDAC6 both on gene and protein level. Being most abundant in the cytoplasm HDAC6 effectively deacetylates microtubule structures. Thus, TGF-β1-induced expression of HDAC6 led to deformation and shortening of primary cilia as well as to reduced numbers of ciliated cells. Primary cilia are described to sense mechanical stimuli. Thus, fluid flow was applied to the cells, which stimulated osteoblast function (AP activity and matrix mineralization). Compromised primary cilia in TGF-β1-treated cells were associated with reduced osteogenic function, despite exposure to fluid flow conditions. Chemical inhibition of HDAC6 with Tubacin restored primary cilium structure and length. These cells showed improved osteogenic function especially under fluid flow conditions. Summarizing our results, TGF-β1 impairs human osteoblast maturation partially via HDAC6-mediated distortion and/or shortening of primary cilia. This knowledge opens up new treatment options for trauma patients with chronically elevated TGF-β1-levels (e.g., diabetics), which frequently suffer from delayed fracture healing despite adequate mechanical stimulation.

Key messages

  • Exposure to TGF-β1 induces expression of HDAC6 in human osteoblasts.

  • TGF-β1 exposed human osteoblasts show less and distorted primary cilia.

  • TGF-β1 exposed human osteoblasts are less sensitive towards mechanical stimulation.

  • Mechanosensation can be recovered by HDAC6 inhibitor Tubacin in human osteoblasts.

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Acknowledgements

We would like to thank Hanna Scheffler and Jessica Bold for their excellent technical assistance. This work was partially supported by the intramural funding of the Eberhard Karls Universität Tübingen (fortüne Junior 2174-0-0).

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Authors and Affiliations

  1. Siegfried Weller Institute for trauma research at the BG Trauma Center, Eberhard Karls Universität Tübingen, Schnarrenbergstr. 95, 72076, Tübingen, Germany

    Sabrina Ehnert, Vrinda Sreekumar, Romina H. Aspera-Werz, Sahar O. Sajadian, Elke Wintermeyer, Gunther H. Sandmann, Christian Bahrs & Andreas K. Nussler

  2. IfADo-Leibniz Research Centre for Working Environment and Human Factors at the Technical University Dortmund, Ardeystraße 67, Dortmund, Germany

    Jan G. Hengstler & Patricio Godoy

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  1. Sabrina Ehnert
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  2. Vrinda Sreekumar
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  4. Sahar O. Sajadian
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  10. Andreas K. Nussler
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Correspondence to Sabrina Ehnert.

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All human studies were performed in accordance with the 1964 Declaration of Helsinki in its latest amendment. phOBs were isolated from bone tissue of patients receiving a high tibial osteotomy in accordance with the ethical standards of the University Hospital Tübingen (364/2012BO2) and the patients’ written consent.

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Ehnert, S., Sreekumar, V., Aspera-Werz, R.H. et al. TGF-β1 impairs mechanosensation of human osteoblasts via HDAC6-mediated shortening and distortion of primary cilia. J Mol Med 95, 653–663 (2017). https://doi.org/10.1007/s00109-017-1526-4

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  • DOI: https://doi.org/10.1007/s00109-017-1526-4

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