Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential

Pavitra K. Jadaun; Shuang Zhang; Marijke Koedam; Jeroen Demmers; Suvro Chatterjee; Johannes P. van Leeuwen; Bram C. van der Eerden

doi:10.1002/jcp.30617

Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential

Pavitra K. Jadaun, Shuang Zhang, Marijke Koedam, Jeroen Demmers, Suvro Chatterjee, Johannes P. van Leeuwen, Bram C. van der Eerden^*

^*Corresponding author for this work

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Abstract

The bone microenvironment is one of the most hypoxic regions of the human body and in experimental models; hypoxia inhibits osteogenic differentiation of mesenchymal stromal cells (MSCs). Our previous work revealed that Mucin 1 (MUC1) was dynamically expressed during osteogenic differentiation of human MSCs and upregulated by hypoxia. Upon stimulation, its C-terminus (MUC1-CT) is proteolytically cleaved, translocases to the nucleus, and binds to promoters of target genes. Therefore, we assessed the MUC1-mediated effect of hypoxia on the proteomic composition of human osteoblast-derived extracellular matrices (ECMs) and characterized their osteogenic and angiogenic potentials in the produced ECMs. We generated ECMs from osteogenically differentiated human MSC cultured in vitro under 20% or 2% oxygen with or without GO-201, a MUC1-CT inhibitor. Hypoxia upregulated MUC1, vascular endothelial growth factor, and connective tissue growth factor independent of MUC1 inhibition, whereas GO-201 stabilized hypoxia-inducible factor 1-alpha. Hypoxia and/or MUC1-CT inhibition reduced osteogenic differentiation of human MSC by AMP-activated protein kinase/mTORC1/S6K pathway and dampened their matrix mineralization. Hypoxia modulated ECMs by transforming growth factor-beta/Smad and phosphorylation of NFκB and upregulated COL1A1, COL5A1, and COL5A3. The ECMs of hypoxic osteoblasts reduced MSC proliferation and accelerated their osteogenic differentiation, whereas MUC1-CT-inhibited ECMs counteracted these effects. In addition, ECMs generated under MUC1-CT inhibition reduced the angiogenic potential independent of oxygen concentration. We claim here that MUC1 is critical for hypoxia-mediated changes during osteoblastogenesis, which not only alters the proteomic landscape of the ECM but thereby also modulates its osteogenic and angiogenic potentials.

Original language	English
Pages (from-to)	1440-1454
Number of pages	15
Journal	Journal of Cellular Physiology
Volume	237
Issue number	2
Early online date	22 Oct 2021
DOIs	https://doi.org/10.1002/jcp.30617
Publication status	Published - Feb 2022

Bibliographical note

Funding Information:
The European Commission Programme “H2020‐MSCA‐RISE‐2015—Proposal No. 690850—RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J‐F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis.

Funding Information:
The European Commission Programme ?H2020-MSCA-RISE-2015?Proposal No. 690850?RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J-F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis.

Publisher Copyright:
© 2021 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.

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Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potentialFinal published version, 2.42 MBLicence: CC BY

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Jadaun, P. K., Zhang, S., Koedam, M., Demmers, J., Chatterjee, S., van Leeuwen, J. P., & van der Eerden, B. C. (2022). Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential. Journal of Cellular Physiology, 237(2), 1440-1454. Advance online publication. https://doi.org/10.1002/jcp.30617

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title = "Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential",

abstract = "The bone microenvironment is one of the most hypoxic regions of the human body and in experimental models; hypoxia inhibits osteogenic differentiation of mesenchymal stromal cells (MSCs). Our previous work revealed that Mucin 1 (MUC1) was dynamically expressed during osteogenic differentiation of human MSCs and upregulated by hypoxia. Upon stimulation, its C-terminus (MUC1-CT) is proteolytically cleaved, translocases to the nucleus, and binds to promoters of target genes. Therefore, we assessed the MUC1-mediated effect of hypoxia on the proteomic composition of human osteoblast-derived extracellular matrices (ECMs) and characterized their osteogenic and angiogenic potentials in the produced ECMs. We generated ECMs from osteogenically differentiated human MSC cultured in vitro under 20% or 2% oxygen with or without GO-201, a MUC1-CT inhibitor. Hypoxia upregulated MUC1, vascular endothelial growth factor, and connective tissue growth factor independent of MUC1 inhibition, whereas GO-201 stabilized hypoxia-inducible factor 1-alpha. Hypoxia and/or MUC1-CT inhibition reduced osteogenic differentiation of human MSC by AMP-activated protein kinase/mTORC1/S6K pathway and dampened their matrix mineralization. Hypoxia modulated ECMs by transforming growth factor-beta/Smad and phosphorylation of NFκB and upregulated COL1A1, COL5A1, and COL5A3. The ECMs of hypoxic osteoblasts reduced MSC proliferation and accelerated their osteogenic differentiation, whereas MUC1-CT-inhibited ECMs counteracted these effects. In addition, ECMs generated under MUC1-CT inhibition reduced the angiogenic potential independent of oxygen concentration. We claim here that MUC1 is critical for hypoxia-mediated changes during osteoblastogenesis, which not only alters the proteomic landscape of the ECM but thereby also modulates its osteogenic and angiogenic potentials.",

author = "Jadaun, {Pavitra K.} and Shuang Zhang and Marijke Koedam and Jeroen Demmers and Suvro Chatterjee and {van Leeuwen}, {Johannes P.} and {van der Eerden}, {Bram C.}",

note = "Funding Information: The European Commission Programme “H2020‐MSCA‐RISE‐2015—Proposal No. 690850—RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J‐F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis. Funding Information: The European Commission Programme ?H2020-MSCA-RISE-2015?Proposal No. 690850?RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J-F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis. Publisher Copyright: {\textcopyright} 2021 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.",

year = "2022",

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language = "English",

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Jadaun, PK, Zhang, S , Koedam, M , Demmers, J, Chatterjee, S, van Leeuwen, JP & van der Eerden, BC 2022, 'Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential', Journal of Cellular Physiology, vol. 237, no. 2, pp. 1440-1454. https://doi.org/10.1002/jcp.30617

Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential. / Jadaun, Pavitra K.; Zhang, Shuang ; Koedam, Marijke et al.
In: Journal of Cellular Physiology, Vol. 237, No. 2, 02.2022, p. 1440-1454.

Research output: Contribution to journal › Article › Academic › peer-review

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AU - Jadaun, Pavitra K.

AU - Zhang, Shuang

AU - Koedam, Marijke

AU - Demmers, Jeroen

AU - Chatterjee, Suvro

AU - van Leeuwen, Johannes P.

AU - van der Eerden, Bram C.

N1 - Funding Information: The European Commission Programme “H2020‐MSCA‐RISE‐2015—Proposal No. 690850—RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J‐F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis. Funding Information: The European Commission Programme ?H2020-MSCA-RISE-2015?Proposal No. 690850?RUBICON, supported this work, for which P.K.J. received a Marie Curie fellowship. We are thankful to professor J-F Dufour, who provided the support for the immunoblotting experiments. The authors are also thankful to K. Bezstarosti for the mass spectrometry analysis. Publisher Copyright: © 2021 The Authors. Journal of Cellular Physiology published by Wiley Periodicals LLC.

PY - 2022/2

Y1 - 2022/2

N2 - The bone microenvironment is one of the most hypoxic regions of the human body and in experimental models; hypoxia inhibits osteogenic differentiation of mesenchymal stromal cells (MSCs). Our previous work revealed that Mucin 1 (MUC1) was dynamically expressed during osteogenic differentiation of human MSCs and upregulated by hypoxia. Upon stimulation, its C-terminus (MUC1-CT) is proteolytically cleaved, translocases to the nucleus, and binds to promoters of target genes. Therefore, we assessed the MUC1-mediated effect of hypoxia on the proteomic composition of human osteoblast-derived extracellular matrices (ECMs) and characterized their osteogenic and angiogenic potentials in the produced ECMs. We generated ECMs from osteogenically differentiated human MSC cultured in vitro under 20% or 2% oxygen with or without GO-201, a MUC1-CT inhibitor. Hypoxia upregulated MUC1, vascular endothelial growth factor, and connective tissue growth factor independent of MUC1 inhibition, whereas GO-201 stabilized hypoxia-inducible factor 1-alpha. Hypoxia and/or MUC1-CT inhibition reduced osteogenic differentiation of human MSC by AMP-activated protein kinase/mTORC1/S6K pathway and dampened their matrix mineralization. Hypoxia modulated ECMs by transforming growth factor-beta/Smad and phosphorylation of NFκB and upregulated COL1A1, COL5A1, and COL5A3. The ECMs of hypoxic osteoblasts reduced MSC proliferation and accelerated their osteogenic differentiation, whereas MUC1-CT-inhibited ECMs counteracted these effects. In addition, ECMs generated under MUC1-CT inhibition reduced the angiogenic potential independent of oxygen concentration. We claim here that MUC1 is critical for hypoxia-mediated changes during osteoblastogenesis, which not only alters the proteomic landscape of the ECM but thereby also modulates its osteogenic and angiogenic potentials.

AB - The bone microenvironment is one of the most hypoxic regions of the human body and in experimental models; hypoxia inhibits osteogenic differentiation of mesenchymal stromal cells (MSCs). Our previous work revealed that Mucin 1 (MUC1) was dynamically expressed during osteogenic differentiation of human MSCs and upregulated by hypoxia. Upon stimulation, its C-terminus (MUC1-CT) is proteolytically cleaved, translocases to the nucleus, and binds to promoters of target genes. Therefore, we assessed the MUC1-mediated effect of hypoxia on the proteomic composition of human osteoblast-derived extracellular matrices (ECMs) and characterized their osteogenic and angiogenic potentials in the produced ECMs. We generated ECMs from osteogenically differentiated human MSC cultured in vitro under 20% or 2% oxygen with or without GO-201, a MUC1-CT inhibitor. Hypoxia upregulated MUC1, vascular endothelial growth factor, and connective tissue growth factor independent of MUC1 inhibition, whereas GO-201 stabilized hypoxia-inducible factor 1-alpha. Hypoxia and/or MUC1-CT inhibition reduced osteogenic differentiation of human MSC by AMP-activated protein kinase/mTORC1/S6K pathway and dampened their matrix mineralization. Hypoxia modulated ECMs by transforming growth factor-beta/Smad and phosphorylation of NFκB and upregulated COL1A1, COL5A1, and COL5A3. The ECMs of hypoxic osteoblasts reduced MSC proliferation and accelerated their osteogenic differentiation, whereas MUC1-CT-inhibited ECMs counteracted these effects. In addition, ECMs generated under MUC1-CT inhibition reduced the angiogenic potential independent of oxygen concentration. We claim here that MUC1 is critical for hypoxia-mediated changes during osteoblastogenesis, which not only alters the proteomic landscape of the ECM but thereby also modulates its osteogenic and angiogenic potentials.

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Inhibition of hypoxia-induced Mucin 1 alters the proteomic composition of human osteoblast-produced extracellular matrix, leading to reduced osteogenic and angiogenic potential

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