Targeting β-catenin dependent Wnt signaling via peptidomimetic inhibitors in murine chondrocytes and OA cartilage

A. Held, A. Glas, L. Dietrich, M. Bollmann, K. Brandstädter, T. N. Grossmann, C. H. Lohmann, T. Pap, J. Bertrand

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Objective: The canonical Wnt signaling pathway has been shown to be involved in regulating chondrocyte hypertrophic differentiation during Osteoarthritis (OA). The aim of this study was to test the therapeutic potential of two stapled peptide canonical Wnt inhibitors – SAH-Bcl9 and StAx-35R – in preventing Wnt induced cartilage changes in OA. Methods: Primary neonatal murine chondrocytes and cartilage explants from OA patients undergoing total joint replacement for knee OA, were used for microscopy to determine matrix and cell penetrating capacity of fluorescein isothiocyanate FITC-tagged SAH-Bcl9 and StAx-35R peptides. T cell factor/lymphoid enhancer-binding factor (TCF/LEF) reporter assays were used to monitor the inhibition of Wnt3a induced β-catenin signaling by each peptide. Changes in chondrocyte phenotypic marker gene expression were analyzed by qRT PCR. Results: Both peptides localized intercellular in primary murine chondrocytes and cartilage explants. They inhibited Wnt3a induced TCF/LEF promoter activity in primary murine chondrocytes. Both inhibitors did not rescue Wnt3a altered expression of chondrocyte phenotypic genes (Sox9, Col2a1, Acan) and hypertrophy marker gene (Col10a1) at high doses (100 ng/ml). Upon application of 10 ng/ml Wnt3a, StAx-35R partially reversed the Wnt effect on Sox9 and Col2a1 gene expression. Both peptides, however, reversed the downregulation of SOX9 and aggrecan (ACAN), and decrease of COL10A1 gene expression in preserved human OA cartilage explants. Conclusion: These data indicate that blockade of canonical Wnt signaling might be a therapeutic strategy to treat early OA cases and protect further cartilage degradation by preventing chondrocyte hypertrophic differentiation.

Original languageEnglish
Pages (from-to)818-823
Number of pages6
JournalOsteoarthritis and Cartilage
Volume26
Issue number6
Early online date17 Mar 2018
DOIs
Publication statusPublished - Jun 2018

Keywords

  • Wnt/beta-catenin signaling
  • Osteoarthritis
  • Cartilage
  • Stapled peptides
  • Chondrocytes
  • Wnt3a

Cite this

Held, A. ; Glas, A. ; Dietrich, L. ; Bollmann, M. ; Brandstädter, K. ; Grossmann, T. N. ; Lohmann, C. H. ; Pap, T. ; Bertrand, J. / Targeting β-catenin dependent Wnt signaling via peptidomimetic inhibitors in murine chondrocytes and OA cartilage. In: Osteoarthritis and Cartilage. 2018 ; Vol. 26, No. 6. pp. 818-823.
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abstract = "Objective: The canonical Wnt signaling pathway has been shown to be involved in regulating chondrocyte hypertrophic differentiation during Osteoarthritis (OA). The aim of this study was to test the therapeutic potential of two stapled peptide canonical Wnt inhibitors – SAH-Bcl9 and StAx-35R – in preventing Wnt induced cartilage changes in OA. Methods: Primary neonatal murine chondrocytes and cartilage explants from OA patients undergoing total joint replacement for knee OA, were used for microscopy to determine matrix and cell penetrating capacity of fluorescein isothiocyanate FITC-tagged SAH-Bcl9 and StAx-35R peptides. T cell factor/lymphoid enhancer-binding factor (TCF/LEF) reporter assays were used to monitor the inhibition of Wnt3a induced β-catenin signaling by each peptide. Changes in chondrocyte phenotypic marker gene expression were analyzed by qRT PCR. Results: Both peptides localized intercellular in primary murine chondrocytes and cartilage explants. They inhibited Wnt3a induced TCF/LEF promoter activity in primary murine chondrocytes. Both inhibitors did not rescue Wnt3a altered expression of chondrocyte phenotypic genes (Sox9, Col2a1, Acan) and hypertrophy marker gene (Col10a1) at high doses (100 ng/ml). Upon application of 10 ng/ml Wnt3a, StAx-35R partially reversed the Wnt effect on Sox9 and Col2a1 gene expression. Both peptides, however, reversed the downregulation of SOX9 and aggrecan (ACAN), and decrease of COL10A1 gene expression in preserved human OA cartilage explants. Conclusion: These data indicate that blockade of canonical Wnt signaling might be a therapeutic strategy to treat early OA cases and protect further cartilage degradation by preventing chondrocyte hypertrophic differentiation.",
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Held, A, Glas, A, Dietrich, L, Bollmann, M, Brandstädter, K, Grossmann, TN, Lohmann, CH, Pap, T & Bertrand, J 2018, 'Targeting β-catenin dependent Wnt signaling via peptidomimetic inhibitors in murine chondrocytes and OA cartilage' Osteoarthritis and Cartilage, vol. 26, no. 6, pp. 818-823. https://doi.org/10.1016/j.joca.2018.02.908

Targeting β-catenin dependent Wnt signaling via peptidomimetic inhibitors in murine chondrocytes and OA cartilage. / Held, A.; Glas, A.; Dietrich, L.; Bollmann, M.; Brandstädter, K.; Grossmann, T. N.; Lohmann, C. H.; Pap, T.; Bertrand, J.

In: Osteoarthritis and Cartilage, Vol. 26, No. 6, 06.2018, p. 818-823.

Research output: Contribution to JournalArticleAcademicpeer-review

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AU - Held, A.

AU - Glas, A.

AU - Dietrich, L.

AU - Bollmann, M.

AU - Brandstädter, K.

AU - Grossmann, T. N.

AU - Lohmann, C. H.

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