Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development

Emily T. Camilleri, Amel Dudakovic, Scott M. Riester, Catalina Galeano-Garces, Christopher R. Paradise, Elizabeth W. Bradley, Meghan Elizabeth McGee Lawrence, Hee Jeong Im, Marcel Karperien, Aaron J. Krych, Jennifer J. Westendorf, A. Noelle Larson, Andre J. Van Wijnen

Research output: Contribution to journalArticle

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Abstract

Ezh2 is a histone methyltransferase that suppresses osteoblast maturation and skeletal development. We evaluated the role of Ezh2 in chondrocyte lineage differentiation and endochondral ossification. Ezh2 was genetically inactivated in the mesenchymal, osteoblastic, and chondrocytic lineages in mice using the Prrx1-Cre, Osx1-Cre, and Col2a1-Cre drivers, respectively. WT and conditional knockout mice were phenotypically assessed by gross morphology, histology, and micro-CT imaging. Ezh2-de-ficient chondrocytes in micromass culture models were evaluated using RNA-Seq, histologic evaluation, and Western blotting. Aged mice with Ezh2 deficiency were also evaluated for premature development of osteoarthritis using radiographic analysis. Ezh2 deficiency in murine chondrocytes reduced bone density at 4 weeks of age but caused no other gross developmental effects. Knockdown of Ezh2 in chondrocyte micromass cultures resulted in a global reduction in trimethylation of histone 3 lysine 27 (H3K27me3) and altered differentiation in vitro. RNA-Seq analysis revealed enrichment of an osteogenic gene expression profile in Ezh2-deficient chondrocytes. Joint development proceeded normally in the absence of Ezh2 in chondrocytes without inducing excessive hypertrophy or premature osteoarthritis in vivo. In summary, loss of Ezh2 reduced H3K27me3 levels, increased the expression of osteogenic genes in chondrocytes, and resulted in a transient post-natal bone phenotype. Remarkably, Ezh2 activity is dispensable for normal chondrocyte maturation and endochondral ossification in vivo, even though it appears to have a critical role during early stages of mesenchymal lineage commitment.

Original languageEnglish (US)
Pages (from-to)19001-19011
Number of pages11
JournalJournal of Biological Chemistry
Volume293
Issue number49
DOIs
StatePublished - Jan 1 2018

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Cartilage
Chondrocytes
Bone
RNA
Histology
Osteoblasts
Gene expression
Histones
Lysine
Genes
Imaging techniques
Osteogenesis
Osteoarthritis
histone methyltransferase
Transcriptome
Knockout Mice
Bone Density
Hypertrophy
Western Blotting
Phenotype

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Camilleri, E. T., Dudakovic, A., Riester, S. M., Galeano-Garces, C., Paradise, C. R., Bradley, E. W., ... Van Wijnen, A. J. (2018). Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development. Journal of Biological Chemistry, 293(49), 19001-19011. https://doi.org/10.1074/jbc.RA118.003909

Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development. / Camilleri, Emily T.; Dudakovic, Amel; Riester, Scott M.; Galeano-Garces, Catalina; Paradise, Christopher R.; Bradley, Elizabeth W.; McGee Lawrence, Meghan Elizabeth; Im, Hee Jeong; Karperien, Marcel; Krych, Aaron J.; Westendorf, Jennifer J.; Noelle Larson, A.; Van Wijnen, Andre J.

In: Journal of Biological Chemistry, Vol. 293, No. 49, 01.01.2018, p. 19001-19011.

Research output: Contribution to journalArticle

Camilleri, ET, Dudakovic, A, Riester, SM, Galeano-Garces, C, Paradise, CR, Bradley, EW, McGee Lawrence, ME, Im, HJ, Karperien, M, Krych, AJ, Westendorf, JJ, Noelle Larson, A & Van Wijnen, AJ 2018, 'Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development', Journal of Biological Chemistry, vol. 293, no. 49, pp. 19001-19011. https://doi.org/10.1074/jbc.RA118.003909
Camilleri, Emily T. ; Dudakovic, Amel ; Riester, Scott M. ; Galeano-Garces, Catalina ; Paradise, Christopher R. ; Bradley, Elizabeth W. ; McGee Lawrence, Meghan Elizabeth ; Im, Hee Jeong ; Karperien, Marcel ; Krych, Aaron J. ; Westendorf, Jennifer J. ; Noelle Larson, A. ; Van Wijnen, Andre J. / Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 49. pp. 19001-19011.
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abstract = "Ezh2 is a histone methyltransferase that suppresses osteoblast maturation and skeletal development. We evaluated the role of Ezh2 in chondrocyte lineage differentiation and endochondral ossification. Ezh2 was genetically inactivated in the mesenchymal, osteoblastic, and chondrocytic lineages in mice using the Prrx1-Cre, Osx1-Cre, and Col2a1-Cre drivers, respectively. WT and conditional knockout mice were phenotypically assessed by gross morphology, histology, and micro-CT imaging. Ezh2-de-ficient chondrocytes in micromass culture models were evaluated using RNA-Seq, histologic evaluation, and Western blotting. Aged mice with Ezh2 deficiency were also evaluated for premature development of osteoarthritis using radiographic analysis. Ezh2 deficiency in murine chondrocytes reduced bone density at 4 weeks of age but caused no other gross developmental effects. Knockdown of Ezh2 in chondrocyte micromass cultures resulted in a global reduction in trimethylation of histone 3 lysine 27 (H3K27me3) and altered differentiation in vitro. RNA-Seq analysis revealed enrichment of an osteogenic gene expression profile in Ezh2-deficient chondrocytes. Joint development proceeded normally in the absence of Ezh2 in chondrocytes without inducing excessive hypertrophy or premature osteoarthritis in vivo. In summary, loss of Ezh2 reduced H3K27me3 levels, increased the expression of osteogenic genes in chondrocytes, and resulted in a transient post-natal bone phenotype. Remarkably, Ezh2 activity is dispensable for normal chondrocyte maturation and endochondral ossification in vivo, even though it appears to have a critical role during early stages of mesenchymal lineage commitment.",
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AU - Riester, Scott M.

AU - Galeano-Garces, Catalina

AU - Paradise, Christopher R.

AU - Bradley, Elizabeth W.

AU - McGee Lawrence, Meghan Elizabeth

AU - Im, Hee Jeong

AU - Karperien, Marcel

AU - Krych, Aaron J.

AU - Westendorf, Jennifer J.

AU - Noelle Larson, A.

AU - Van Wijnen, Andre J.

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N2 - Ezh2 is a histone methyltransferase that suppresses osteoblast maturation and skeletal development. We evaluated the role of Ezh2 in chondrocyte lineage differentiation and endochondral ossification. Ezh2 was genetically inactivated in the mesenchymal, osteoblastic, and chondrocytic lineages in mice using the Prrx1-Cre, Osx1-Cre, and Col2a1-Cre drivers, respectively. WT and conditional knockout mice were phenotypically assessed by gross morphology, histology, and micro-CT imaging. Ezh2-de-ficient chondrocytes in micromass culture models were evaluated using RNA-Seq, histologic evaluation, and Western blotting. Aged mice with Ezh2 deficiency were also evaluated for premature development of osteoarthritis using radiographic analysis. Ezh2 deficiency in murine chondrocytes reduced bone density at 4 weeks of age but caused no other gross developmental effects. Knockdown of Ezh2 in chondrocyte micromass cultures resulted in a global reduction in trimethylation of histone 3 lysine 27 (H3K27me3) and altered differentiation in vitro. RNA-Seq analysis revealed enrichment of an osteogenic gene expression profile in Ezh2-deficient chondrocytes. Joint development proceeded normally in the absence of Ezh2 in chondrocytes without inducing excessive hypertrophy or premature osteoarthritis in vivo. In summary, loss of Ezh2 reduced H3K27me3 levels, increased the expression of osteogenic genes in chondrocytes, and resulted in a transient post-natal bone phenotype. Remarkably, Ezh2 activity is dispensable for normal chondrocyte maturation and endochondral ossification in vivo, even though it appears to have a critical role during early stages of mesenchymal lineage commitment.

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