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 Bradley, Meghan E. McGee-Lawrence, Hee Jeong Im, Marcel Karperien, Aaron Krych, Jennifer J Westendorf, A. Noelle Larson, Andre J van Wijnen

Research output: Contribution to journalArticle

3 Citations (Scopus)

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

Fingerprint

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

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; McGee-Lawrence, Meghan E.; Im, Hee Jeong; Karperien, Marcel; Krych, Aaron; 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, E, McGee-Lawrence, ME, Im, HJ, Karperien, M, Krych, A, 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 ; McGee-Lawrence, Meghan E. ; Im, Hee Jeong ; Karperien, Marcel ; Krych, Aaron ; 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.
@article{39f608a7baee4765beb065ecd93c6d81,
title = "Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development",
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.",
author = "Camilleri, {Emily T.} and Amel Dudakovic and Riester, {Scott M.} and Catalina Galeano-Garces and Paradise, {Christopher R.} and Elizabeth Bradley and McGee-Lawrence, {Meghan E.} and Im, {Hee Jeong} and Marcel Karperien and Aaron Krych and Westendorf, {Jennifer J} and {Noelle Larson}, A. and {van Wijnen}, {Andre J}",
year = "2018",
month = "1",
day = "1",
doi = "10.1074/jbc.RA118.003909",
language = "English (US)",
volume = "293",
pages = "19001--19011",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "49",

}

TY - JOUR

T1 - Loss of histone methyltransferase Ezh2 stimulates an osteogenic transcriptional program in chondrocytes but does not affect cartilage development

AU - Camilleri, Emily T.

AU - Dudakovic, Amel

AU - Riester, Scott M.

AU - Galeano-Garces, Catalina

AU - Paradise, Christopher R.

AU - Bradley, Elizabeth

AU - McGee-Lawrence, Meghan E.

AU - Im, Hee Jeong

AU - Karperien, Marcel

AU - Krych, Aaron

AU - Westendorf, Jennifer J

AU - Noelle Larson, A.

AU - van Wijnen, Andre J

PY - 2018/1/1

Y1 - 2018/1/1

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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85058182206&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85058182206&partnerID=8YFLogxK

U2 - 10.1074/jbc.RA118.003909

DO - 10.1074/jbc.RA118.003909

M3 - Article

VL - 293

SP - 19001

EP - 19011

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 49

ER -