Enhancer of Zeste Homolog 2 Inhibition Stimulates Bone Formation and Mitigates Bone Loss Caused by Ovariectomy in Skeletally Mature Mice

J Biol Chem. 2016 Nov 18;291(47):24594-24606. doi: 10.1074/jbc.M116.740571. Epub 2016 Oct 10.

Abstract

Perturbations in skeletal development and bone degeneration may result in reduced bone mass and quality, leading to greater fracture risk. Bone loss is mitigated by bone protective therapies, but there is a clinical need for new bone-anabolic agents. Previous work has demonstrated that Ezh2 (enhancer of zeste homolog 2), a histone 3 lysine 27 (H3K27) methyltransferase, suppressed differentiation of osteogenic progenitors. Here, we investigated whether inhibition of Ezh2 can be leveraged for bone stimulatory applications. Pharmacologic inhibition and siRNA knockdown of Ezh2 enhanced osteogenic commitment of MC3T3 preosteoblasts. Next generation RNA sequencing of mRNAs and real time quantitative PCR profiling established that Ezh2 inactivation promotes expression of bone-related gene regulators and extracellular matrix proteins. Mechanistically, enhanced gene expression was linked to decreased H3K27 trimethylation (H3K27me3) near transcriptional start sites in genome-wide sequencing of chromatin immunoprecipitations assays. Administration of an Ezh2 inhibitor modestly increases bone density parameters of adult mice. Furthermore, Ezh2 inhibition also alleviated bone loss in an estrogen-deficient mammalian model for osteoporosis. Ezh2 inhibition enhanced expression of Wnt10b and Pth1r and increased the BMP-dependent phosphorylation of Smad1/5. Thus, these data suggest that inhibition of Ezh2 promotes paracrine signaling in osteoblasts and has bone-anabolic and osteoprotective potential in adults.

Keywords: epigenetics; histone; histone methylation; methyltransferase; osteoblast; osteoporosis; skeleton.

MeSH terms

  • Animals
  • Cell Line
  • Enhancer of Zeste Homolog 2 Protein / genetics
  • Enhancer of Zeste Homolog 2 Protein / metabolism*
  • Female
  • Methylation / drug effects
  • Mice
  • Osteoblasts / metabolism*
  • Osteoblasts / pathology
  • Osteogenesis*
  • Osteoporosis / metabolism*
  • Osteoporosis / pathology
  • Ovariectomy
  • Paracrine Communication*
  • RNA, Small Interfering / pharmacology
  • Receptor, Parathyroid Hormone, Type 1
  • Smad1 Protein / genetics
  • Smad1 Protein / metabolism
  • Smad5 Protein / genetics
  • Smad5 Protein / metabolism
  • Wnt Proteins / genetics
  • Wnt Proteins / metabolism

Substances

  • RNA, Small Interfering
  • Receptor, Parathyroid Hormone, Type 1
  • Smad1 Protein
  • Smad1 protein, mouse
  • Smad5 Protein
  • Smad5 protein, mouse
  • Wnt Proteins
  • Wnt10b protein, mouse
  • Enhancer of Zeste Homolog 2 Protein
  • Ezh2 protein, mouse