Gene expression signatures of a fibroblastoid preosteoblast and cuboidal osteoblast cell model compared to the MLO-Y4 osteocyte cell model

Bone. 2009 Jan;44(1):32-45. doi: 10.1016/j.bone.2008.08.133. Epub 2008 Oct 7.

Abstract

In the osteoblast 2T3 cell model, 326 genes significantly increase in expression as subconfluent fibroblastic 2T3 cells become confluent and cuboidal. This gene set includes BMP2/4, Dlx2/5, Runx2, Osterix and Lrp5, as well as TGFbeta regulated genes. Both activated or total nuclear Smad158 and Smad2 levels increase as they become confluent, and beta-catenin protein expression increases as 2T3 cells become confluent, reflecting a set of genes involved in early preosteoblast to osteoblast commitment, as observed in vitro and in vivo. Gene Set Enrichment Analysis (GSEA) demonstrated that this 326 dataset is very similar to several early osteoblast geneset signatures. The MLO-Y4 cell model is a well-known in vitro osteocyte model. The MLO-Y4 expression pattern was directly compared with the 2T3 osteoblast cell model. 181 genes that are highly expressed in MLO-Y4 osteocytes compared to osteoblasts were identified. Very few genes expressed in MLO-Y4 cells are found in osteocytes directly isolate from bone, suggesting that osteocyte specific gene programs most likely require the osteocytes to be embedded in the proper mineralized matrix. The MLO-Y4 dataset includes few established in vivo osteocyte markers, but does include several transcription factors such as Vitamin D receptor, Tcf7, and Irx5, whose expression was confirmed in osteocytes in vivo. Gene expression signatures in MLO-Y4 cells, as determined by functional clustering and interaction maps, suggest active prostaglandin-PKA pathways, genes involved in dendrite formation, acute/defense response pathways, TGFbeta signaling, and interferon/chemokine pathways. GSEA demonstrated that MLO-Y4 expression pattern is similar to macrophages, mesenchymal fibroblasts, and early osteoblasts.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bone Morphogenetic Proteins / metabolism
  • Bone and Bones / metabolism
  • Cell Lineage
  • Cluster Analysis
  • Databases, Genetic
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism*
  • Gene Expression Profiling*
  • Gene Expression Regulation
  • Gene Regulatory Networks
  • In Situ Hybridization
  • Mice
  • Models, Biological*
  • Oligonucleotide Array Sequence Analysis
  • Osteocytes / metabolism*
  • Phosphoproteins / metabolism
  • Reproducibility of Results
  • Signal Transduction
  • Smad Proteins / metabolism
  • Sp7 Transcription Factor
  • Transcription Factors / metabolism
  • Transforming Growth Factor beta / metabolism
  • beta Catenin / metabolism

Substances

  • Bone Morphogenetic Proteins
  • Phosphoproteins
  • Smad Proteins
  • Sp7 Transcription Factor
  • Sp7 protein, mouse
  • Transcription Factors
  • Transforming Growth Factor beta
  • beta Catenin