Integrated Transcriptome and Network Analysis Reveals Spatiotemporal Dynamics of Calvarial Suturogenesis

Cell Rep. 2020 Jul 7;32(1):107871. doi: 10.1016/j.celrep.2020.107871.


Craniofacial abnormalities often involve sutures, the growth centers of the skull. To characterize the organization and processes governing their development, we profile the murine frontal suture, a model for sutural growth and fusion, at the tissue- and single-cell level on embryonic days (E)16.5 and E18.5. For the wild-type suture, bulk RNA sequencing (RNA-seq) analysis identifies mesenchyme-, osteogenic front-, and stage-enriched genes and biological processes, as well as alternative splicing events modifying the extracellular matrix. Single-cell RNA-seq analysis distinguishes multiple subpopulations, of which five define a mesenchyme-osteoblast differentiation trajectory and show variation along the anteroposterior axis. Similar analyses of in vivo mouse models of impaired frontal suturogenesis in Saethre-Chotzen and Apert syndromes, Twist1+/- and Fgfr2+/S252W, demonstrate distinct transcriptional changes involving angiogenesis and ribogenesis, respectively. Co-expression network analysis reveals gene expression modules from which we validate key driver genes regulating osteoblast differentiation. Our study provides a global approach to gain insights into suturogenesis.

Keywords: Fgfr2; Twist1; bone; craniofacial; craniosynostosis; differential gene expression; frontal suture; mesenchyme; metopic suture; single-cell RNA-seq.

Publication types

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

MeSH terms

  • Alternative Splicing / genetics
  • Animals
  • Cell Differentiation
  • Cell Line
  • Cranial Sutures / embryology*
  • Cranial Sutures / metabolism*
  • Extracellular Matrix / genetics
  • Gene Expression Regulation, Developmental
  • Gene Regulatory Networks*
  • Humans
  • Mesoderm / metabolism
  • Mice, Inbred C57BL
  • Models, Biological
  • Osteogenesis / genetics
  • RNA-Seq
  • Single-Cell Analysis
  • Time Factors
  • Transcription, Genetic
  • Transcriptome / genetics*