Frontal bone healing is sensitive to wnt signaling inhibition via lentiviral-encoded beta-catenin short hairpin RNA

Lei Zhang, Leslie Chang, Jiajia Xu, Carolyn Ann Meyers, Noah Yan, Erin Zou, Catherine Ding, Kang Ting, Chia Soo, Shen Pang, Aaron James

Research output: Contribution to journalArticle

Abstract

The Wnt/β-catenin signaling pathway plays an integral role in skeletal biology, spanning from embryonic skeletal patterning through bone maintenance and bone repair. Most experimental methods to antagonize Wnt signaling in vivo are either systemic or transient, including genetic approaches, use of small-molecule inhibitors, or neutralizing antibodies. We sought to develop a novel, localized model of prolonged Wnt/β-catenin signaling blockade by the application and validation of a lentivirus encoding β-catenin short hairpin RNA (shRNA). Efficacy of lentiviral-encoded β-catenin shRNA was first confirmed in vitro using bone marrow mesenchymal stromal cells, and in vivo using an intramedullary long bone injection model in NOD SCID mice. Next, the effects of β-catenin knockdown were assessed in a calvarial bone defect model, in which the frontal bone demonstrates enhanced bone healing associated with heightened Wnt/β-catenin signaling. Lentivirus encoding either β-catenin shRNA or random sequence shRNA with enhanced green fluorescent protein (control) was injected overlying the calvaria of NOD SCID mice and bone defects were created in either the frontal or parietal bones. Among mice treated with lentivirus encoding β-catenin shRNA, frontal bone defect healing was significantly reduced by all radiographic and histologic metrics. In contrast, parietal bone healing was minimally impacted by β-catenin shRNA. In aggregate, our data document the application and validation of a lentivirus encoding β-catenin shRNA model that represents an easily replicable tool for examining the importance of locoregional Wnt/β-catenin signaling in bone biology and regeneration.

Original languageEnglish (US)
Pages (from-to)1742-1752
Number of pages11
JournalTissue Engineering - Part A
Volume24
Issue number23-24
DOIs
StatePublished - Dec 1 2018

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Keywords

  • Beta-catenin shRNA
  • Bone healing
  • Bone repair
  • Calvaria
  • Calvarial bone defect
  • Intramembranous bone
  • Membranous bone
  • Osteogenesis
  • Skull
  • Wnt signaling

ASJC Scopus subject areas

  • Bioengineering
  • Biochemistry
  • Biomaterials
  • Biomedical Engineering

Cite this

Zhang, L., Chang, L., Xu, J., Meyers, C. A., Yan, N., Zou, E., Ding, C., Ting, K., Soo, C., Pang, S., & James, A. (2018). Frontal bone healing is sensitive to wnt signaling inhibition via lentiviral-encoded beta-catenin short hairpin RNA. Tissue Engineering - Part A, 24(23-24), 1742-1752. https://doi.org/10.1089/ten.tea.2017.0465