A Combination of rhBMP-2 (Recombinant Human Bone Morphogenetic Protein-2) and MEK (MAP Kinase/ERK Kinase) Inhibitor PD0325901 Increases Bone Formation in a Murine Model of Neurofibromatosis Type I Pseudarthrosis

J Bone Joint Surg Am. 2014 Jul 16;96(14):e117. doi: 10.2106/JBJS.M.00862.

Abstract

Background: Congenital tibial dysplasia is a severe pediatric condition that classically results in a persistent pseudarthrosis. A majority of these cases are associated with neurofibromatosis type I (NF1), a genetic disorder in which inactivation of the NF1 gene leads to overactivity of the Ras-MEK-MAPK (mitogen-activated protein kinase) signaling pathway. We therefore hypothesized that pharmaceutical inhibition of MEK-MAPK may be a beneficial therapeutic strategy.

Methods: In vitro methods were used to demonstrate a role for the MEK inhibitor PD0325901 in promoting osteogenic differentiation in Nf1-/- calvarial osteoblasts. Local applications of rhBMP-2 and/or PD0325901 were then tested in a mouse model of NF1 tibial pseudarthrosis featuring localized double inactivation of the Nf1 gene in a fracture. Mice received no treatment, PD0325901 (10 mg/kg/day from two days before fracture to ten days after fracture), rhBMP-2 (10 μg), or a combination of rhBMP-2 and PD0325901.

Results: Animals treated with the delivery vehicle alone, PD0325901, rhBMP-2, or the PD0325901 + rhBMP-2 combination showed union rates of 0%, 8%, 69% (p < 0.01), or 80% (p < 0.01), respectively, at twenty-one days after fracture. Mice treated with the rhBMP-2 + PD0325901 combination displayed a callus volume sixfold greater than the vehicle controls and twofold greater than the group receiving rhBMP-2 alone. Although MEK inhibition combined with rhBMP-2 led to increases in bone formation and union, the proportion of fibrous tissue in the callus was not significantly reduced.

Conclusions: The data suggest that MEK inhibition can promote bone formation in combination with rhBMP-2 in the context of an NF1 pseudarthrosis. However, PD0325901 did not promote substantive bone anabolism in the absence of an exogenous anabolic stimulus and did not suppress fibrosis.

Clinical relevance: This study examines a signaling pathway-based approach to treating poor bone healing in a model of NF1 pseudarthrosis.

MeSH terms

  • Animals
  • Benzamides / administration & dosage*
  • Benzamides / pharmacology
  • Bone Morphogenetic Protein 2 / administration & dosage*
  • Bone Morphogenetic Protein 2 / pharmacology
  • Diphenylamine / administration & dosage
  • Diphenylamine / analogs & derivatives*
  • Diphenylamine / pharmacology
  • Disease Models, Animal
  • Drug Therapy, Combination
  • Female
  • Mice
  • Mitogen-Activated Protein Kinase Kinases / antagonists & inhibitors*
  • Neurofibromatosis 1 / complications*
  • Osteogenesis / drug effects*
  • Pseudarthrosis / drug therapy*
  • Pseudarthrosis / etiology*
  • Recombinant Proteins / administration & dosage
  • Recombinant Proteins / pharmacology
  • Transforming Growth Factor beta / administration & dosage*
  • Transforming Growth Factor beta / pharmacology

Substances

  • Benzamides
  • Bone Morphogenetic Protein 2
  • Recombinant Proteins
  • Transforming Growth Factor beta
  • recombinant human bone morphogenetic protein-2
  • mirdametinib
  • Diphenylamine
  • Mitogen-Activated Protein Kinase Kinases