Glycosaminoglycan silencing by engineered CXCL12 variants

FEBS Lett. 2015 Sep 14;589(19 Pt B):2819-24. doi: 10.1016/j.febslet.2015.07.052. Epub 2015 Aug 20.

Abstract

We have engineered GPCR (G protein-coupled receptor) knock-out and high GAG-binding affinity into CXCL12α to inhibit CXCL12α-induced cell migration. Compared to wtCXCL12, the mutant CXCL12α (Δ8 L29K V39K) exhibited a 5.6-fold and a 2.2-fold affinity increase for heparin and heparan sulfate, respectively. From NaCl-based heparin displacement chromatography we concluded that more amino acid replacements would lead to altered GAG (glycosaminoglycan) ligand specificity. GAG silencing by this mutant was shown in a murine seeding model of human cancer cells, whereby a greatly reduced number of liver metastases was detected when the animals were treated intravenously with 1mg/kg CXCL12α (Δ8 L29K V39K) before cancer cell application.

Keywords: Cancer therapy; Carbohydrate binding protein; Chemokine; Glycosaminoglycan; Protein engineering; Tumour metastasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Chemokine CXCL12 / genetics*
  • Chemokine CXCL12 / metabolism
  • Female
  • Gene Silencing*
  • Glycosaminoglycans / deficiency*
  • Glycosaminoglycans / genetics*
  • Humans
  • Liver / pathology
  • Mice
  • Mutation*
  • Protein Engineering*
  • Receptors, G-Protein-Coupled / metabolism
  • Signal Transduction / genetics

Substances

  • Chemokine CXCL12
  • Glycosaminoglycans
  • Receptors, G-Protein-Coupled