Targeting ovarian tumor cell adhesion mediated by tissue transglutaminase

May Khanna, Bhadrani Chelladurai, Aruna Gavini, Liwei Li, Minghai Shao, David Courtney, John J. Turchi, Daniela Matei*, Samy Meroueh

*Corresponding author for this work

Research output: Contribution to journalArticle

25 Scopus citations

Abstract

Tissue transglutaminase (TG2) is a transpeptidase involved in protein cross-linking through generation of e-(γ-glutamyl)lysine isopeptide bonds. It also promotes cell adhesion through interaction with fibronectin and facilitates formation of fibronectin-integrin complexes. This interaction is involved in tumor cell adhesion to the matrix and in the process of tumor dissemination. Its inhibition by small molecules may therefore be useful in blocking metastasis. To that end, we screened more than 800,000 compounds following an in silico docking approach targeting two distinct cavities in the vicinity of the fibronectin-binding site on TG2. A total of 120 compounds were acquired and tested in cell culture-based assays for inhibition of ovarian tumor cell adhesion and proliferation. Seven compounds showed more than 50% inhibition of cell adhesion at a concentration of 25 μmol/L. A follow-up fluorescence polarization study revealed that one compound in particular (ITP-79) inhibited binding of a TG2 peptide to a 42-kDa fragment of fibronectin in a dose-dependent manner. This inhibition was confirmed in cancer cells by coimmunoprecipitation. A competition assay with surface plasmon resonance showed that ITP-79 modulated binding of TG2 to fibronectin. Direct binding of compounds that inhibited adhesion to TG2 were examined with differential scanning fluorimetry, which measures the effect of the compound on the melting temperature of the target. Two compounds, including ITP-79, reduced TG2 stabilization, mimicking the effects of GTP, a known negative allosteric regulator of TG2 enzymatic function. This suggests a potential allosteric mechanism for the compound in light of its distal target site.

Original languageEnglish (US)
Pages (from-to)626-636
Number of pages11
JournalMolecular cancer therapeutics
Volume10
Issue number4
DOIs
StatePublished - Apr 2011

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

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    Khanna, M., Chelladurai, B., Gavini, A., Li, L., Shao, M., Courtney, D., Turchi, J. J., Matei, D., & Meroueh, S. (2011). Targeting ovarian tumor cell adhesion mediated by tissue transglutaminase. Molecular cancer therapeutics, 10(4), 626-636. https://doi.org/10.1158/1535-7163.MCT-10-0912