Combined Proteomic and In Silico Target Identification Reveal a Role for 5-Lipoxygenase in Developmental Signaling Pathways

Cell Chem Biol. 2018 Sep 20;25(9):1095-1106.e23. doi: 10.1016/j.chembiol.2018.05.016. Epub 2018 Jun 28.

Abstract

Identification and validation of the targets of bioactive small molecules identified in cell-based screening is challenging and often meets with failure, calling for the development of new methodology. We demonstrate that a combination of chemical proteomics with in silico target prediction employing the SPiDER method may provide efficient guidance for target candidate selection and prioritization for experimental in-depth evaluation. We identify 5-lipoxygenase (5-LO) as the target of the Wnt pathway inhibitor Lipoxygenin. Lipoxygenin is a non-redox 5-LO inhibitor, modulates the β-catenin-5-LO complex and induces reduction of both β-catenin and 5-LO levels in the nucleus. Lipoxygenin and the structurally unrelated 5-LO inhibitor CJ-13,610 promote cardiac differentiation of human induced pluripotent stem cells and inhibit Hedgehog, TGF-β, BMP, and Activin A signaling, suggesting an unexpected and yet unknown role of 5-LO in these developmental pathways.

Keywords: Wnt pathway; cell-based screening; lipoxygenase; target identification; target prediction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Arachidonate 5-Lipoxygenase / metabolism*
  • Cell Differentiation / drug effects
  • Cell Line
  • Cell Line, Tumor
  • Computer Simulation
  • Computer-Aided Design
  • HEK293 Cells
  • Hedgehog Proteins / metabolism
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / drug effects
  • Induced Pluripotent Stem Cells / metabolism
  • Lipoxygenase Inhibitors / chemistry*
  • Lipoxygenase Inhibitors / pharmacology*
  • Mice
  • NIH 3T3 Cells
  • Proteomics / methods*
  • Signal Transduction / drug effects*
  • Wnt Signaling Pathway / drug effects

Substances

  • Hedgehog Proteins
  • Lipoxygenase Inhibitors
  • Arachidonate 5-Lipoxygenase