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

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.

Design, Synthesis, and Phenotypic Profiling of Pyrano‐Furo‐Pyridone Pseudo Natural Products

“…For both compounds at 30 and 50 mm, high biosimilarities (> 83 %) were found to reference compounds that inhibit mitochondrial respiration, autophagy, glucose uptake as well as Wnt-and Hedgehog pathway signaling. [6,31] Subsequent investigation in cell-based assays excluded autophagy and Hedgehog signaling as well as glucose uptake inhibition as potential modes of action. However, we noted that these developmental and metabolism networks are involved in the regulation of reactive oxygen species (ROS) formation.…”

Design, Synthesis, and Phenotypic Profiling of Pyrano‐Furo‐Pyridone Pseudo Natural Products

“…[6,31] Subsequent investigation in cell-based assays excluded autophagy and Hedgehog signaling as well as glucose uptake inhibition as potential modes of action. [6,31] Subsequent investigation in cell-based assays excluded autophagy and Hedgehog signaling as well as glucose uptake inhibition as potential modes of action.…”

“…Forboth compounds at 30 and 50 mm,high biosimilarities (> 83 %) were found to reference compounds that inhibit mitochondrial respiration, autophagy,g lucose uptake as well as Wnt-and Hedgehog pathway signaling. [6,31] Subsequent investigation in cell-based assays excluded autophagy and Hedgehog signaling as well as glucose uptake inhibition as potential modes of action. However,w en oted that these developmental and metabolism networks are involved in the regulation of reactive oxygen species (ROS) formation.…”

From Phenotypic Hit to Chemical Probe: Chemical Biology Approaches to Elucidate Small Molecule Action in Complex Biological Systems

“…More often than not, target identification studies yield hit lists, rather than a single hit, and complementary approaches can aid in hit prioritization [129,130]. Not discussed in detail here are computational approaches and signature databases, which have also greatly developed over the years and form an excellent additional resource to find targets or elucidate MOAs [131][132][133][134][135][136][137][138][139][140].…”