ArticleFrontiers in pharmacology2026
Quantitative high-throughput screening identifies small-molecule modulators of Wnt/β-catenin signaling.
Article in Frontiers in pharmacology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The Wnt/β-catenin signaling pathway plays an important role in development and tissue homeostasis, and its dysregulation is implicated in various pathologies, including cancer, fibrosis, and neurodegeneration. However, the discovery of small-molecule modulators of this pathway remains challenging due to the pathway's inherent complexity, characterized by ligand redundancy, overlapping receptor usage, and compensatory downstream signaling. In this study, we optimized a cell-based LEF/TCF-β-lactamase reporter assay for quantitative high-throughput screening in a 1536-well format. Screening 1280 compounds from the Library of Pharmacologically Active Compounds alongside 88 compounds from the Tox21 collection identified twelve potential antagonists of Wnt/β-catenin signaling. Follow-up studies confirmed the activity of 10 compounds, demonstrating consistent activity across two independent reporter systems (β-lactamase and luciferase). Western blot analysis showed that all compounds except for cytosine-1-beta-D-arabinofuranoside and PMEG reduced accumulation of both non-phosphorylated β-catenin (active) and total β-catenin, providing orthogonal validation of pathway inhibition. The identification of known Wnt inhibitors such as emetine, tyrphostin A9, niclosamide, ouabain, and podophyllotoxin further validated the assay's robustness. Collectively, this study establishes a robust 1536-well screening platform for identifying Wnt pathway modulators and identifies topotecan, amsacrine, brefeldin A, and tyrphostin AG 879 as candidate small-molecule antagonist, thereby expanding the chemical tools for investigating Wnt/β-catenin signaling.
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