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Cell-Based Screening of Focused Bioactive Compound Libraries: Assessing Small Molecule Modulators of the Canonical Wnt Signaling and Autophagy-Lysosome Pathways

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Irina V. Lebedeva, Jack Coleman, Bruce Taillon, Fred Liang, Dakai Liu, Diana Hulboy and Wayne F. Patton.
Enzo Life Sciences, Farmingdale, NY 11735, USA.

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ABSTRACT

Wnt signaling has been extensively investigated as an effector of diverse physiological processes including neuronal development and plasticity, bone development and formation, as well as pathological diseases, such as cancer and neurodegeneration. Recently, autophagy has been shown to down-regulate the Wnt signal transduction pathway via targeted degradation of a key signaling protein, Dishevelled. Given the known relationship of both the Wnt pathway and autophagy to oncology and neurodegenerative disease, undertanding the interplay between the two pathways is of significant biomedical importance. Consequently, an autophagy pathway library, consisting of 96 compounds with defined autophagy-inducing or -inhibitory activity, was evaluated using a battery of cell-based assays relevant to the two pathways. A high-throughput mammalian cell-based assay utilizing a transcription-based luciferase reporter of Wnt/β-catenin signaling was employed as well as fluorescence-based assays for detection of autophagy and intracellular protein aggregate accumulation. The highlighted battery of assays should provide insight into regulatory cross-talk between the canonical Wnt and autophagy-lysosome pathways.

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