Target Deconvolution Efforts on Wnt Pathway Screen Reveal Dual Modulation of Oxidative Phosphorylation and SERCA2.
Wnt signaling is critical for development, cell proliferation and differentiation, and mutations in this pathway resulting in constitutive signaling have been implicated in various cancers. A pathway screen using a Wnt-dependent reporter identified a chemical series based on a 1,2,3-thiadiazole-5-carboxamide (TDZ) core with sub-micromolar potency. Herein we report a comprehensive mechanism-of-action deconvolution study toward identifying the efficacy target(s) and biological implication of this chemical series involving bottom-up quantitative chemoproteomics, cell biology, and biochemical methods. Through observing the effects of our probes on metabolism and performing confirmatory cellular and biochemical assays, we found that this chemical series inhibits ATP synthesis by uncoupling the mitochondrial potential. Affinity chemoproteomics experiments identified sarco(endo)plasmic reticulum Ca2+ -dependent ATPase (SERCA2) as a binding partner of the TDZ series, and subsequent validation studies suggest that the TDZ series can act as ionophores through SERCA2 toward Wnt pathway inhibition.
['Casás-Selves|Matias|M|0000-0001-8514-0632', 'Zhang|Andrew X|AX|0000-0003-0406-2404', 'Dowling|James E|JE|0000-0002-3984-4169', 'Hallén|Stefan|S|', 'Kawatkar|Aarti|A|', 'Pace|Nicholas J|NJ|', 'Denz|Christopher R|CR|', 'Pontz|Timothy|T|', 'Garahdaghi|Farzin|F|', 'Cao|Qing|Q|', 'Sabirsh|Alan|A|', 'Thakur|Kumar|K|', "O'Connell|Nichole|N|", 'Hu|Jun|J|', 'Cornella-Taracido|Iván|I|', 'Weerapana|Eranthie|E|', 'Zinda|Michael|M|', 'Goodnow|Robert A|RA|', 'Castaldi|M Paola|MP|']
D004305:Dose-Response Relationship, Drug D006801:Humans D015394:Molecular Structure D010085:Oxidative Phosphorylation D053498:Sarcoplasmic Reticulum Calcium-Transporting ATPases D013329:Structure-Activity Relationship D013830:Thiadiazoles D060449:Wnt Signaling Pathway
Wnt signaling chemoproteomics chemical biology target identification