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‐ca...

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Bibliographic Details
Published in:ChemMedChem 2017-06, Vol.12 (12), p.917-924
Main Authors: Casás‐Selves, Matias, Zhang, Andrew X., Dowling, James E., Hallén, Stefan, Kawatkar, Aarti, Pace, Nicholas J., Denz, Christopher R., Pontz, Timothy, Garahdaghi, Farzin, Cao, Qing, Sabirsh, Alan, Thakur, Kumar, O'Connell, Nichole, Hu, Jun, Cornella‐Taracido, Iván, Weerapana, Eranthie, Zinda, Michael, Goodnow, Robert A., Castaldi, M. Paola
Format: Article
Language:English
Subjects:
Adenosine triphosphatase
Affinity
Assaying
ATP
Binding
Biochemistry
Biological effects
Ca2+-transporting ATPase
Calcium
Cell proliferation
chemical biology
Chemical synthesis
chemoproteomics
Deconvolution
Differentiation
Dose-Response Relationship, Drug
Effectiveness
Humans
Inhibition
Ionophores
Metabolism
Mitochondria
Modulation
Molecular Structure
Mutation
Oxidative phosphorylation
Oxidative Phosphorylation - drug effects
Phosphorylation
Probes
Sarcoplasmic Reticulum Calcium-Transporting ATPases - metabolism
Signal transduction
Structure-Activity Relationship
target identification
Thiadiazoles - chemical synthesis
Thiadiazoles - chemistry
Thiadiazoles - pharmacology
Wnt protein
Wnt signaling
Wnt Signaling Pathway - drug effects
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Summary: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. Waste not, Wnt not: We identified a 1,2,3‐thiadiazole‐5‐carboxamide chemical series in a Wnt pathway inhibition screen. We generated chemical probes to identify the targets of this series based on our efforts to optimize potency and physical properties. Using a combination of biochemical, cell biology, and chemoproteomics methods, we found two previously unreported mechanisms for this pharmacophore: uncouplers of the mitochondrial potential and Ca2+ ionophores through interaction with SERCA2.
ISSN:1860-7179
1860-7187
DOI:10.1002/cmdc.201700028