Related Experiment Video
Updated: Aug 14, 2026
Using Real-Time Cell Metabolic Flux Analyzer to Monitor Osteoblast Bioenergetics
Published on: March 1, 2022
Target Deconvolution Efforts on Wnt Pathway Screen Reveal Dual Modulation of Oxidative Phosphorylation and SERCA2
Matias Casás-Selves1,2, Andrew X Zhang3, James E Dowling1
1Oncology, Innovative Medicines and Early Discovery Unit, AstraZeneca, 35 Gatehouse Drive, Waltham, MA, 02451, USA.
Abstract:
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.
Insights
This study identifies a new chemical series that inhibits cancer-linked Wnt signaling by disrupting ATP synthesis and targeting the SERCA2 protein. These 1,2,3-thiadiazole-5-carboxamides act as ionophores, impacting cellular energy production.
Area of Science:
- Biochemistry
- Molecular Biology
- Oncology
Background:
- Wnt signaling is crucial for cellular processes and its dysregulation is linked to cancer.
- A Wnt-pathway screen identified 1,2,3-thiadiazole-5-carboxamides (TDZ) as potent inhibitors.
- The precise mechanism of action for this TDZ chemical series was previously unknown.
Purpose of the Study:
- To elucidate the mechanism of action of the TDZ chemical series.
- To identify the specific molecular targets responsible for TDZ's efficacy.
- To understand the biological implications of TDZ engagement with its targets.
Main Methods:
- Utilized bottom-up quantitative chemoproteomics to identify binding partners.
- Employed cell biology and biochemical assays to validate target engagement and functional effects.
- Investigated the impact of TDZ compounds on cellular metabolism and mitochondrial potential.
Main Results:
- The TDZ series was found to inhibit ATP synthesis by uncoupling mitochondrial membrane potential.
- Affinity chemoproteomics identified sarco(endo)plasmic reticulum Ca2+-dependent ATPase (SERCA2) as a direct binding partner.
- TDZ compounds function as ionophores through SERCA2, leading to Wnt pathway inhibition.
Conclusions:
- The TDZ chemical series inhibits Wnt signaling through a novel mechanism involving SERCA2 ionophore activity.
- This mechanism disrupts cellular energy homeostasis by inhibiting ATP production.
- The findings provide a deeper understanding of Wnt pathway regulation and potential therapeutic strategies for cancers driven by Wnt pathway activation.
More Related Videos
08:15Exploring Mitochondrial Energy Metabolism of Single 3D Microtissue Spheroids Using Extracellular Flux Analysis
Published on: February 3, 2022
14:39Cholinergic Ligand–dependent Modulation of Oxidative Phosphorylation Coupling in Digitonin-permeabilized BE(2)-C Neuroblastoma Cells
Published on: April 28, 2026
