Publications
Publications
| Publication | Summary | Drug Discovery Phase | Topic |
|---|---|---|---|
| Lu et al. BioRxiv, 2021. Proteome-wide cellular thermal shift assay reveals novel crosstalk between brassinosteroid and auxin signaling | This study demonstrates the use of proteome-wide CETSA to identify drug targets in plants,. The research uncovered PIN-FORMED1 (PIN1) as a novel substrate of GSK3, linking brassinosteroid and auxin signaling and highlighting the importance of PIN1 phosphorylation for auxin regulation in plant vascular patterning. | Lead Generation | Mechanism of Action, Target Identification |
| Wortmann et al. Journal of Medicinal Chemistry, 2021. Discovery and Characterization of the Potent and Highly Selective1,7-Naphthyridine-Based Inhibitors BAY-091 and BAY-297 of theKinase PIP4K2A | This study identifies BAY-091 and BAY-297 as potent and selective inhibitors of PIP4K2A. While CETSA confirmed target engagement, these inhibitors did not exhibit the expected antiproliferative effects in p53-deficient tumor cells. However, BAY-091 and BAY-297 serve as useful chemical probes for studying PIP4K2A signaling in cancer and other diseases. | Target Discovery | Tool Evaluation |
| Kawatkar et al. ACS Chemical Biology, 2023. Chemical Biology Approaches Confirm MCT4 as the Therapeutic Target of a Cellular Optimized Hit | This study identifies and optimizes compounds that inhibit lactic acid efflux via MCT4, a key transporter in tumor glycolysis. Using chemical biology techniques, including CETSA, the researchers confirmed selective MCT4 engagement. These findings highlight MCT4 as a potential cancer target and demonstrate the effectiveness of orthogonal approaches for studying cellular target engagement. | Lead Optimization | Target Deconvolution |
| Perrin et al. Nat Biotechnol, 202. Identifying drug targets in tissues and whole blood with thermal-shift profiling | This study introduces tissue thermal proteome profiling (tissue-TPP) to measure drug-target interactions in vivo using mass spectrometry. By mapping protein thermal stability across organs and developing blood-CETSA and blood-TPP, the research provides insights into drug action, revealing both known and novel targets of panobinostat and vemurafenib | Lead Optimization, Pre-clinical Studies | Mechanism of Action, Target Identification, Biomarker Investigations |
| Subramanian et al.ACS Chemical Biology, 2019. Deciphering the Allosteric Binding Mechanism of the Human Tropomyosin Receptor Kinase A (hTrkA) Inhibitors | This study explores small molecule inhibitors targeting an allosteric site in hTrkA using techniques like CETSA. These inhibitors bind to the kinase’s inactive state, offering selectivity and resistance to mutations, highlighting a promising alternative to traditional active-state inhibitors for disease modification and treatment. | Lead Generation, Lead Optimization | |
| Ishii et al. Scientific Reports, 2017. CETSA quantitatively verifies in vivo target engagement of novel RIPK1 inhibitors in various biospecimens | This study demonstrates CETSA for quantitatively evaluating target engagement in mouse blood and tissues using a RIPK1 lead compound. It shows how CETSA enables SAR analysis, drug occupancy estimation, and optimized tissue homogenization for monitoring target engagement in the spleen and brain, underscoring CETSA's value in preclinical and clinical drug development. | Pre-clinical Studies | Translational Studies, In Vivo Target Engagement |
| Kawatkar et al. ACS Chem Biology, 2019. CETSA® beyond Soluble Targets: a Broad Application to Multipass Transmembrane Proteins | This study showcase CETSA's potential for studying challenging multipass transmembrane targets. Using case studies of TSPO, SERCA2, and PAR2, it highlights CETSA's ability to detect stabilization patterns and introduces modified protocols with detergent extraction to improve reliability for membrane proteins. | Target Discovery | Assay Development |
| Dziekan et al. Science Translational Medicine, 2019. Identifying purine nucleoside phosphorylase as the target of quinine using cellular thermal shift assay | This study used proteome-wide CETSA to identify drug targets in Plasmodium falciparum, validating it with known inhibitors and applying it to quinine and mefloquine. The approach identified PfPNP as a common binding target highlighting CETSA as a powerful tool for understanding antimalarial drug mechanisms and resistance. | Pre-clinical Studies | Target Identification, Translational Studies |
| Martinez Molina et al. Science, 2013. Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay | The first report on how the Cellular Thermal Shift Assay (CETSA) validates drug binding to key clinical targets, monitors drug transport, activation, off-target effects, resistance in cancer cell lines, and tissue distribution. CETSA is a valuable tool for optimizing drug target engagement. | Target Discovery, Lead Generation, Pre-clinical Studies | Tool Evaluation, Target Validation, In Vivo Target Engagement |
| Miettinen et al. EMBO Journal, 2018. Thermal proteome profiling of breast cancer cells reveals proteasomal activation by CDK4/6 inhibitor palbociclib | By utilizing thermal proteome profiling, CETSA helped identify the proteasome as a novel downstream target of palbociclib. This approach was instrumental in revealing how palbociclib activates the proteasome by reducing the association of ECM29, which normally suppresses its activity. | Pre-clinical Studies | Mechanism of Action, Target Identification, Biomarker Investigations |
| Langebäck et al. Scientific Reports, 2019. CETSA-based target engagement of taxanes as biomarkers for efficacy and resistance | This study establishes tubulin-specific CETSA as a reliable method to assess taxane target engagement and resistance in cancer treatment. Tested in breast and prostate cancer models, patient-derived xenografts, and breast cancer patients, CETSA effectively measures drug efficacy and resistance. These findings highlight CETSA's potential as a prognostic tool for optimizing taxane therapies. | Pre-clinical Studies | Biomarker Investigations |
| Graham et al. Cell Chemical Biology, 2024. Discovery of YAP1/TAZ pathway inhibitors through phenotypic screening with potent anti-tumor activity via blockade of Rho-GTPase signaling | This study identifies small molecule inhibitors of oncogenic YAP1/TAZ with potent anti-tumor activity. CETSA-MS, along with TSA and biochemical assays, confirmed BAY-856’s binding to GGTase-I, revealing its role as the compound’s direct target and helping to elucidate its molecular mechanism of action. | Lead Optimization | Target Deconvolution |
| Mazur et al. ACS Chemical Biology, 2024. Molecular Targeted Engagement of DPP9 in Rat Tissue Using CETSA, SP3 Processing, and Absolute Quantitation Mass Spectrometry | This study showcases CETSA combined with mass spectrometry for quantitative target engagement of DPP9 in rat tissue. Using ex vivo and in vivo experiments, the approach enabled dose-dependent drug-target binding analysis. This scalable method enhances drug discovery by providing clearer insights into compound screening and dose projection. | Pre-clinical Studies | Translational Studies, In Vivo Target Engagement |
| Patel et al. SLAS Discovery, 2023. Quantitative target engagement of RIPK1 in human whole blood via the cellular thermal shift assay for potential pre-clinical and clinical applications | "This study presents two quantitative CETSA assays (AlphaLISA™ and MSD) for direct target engagement in human whole blood. Using RIPK1 as a proof-of-concept, the assays demonstrated high sensitivity and robustness, making them valuable for preclinical and clinical applications. | Pre-clinical Studies | Translational Studies, In Vivo Target Engagement |
| Lu et al. Proc Natl Acad Sci U S A, 2022. Proteome-wide cellular thermal shift assay reveals unexpected cross-talk between brassinosteroid and auxin signaling | This study applies proteome-wide CETSA to Arabidopsis cells to identify targets of the GSK3 inhibitor bikinin. The results show that bikinin affects proteins involved in immune response and auxin transport, including the auxin efflux carrier PIN1. This work highlights CETSA-MS as a powerful tool for studying small molecule targets and plant signaling mechanisms. | Lead Optimization | Mechanism of Action, Target Identification |
| "Chernobrovkin et al. SLAS Discvover, 2021. A Tale of Two Tails: Efficient Profiling of Protein Degraders by Specific Functional and Target Engagement Readouts" | This study highlights the use of proteome-wide CETSA to assess target engagement and protein degradation in the development of molecular glues and PROTACs. Researchers profiled IMiDs and PROTACs, revealing their interactions with CRBN and their selective degradation effects. CETSA-MS proves to be a powerful tool in the design of more effective targeted protein degraders. | Lead Optimization | Mechanism of Action, Target Identification |
| Hendricks et al. ACS Chemical Biology, 2022. Mechanistic Insights into a CDK9 Inhibitor Via Orthogonal Proteomics Methods | This study examines the selectivity and target engagement of a CDK9 tool compound using multiple chemical biology and proteomics approaches. CETSA-MS plays a key role, confirming CDK9 as the primary target with the highest affinity. The study underscores the value of quantitative MS for proteome-wide target profiling and highlights the complementary strengths of different techniques. | Lead Optimization | Target Identification, Selectivity profiling |
| Chernobrovkin et al. bioRxiv, 2020. In-depth characterization of Staurosporine induced proteome thermal stability changes | This study optimizes CETSA coupled with mass spectrometry to track protein stability changes in response to drug treatment. By leveraging multiplexed MS, researchers analyzed broader cellular responses, including protein interactions and biomarker discovery. | Lead Generation, Lead Optimization | Selectivity Profiling |
| Shaw et al. Scientific Reports, 2018. Determining direct binders of the Androgen Receptor using a high-throughput Cellular Thermal Shift Assay | This study develops the first CETSA HT assay to identify direct androgen receptor (AR) binders in a prostate cancer cell line. Unlike traditional methods, CETSA HT distinguishes direct AR binders from co-regulator targets, providing intracellular binding affinities in a label-free, disease-relevant context. The findings highlight its potential for improving AR-targeting drug discovery. | Lead Generation | Hit Confirmation |
| Shaw et al, SLAS Discovery, 2018. Positioning High-Throughput CETSA in Early Drug Discovery through Screening against B-Raf and PARP1 | This study investigates high-throughput CETSA for measuring cellular target engagement in early drug discovery. Researchers developed CETSA HT assays for B-Raf and PARP1, confirming its alignment with traditional screening methods. The findings support CETSA HT’s use in hit identification and lead optimization. | Lead Generation | Hit Confirmation |
| Rowlands et al. SLAS Discovery, 2023.High throughput screening of 0.5 million compounds against CRAF using Alpha CETSA | This study evaluates CETSA at a high-throughput scale for early drug discovery. Researchers screened nearly 0.5 million compounds against the kinase CRAF, assessing its feasibility for hit identification. The study demonstrates CETSA’s potential for large-scale screening. | Lead Generation | Primary Screening |
| Almqvist et al, Nature Communications, 2016.CETSA screening identifies known and novel thymidylate synthase inhibitors and slow intracellular activation of 5-fluorouracil | This study presents the first library screen using CETSA of human thymidylate synthase (TS). It identified known and novel TS inhibitors, including unexpected interactions with decitabine. By tracking drug uptake and metabolism, researchers linked active metabolite formation to target binding, highlighting differences in activation kinetics. | Lead Generation | Primary Screening |
| Zhao et al. ACS Chemical Biology, 2024. Integrated Proteomics Characterization of NLRP3 Inflammasome Inhibitor MCC950 in Monocytic Cell Line Confirms Direct MCC950 Engagement with Endogenous NLRP3. | This study explores MCC950’s molecular targets and mechanism as an NLRP3 inflammasome inhibitor. CETSA and photoaffinity labeling confirmed MCC950 directly engages NLRP3, disrupting inflammasome formation. These findings clarify its pharmacological effects and support its development for inflammatory disease treatment. | Target Discovery | Tool Evaluation, Target Identification |