Identifying drug targets in tissues and whole blood with thermal-shift profiling

• Published in: Nature biotechnology 2020-03, Vol.38 (3), p.303-308
• Main Authors: Perrin, Jessica, Werner, Thilo, Kurzawa, Nils, Rutkowska, Anna, Childs, Dorothee D., Kalxdorf, Mathias, Poeckel, Daniel, Stonehouse, Eugenia, Strohmer, Katrin, Heller, Bianca, Thomson, Douglas W., Krause, Jana, Becher, Isabelle, Eberl, H. Christian, Vappiani, Johanna, Sevin, Daniel C., Rau, Christina E., Franken, Holger, Huber, Wolfgang, Faelth-Savitski, Maria, Savitski, Mikhail M., Bantscheff, Marcus, Bergamini, Giovanna
• Format: Article
• Language: English
• Subjects: 631/154; 631/1647; 631/1647/2067; 631/61; 631/92; Agriculture; Animals; Azepines - administration & dosage; Azepines - pharmacology; Binding; Bioinformatics; Biological assay; Biomedical and Life Sciences; Biomedical Engineering/Biotechnology; Biomedicine; Biotechnology; Blood; Blood - metabolism; Change detection; Drug targeting; Drugs; Fatty acids; Hep G2 Cells; Histone deacetylase; Humans; In vivo methods and tests; Inhibitors; Kidney - chemistry; Kidney - metabolism; Kidneys; Letter; Life Sciences; Liver; Liver - chemistry; Liver - metabolism; Lung - chemistry; Lung - metabolism; Male; Mass Spectrometry; Mass spectroscopy; Methods; Mice; Organ Specificity; Panobinostat - administration & dosage; Panobinostat - pharmacology; Phosphorylation; Physiological aspects; Protein Stability; Proteins; Proteome - chemistry; Proteome - metabolism; Proteomes; Proteomics; Rats; Scientific imaging; Small Molecule Libraries - administration & dosage; Small Molecule Libraries - pharmacology; Spleen; Spleen - chemistry; Spleen - metabolism; Testis - chemistry; Testis - metabolism; Therapeutic targets; Thermal stability; Thermodynamics; Tissues; Triazoles - administration & dosage; Triazoles - pharmacology; Vemurafenib - administration & dosage; Vemurafenib - pharmacology; Zinc finger proteins
• ISSN: 1087-0156; 1546-1696
• DOI: 10.1038/s41587-019-0388-4

Monitoring drug–target interactions with methods such as the cellular thermal-shift assay (CETSA) is well established for simple cell systems but remains challenging in vivo. Here we introduce tissue thermal proteome profiling (tissue-TPP), which measures binding of small-molecule drugs to proteins in tissue samples from drug-treated animals by detecting changes in protein thermal stability using quantitative mass spectrometry. We report organ-specific, proteome-wide thermal stability maps and derive target profiles of the non-covalent histone deacetylase inhibitor panobinostat in rat liver, lung, kidney and spleen and of the B-Raf inhibitor vemurafenib in mouse testis. In addition, we devised blood-CETSA and blood-TPP and applied it to measure target and off-target engagement of panobinostat and the BET family inhibitor JQ1 directly in whole blood. Blood-TPP analysis of panobinostat confirmed its binding to known targets and also revealed thermal stabilization of the zinc-finger transcription factor ZNF512. These methods will help to elucidate the mechanisms of drug action in vivo. The targets of small-molecule drugs are detected in tissue and blood using thermal proteome assays.