Identifying drug targets in tissues and whole blood with thermal-shift profiling
| Autor: | Nils Kurzawa, Marcus Bantscheff, Mikhail M. Savitski, Eugenia Stonehouse, Maria Faelth-Savitski, Holger Franken, Bianca Heller, Christina Rau, Jessica Perrin, Anna Rutkowska, H. Christian Eberl, Johanna Vappiani, Daniel C. Sévin, Isabelle Becher, Katrin Strohmer, Jana Krause, Thilo Werner, Douglas W. Thomson, Wolfgang Huber, Daniel Poeckel, Giovanna Bergamini, Mathias Kalxdorf, Dorothee Childs |
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| Rok vydání: | 2020 |
| Předmět: |
Male
Proteome medicine.drug_class Biomedical Engineering Bioengineering Drug action Kidney Applied Microbiology and Biotechnology Mass Spectrometry Small Molecule Libraries Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine In vivo Panobinostat Testis medicine Animals Humans Vemurafenib Lung Transcription factor 030304 developmental biology Whole blood 0303 health sciences Protein Stability Chemistry Histone deacetylase inhibitor Kidney metabolism Azepines Hep G2 Cells Triazoles Rats Cell biology Blood Liver Organ Specificity Thermodynamics Molecular Medicine Spleen 030217 neurology & neurosurgery Biotechnology medicine.drug |
| Zdroj: | Nature Biotechnology. 38:303-308 |
| ISSN: | 1546-1696 1087-0156 |
| DOI: | 10.1038/s41587-019-0388-4 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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