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Nat Biotechnol. 2020 Jan 20. doi: 10.1038/s41587-019-0388-4. [Epub ahead of print]

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

Author information

1
Cellzome GmbH, GlaxoSmithKline, Heidelberg, Germany.
2
Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
3
Genome Biology Unit, European Molecular Biology Laboratory, Heidelberg, Germany. mikhail.savitski@embl.de.
4
Cellzome GmbH, GlaxoSmithKline, Heidelberg, Germany. marcus.x.bantscheff@gsk.com.
5
Cellzome GmbH, GlaxoSmithKline, Heidelberg, Germany. giovanna.2.bergamini@gsk.com.

Abstract

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.

PMID:
31959954
DOI:
10.1038/s41587-019-0388-4

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