title: Monitoring drug target engagement in cells and tissues using the cellular thermal shift assay.
abstract: The efficacy of therapeutics is dependent on a drug binding to its cognate target. Optimization of target engagement by drugs in cells is often challenging, because drug binding cannot be monitored inside cells. We have developed a method for evaluating drug binding to target proteins in cells and tissue samples. This cellular thermal shift assay (CETSA) is based on the biophysical principle of ligand-induced thermal stabilization of target proteins. Using this assay, we validated drug binding for a set of important clinical targets and monitored processes of drug transport and activation, off-target effects and drug resistance in cancer cell lines, as well as drug distribution in tissues. CETSA is likely to become a valuable tool for the validation and optimization of drug target engagement.
authors.affiliation: Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martinez Molina
authors.forename: Rozbeh
authors.initials: R
authors.lastname: Jafari
authors.forename: Marina
authors.initials: M
authors.lastname: Ignatushchenko
authors.forename: Takahiro
authors.initials: T
authors.lastname: Seki
authors.forename: E Andreas
authors.initials: EA
authors.lastname: Larsson
authors.forename: Chen
authors.initials: C
authors.lastname: Dan
authors.forename: Lekshmy
authors.initials: L
authors.lastname: Sreekumar
authors.forename: Yihai
authors.initials: Y
authors.lastname: Cao
authors.forename: Pär
authors.initials: P
authors.lastname: Nordlund
mesh_terms: D000964:Antimetabolites, Antineoplastic / Q000378:metabolism; D045744:Cell Line, Tumor; D004305:Dose-Response Relationship, Drug; D016903:Drug Monitoring / Q000379:methods*; D005493:Folic Acid Antagonists / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007668:Kidney / Q000378:metabolism; D008024:Ligands; D008099:Liver / Q000378:metabolism; D058990:Molecular Targeted Therapy; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D011485:Protein Binding; D055550:Protein Stability; D011506:Proteins / Q000378:metabolism*; D011799:Quinazolines / Q000378:metabolism; D013876:Thiophenes / Q000378:metabolism; D014018:Tissue Distribution
chemical_list: D000964:Antimetabolites, Antineoplastic; D005493:Folic Acid Antagonists; D008024:Ligands; D004364:Pharmaceutical Preparations; D011506:Proteins; D011799:Quinazolines; D013876:Thiophenes; C068874:raltitrexed