Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay

被引：1581

作者：

Molina, Daniel Martinez ^{[1
]}

Jafari, Rozbeh ^{[1
]}

Ignatushchenko, Marina ^{[1
]}

Seki, Takahiro ^{[2
]}

Larsson, E. Andreas ^{[3
]}

Dan, Chen ^{[3
]}

Sreekumar, Lekshmy ^{[3
]}

Cao, Yihai ^{[2
,4
]}

Nordlund, Par ^{[1
,3
]}

机构：

[1] Karolinska Inst, Dept Med Biochem & Biophys, SE-17177 Stockholm, Sweden

[2] Karolinska Inst, Dept Microbiol Tumor & Cell Biol, SE-17177 Stockholm, Sweden

[3] Nanyang Technol Univ, Sch Biol Sci, Singapore 639798, Singapore

[4] Linkoping Univ, Dept Med & Hlth Sci, S-58183 Linkoping, Sweden

来源：

SCIENCE | 2013年 / 341卷 / 6141期

基金：

欧洲研究理事会; 瑞典研究理事会;

关键词：

KINASE INHIBITORS; PROTEIN STABILITY; PHASE-I; CANCER; RESISTANCE; PARP; PHARMACOKINETICS; SUPPRESSION; MECHANISMS; DISCOVERY;

D O I：

10.1126/science.1233606

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：84 / 87

页数：4

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