CETSA and thermal proteome profiling strategies for target identification and drug discovery of natural products

Yanbei Tu; Lihua Tan; Hongxun Tao; Yanfang Li; Hanqing Liu

doi:10.1016/j.phymed.2023.154862

CETSA and thermal proteome profiling strategies for target identification and drug discovery of natural products

Phytomedicine. 2023 Jul 25:116:154862. doi: 10.1016/j.phymed.2023.154862. Epub 2023 May 20.

Authors

Yanbei Tu¹, Lihua Tan², Hongxun Tao³, Yanfang Li⁴, Hanqing Liu⁵

Affiliations

¹ School of Pharmacy, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
² State Key Laboratory of Quality Research in Chinese Medicine, Institute of Chinese Medical Sciences, University of Macau, Taipa, Macao SAR 999078, China.
³ School of Food and Biological Engineering, Jiangsu University, Zhenjiang, Jiangsu 212013, China.
⁴ School of Chemical Engineering, Sichuan University, Chengdu, Sichuan 610065, China. Electronic address: lyf471@vip.163.com.
⁵ School of Pharmacy, Jiangsu University, Zhenjiang, Jiangsu 212013, China. Electronic address: hanqing@ujs.edu.cn.

PMID: 37216761
DOI: 10.1016/j.phymed.2023.154862

Abstract

Background: Monitoring target engagement at various stages of drug development is essential for natural product (NP)-based drug discovery and development. The cellular thermal shift assay (CETSA) developed in 2013 is a novel, broadly applicable, label-free biophysical assay based on the principle of ligand-induced thermal stabilization of target proteins, which enables direct assessment of drug-target engagement in physiologically relevant contexts, including intact cells, cell lysates and tissues. This review aims to provide an overview of the work principles of CETSA and its derivative strategies and their recent progress in protein target validation, target identification and drug lead discovery of NPs.

Methods: A literature-based survey was conducted using the Web of Science and PubMed databases. The required information was reviewed and discussed to highlight the important role of CETSA-derived strategies in NP studies.

Results: After nearly ten years of upgrading and evolution, CETSA has been mainly developed into three formats: classic Western blotting (WB)-CETSA for target validation, thermal proteome profiling (TPP, also known as MS-CETSA) for unbiased proteome-wide target identification, and high-throughput (HT)-CETSA for drug hit discovery and lead optimization. Importantly, the application possibilities of a variety of TPP approaches for the target discovery of bioactive NPs are highlighted and discussed, including TPP-temperature range (TPP-TR), TPP-compound concentration range (TPP-CCR), two-dimensional TPP (2D-TPP), cell surface-TPP (CS-TPP), simplified TPP (STPP), thermal stability shift-based fluorescence difference in 2D gel electrophoresis (TS-FITGE) and precipitate supported TPP (PSTPP). In addition, the key advantages, limitations and future outlook of CETSA strategies for NP studies are discussed.

Conclusion: The accumulation of CETSA-based data can significantly accelerate the elucidation of the mechanism of action and drug lead discovery of NPs, and provide strong evidence for NP treatment against certain diseases. The CETSA strategy will certainly bring a great return far beyond the initial investment and open up more possibilities for future NP-based drug research and development.

Keywords: Cellular thermal shift assay; Natural products; Target discovery; Target validation; Thermal proteome profiling.

Publication types

Review

MeSH terms

Biological Products* / pharmacology
Drug Delivery Systems
Drug Discovery / methods
High-Throughput Screening Assays / methods
Proteome* / metabolism

Substances

Proteome
Biological Products