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

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CETSA and thermal proteome profiling strategies for target identification and drug discovery of natural products
Phytomedicine ( IF 6.7 ) Pub Date : 2023-05-20 , DOI: 10.1016/j.phymed.2023.154862
Yanbei Tu ₁ , Lihua Tan ₂ , Hongxun Tao ₃ , Yanfang Li ₄ , Hanqing Liu ₁

Affiliation

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.

中文翻译：

CETSA 和热蛋白质组分析策略用于天然产物的目标识别和药物发现

背景

在药物开发的各个阶段监测目标参与对于基于天然产物 (NP) 的药物发现和开发至关重要。2013 年开发的细胞热转移分析 (CETSA) 是一种新颖的、广泛适用的、无标记的生物物理分析，基于配体诱导的靶蛋白热稳定原理，可以直接评估生理相关环境中的药物-靶点参与，包括完整的细胞、细胞裂解物和组织。本综述旨在概述CETSA的工作原理及其衍生策略及其在蛋白质靶标验证、靶标识别和NPs药物先导发现方面的最新进展。

方法

使用 Web of Science 和 PubMed 数据库进行了一项基于文献的调查。审查和讨论了所需的信息，以突出 CETSA 衍生策略在 NP 研究中的重要作用。

结果

经过近十年的升级和演变，CETSA主要发展为三种格式：用于目标验证的经典Western blotting（WB）-CETSA，用于无偏蛋白质组目标识别的热蛋白质组分析（TPP，也称为MS-CETSA）和高通量 (HT)-CETSA，用于药物命中发现和先导化合物优化。重要的是，强调和讨论了各种 TPP 方法在生物活性 NPs 目标发现中的应用可能性，包括 TPP-温度范围（TPP-TR）、TPP-化合物浓度范围（TPP-CCR）、二维 TPP（ 2D-TPP)、细胞表面-TPP (CS-TPP)、简化的 TPP (STPP)、二维凝胶电泳中基于热稳定性位移的荧光差异 (TS-FITGE) 和沉淀物支持的 TPP (PSTPP)。此外，关键优势，