Key points are not available for this paper at this time.
Abstract The CEllular Thermal Shift Assay (CETSA) technology is a powerful way to assess target engagement across various matrixes. By the introduction of the high-throughput format (CETSA Navigate HT), this technology offers a reliable high-throughput screening method in natural cellular environments that can successfully be applied in compound screening for hit identification and confirmation already during lead generation. Due to its broad applicability to native, full length protein targets in the live cellular environment, without the need for expensive and time-consuming protein and/or compound labelling efforts, CETSA Navigate HT may be regarded as a new paradigm for high-throughput screening and actives generation. As a case study to demonstrate the applicability of CETSA Navigate HT in hit identification, we have applied the technology for primary screening of a 12000 compound library for CDK4, a protein targeted in several cancers. Hits identified from this screen are privileged from the onset by binding to the target in a physiologically relevant cellular matrix and in a binding-mode agnostic way. This is in contrast to traditional screening methods targeting activity or in vitro interactions where cellular activity is addressed in later stages. Data from follow up studies using functional assays will be presented as well as mass spectrometry based CETSA selectivity profiling and comparison to commercially available de novo CDK4/6 inhibitors. Citation Format: Helena Almqvist, Victoria Brehmer, Laurence Arnold, Tomas Friman, Stina Lundgren, Daniel Martinez Molina. CETSA Navigate HT: A new paradigm for high-throughput screening abstract. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts) ; 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84 (6Suppl): Abstract nr 3096.
Almqvist et al. (Fri,) studied this question.