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Rapid-kinetics degron benchmarking reveals off-target activities and mixed agonism-antagonism of MYB inhibitors

Harada, T.; Perez, M. W.; Kalfon, J.; Dievenich Braes, F.; Batley, R.; Eagle, K.; Nabet, B.; Leifer, B.; Kruell, J.; Paralkar, V. R.; Stegmaier, K.; Koehler, A. N.; Orkin, S. H.; Pimkin, M.

2023-04-07 molecular biology
10.1101/2023.04.07.536032 bioRxiv
Show abstract

Attenuating aberrant transcriptional circuits holds great promise for the treatment of numerous diseases, including cancer. However, development of transcriptional inhibitors is hampered by the lack of a generally accepted functional cellular readout to characterize their target specificity and on-target activity. We benchmarked the direct gene-regulatory signatures of six agents reported as inhibitors of the oncogenic transcription factor MYB against targeted MYB degradation in a nascent transcriptomics assay. The inhibitors demonstrated partial specificity for MYB target genes but displayed significant off-target activity. Unexpectedly, the inhibitors displayed bimodal on-target effects, acting as mixed agonists-antagonists. Our data uncover unforeseen agonist effects of small molecules originally developed as TF inhibitors and argue that rapid-kinetics benchmarking against degron models should be used for functional characterization of transcriptional modulators.

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