Phenotypic proteomic profiling identifies a landscape of targets for circadian clock–modulating compounds

Sandipan Ray, Radoslaw Lach, Kate J Heesom, Utham K Valekunja, Vesela Encheva, Ambrosius P. Snijders, Akhilesh B Reddy

Research output: Contribution to journalArticle (Academic Journal)peer-review

10 Citations (Scopus)
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Determining the exact targets and mechanisms of action of drug molecules that modulate circadian rhythms is critical to develop novel compounds to treat clock-related disorders. Here, we have used phenotypic proteomic profiling (PPP) to systematically determine molecular targets of four circadian period–lengthening compounds in human cells. We demonstrate that the compounds cause similar changes in phosphorylation and activity of several proteins and kinases involved in vital pathways, including MAPK, NGF, B-cell receptor, AMP-activated protein kinases (AMPKs), and mTOR signaling. Kinome profiling further indicated inhibition of CKId, ERK1/2, CDK2/7, TNIK, and MST4 kinases as a common mechanism of action for these clock-modulating compounds. Pharmacological or genetic inhibition of several convergent kinases lengthened circadian period, establishing them as novel circadian targets. Finally, thermal stability profiling revealed binding of the compounds to clock regulatory kinases, signaling molecules, and ubiquitination proteins. Thus, phenotypic proteomic profiling defines novel clock effectors that could directly inform precise therapeutic targeting of the circadian system in humans.
Original languageEnglish
Number of pages17
JournalLife Science Alliance
Early online date2 Dec 2019
Publication statusE-pub ahead of print - 2 Dec 2019


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