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bioRxiv · 10.1101/2025.08.08.669362

ZYS-1 is not an ADAR1 inhibitor

Smoak, C. N.·Gardner, E. N.·Chua, R. N.·Cottrell, K. A.
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Abstract

Adenosine deaminase acting on RNA 1 (ADAR1) edits double-stranded RNA (dsRNA) substrates by the deamination of adenosine to inosine in a process known as A-to-I editing. Modulation of ADAR1 expression and editing activity has previously been described to play a role in cancer development and progression, with upregulation of ADAR1 being observed in a range of cancers. Further, depletion of ADAR1 leads to increased sensing of endogenous dsRNAs by dsRNA sensors in cell lines that require ADAR1 for survival, which are termed ADAR1-dependent. The activation of these sensors induces downstream production of type I interferons as well as translational inhibition and apoptosis. Therefore, ADAR1 is a promising oncologic therapeutic target. Recently, the small molecule ZYS-1 has been developed and presented as a direct inhibitor of ADAR1. We performed a series of in vitro and cellular experiments to validate the efficacy and specificity of ZYS-1 as an ADAR1 inhibitor. Evaluating the effect of ZYS-1 on cell viability revealed it to be equally cytotoxic to both ADAR1-dependent and ADAR1-independent cell lines, as well as wildtype and ADAR1 knockout cells. Moreover, ZYS-1 treatment had little effect on activation of PKR or induction of IFN stimulated genes. Importantly, treatment with ZYS-1 did not reduce cellular A-to-I editing for several known ADAR1 editing sites, and did not inhibit in vitro A-to-I editing by recombinant ADAR1. Together, these data indicate that ZYS-1 is not a selective inhibitor of ADAR1.

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Smoak, C. N., Gardner, E. N., Chua, R. N., Cottrell, K. A.. 2025-08-08. ZYS-1 is not an ADAR1 inhibitor. https://doi.org/10.1101/2025.08.08.669362

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