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Development of VHL-recruiting STING PROTACs that suppress innate immunity

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Abstract

STING acts as a cytosolic nucleotide sensor to trigger host defense upon viral or bacterial infection. While STING hyperactivation can exert anti-tumor effects by increasing T cell filtrates, in other contexts hyperactivation of STING can contribute to autoimmune and neuroinflammatory diseases. Several STING targeting agonists and a smaller subset of antagonists have been developed, yet STING targeted degraders, or PROTACs, remain largely underexplored. Here, we report a series of STING-agonist derived PROTACs that promote STING degradation in renal cell carcinoma (RCC) cells. We show that our STING PROTACs activate STING and target activated/phospho-STING for degradation. Locking STING on the endoplasmic reticulum via site-directed mutagenesis disables STING translocation to the proteasome and resultingly blocks STING degradation. We also demonstrate that PROTAC treatment blocks downstream innate immune signaling events and attenuates the anti-viral response. Interestingly, we find that VHL acts as a bona fide E3 ligase for STING in RCC; thus, VHL-recruiting STING PROTACs further promote VHL-dependent STING degradation. Our study reveals the design and biological assessment of VHL-recruiting agonist-derived STING PROTACs, as well as demonstrates an example of hijacking a physiological E3 ligase to enhance target protein degradation via distinct mechanisms.

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Data and materials availability

All data supporting the findings in this study are available from the corresponding author upon reasonable request.

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Acknowledgements

We thank Liu and James lab members for critical reading of the manuscript and helpful discussions.

Funding

This work is supported by Gabrielle’s Angel Foundation Medical Research Award (PL) and University of North Carolina at Chapel Hill University Cancer Research Fund (PL). This research is based in part upon work conducted using the UNC Proteomics Core Facility, which is supported in part by NCI Center Core Support Grant (2P30CA016086-45) to the UNC Lineberger Comprehensive Cancer Center.

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Author notes

  1. Zhichuan Zhu and Rebecca L. Johnson contributed equally.

Authors and Affiliations

  1. Lineberger Comprehensive Cancer Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Zhichuan Zhu, Zhigang Zhang, Blossom Damania, Lindsey I. James & Pengda Liu

  2. Department of Biochemistry and Biophysics, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Zhichuan Zhu, Guochun Jiang & Pengda Liu

  3. Center for Integrative Chemical Biology and Drug Discovery, Division of Chemical Biology and Medicinal Chemistry, UNC Eshelman School of Pharmacy, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Rebecca L. Johnson & Lindsey I. James

  4. Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Zhigang Zhang & Blossom Damania

  5. UNC Proteomics Core Facility, Department of Pharmacology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Laura E. Herring

  6. UNC HIV Cure Center, Institute of Global Health and Infectious Diseases, The University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Guochun Jiang

  7. University of North Carolina Center for AIDS Research, The University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA

    Blossom Damania

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  1. Zhichuan Zhu
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Contributions

Z.Z., R.L.J., L.I.J. and P.L. conceived the project and designed experiments. Z.Z., R.L.J., Z.Z. and L.E.H performed experiments. Z.Z., Z.Z., L.E.H., G.J. analyzed the data. B.D., L.I.J. and P.L. supervised this study. Z.Z., R.L.J, L.I.J, and P.L. wrote and edited the manuscript.

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Correspondence to Lindsey I. James or Pengda Liu.

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Zhu, Z., Johnson, R.L., Zhang, Z. et al. Development of VHL-recruiting STING PROTACs that suppress innate immunity. Cell. Mol. Life Sci. 80, 149 (2023). https://doi.org/10.1007/s00018-023-04796-7

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