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Blockade of USP14 potentiates type I interferon signaling and radiation-induced antitumor immunity via preventing IRF3 deubiquitination

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Abstract

Purpose

The adaptive immune responses induced by radiotherapy has been demonstrated to largely rely on STING-dependent type I interferons (IFNs) production. However, irradiated tumor cells often fail to induce dendritic cells (DCs) to produce type I IFNs. Hence, we aim to uncover the limitation of STING-mediated innate immune sensing following radiation, and identify efficient reagents capable to rescue the failure of type I IFNs induction for facilitating radiotherapy.

Methods

A targeted cell-based phenotypic screening was performed to search for active molecules that could elevate the production of type I IFNs. USP14 knockout or inhibition was assayed for IFN production and the activation of STING signaling in vitro. The mechanisms of USP14 were investigated by western blot and co-immunoprecipitation in vitro. Additionally, combinational treatments with PT33 and radiation in vivo and in vitro models were performed to evaluate type I IFNs responses to radiation.

Results

PT33 was identified as an enhancer of STING agonist elicited type I IFNs production to generate an elevated and durable STING activation profile in vitro. Mechanistically, USP14 inhibition or deletion impairs the deubiquitylation of K63-linked IRF3. Furthermore, blockade of USP14 with PT33 enhances DC sensing of irradiated-tumor cells in vitro, and synergizes with radiation to promote systemic antitumor immunity in vivo.

Conclusion

Our findings reveal that USP14 is one of the major IFN production suppressors and impairs the activation of IRF3 by removing the K63-linked ubiquitination of IRF3. Therefore, blockage of USP14 results in the gain of STING signaling activation and radiation-induced adaptive immune responses.

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Data availability

All data are available within the article, supplementary information, or available from the corresponding author upon reasonable request.

Abbreviations

USP14 :

Ubiquitin-specific protease 14

IFNs :

Interferons

DCs :

Dendritic cells

STING :

Stimulator of interferon genes

TBK1 :

TANK-binding kinase 1

IRF3 :

Interferon regulatory factor 3

TME :

Tumor microenvironment

cGAMP :

Cyclic GMP-AMP

cGAS :

cGAMP synthase

DUBs :

Deubiquitinases

RIG-I :

Retinoic acid-inducible gene I

USPs :

Ubiquitin-specific proteases

HCV-1 :

Herpes simplex virus type 1

BMDCs :

Bone marrow-derived dendritic cells

CDNs :

Cyclic dinucleotides

KO :

Knockout

OE :

Overexpression

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Funding

This project was supported by the National Natural Science Foundation of China (NSFC) (No. 81803054 and 81973174), Guangdong Basic and Applied Basic Research Foundation (No. 2021A1515012496 and 2022A1515012221).

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Author notes

  1. Weijian Wu and Huiying Xu contributed equally to this work.

Authors and Affiliations

  1. School of Pharmaceutical Sciences, Sun Yat-sen University, Guangzhou, 510006, China

    Weijian Wu, Huiying Xu, Chenyang Liao, Youqiao Wang, Ruirui Wu, Jiaxin Wu, Yunzhi Li, Chaoying Jin, Yuxuan Zhao & Xianzhang Bu

  2. Department of Radiation Oncology, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China

    Xin Yue

  3. Institute of Precision Medicine, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, 510080, China

    Xin Yue

  4. State Key Laboratory of Chemical Oncogenomics, Guangdong Provincial Key Laboratory of Chemical Genomics, Peking University Shenzhen Graduate School, Shenzhen, 518055, China

    Wenlv Zheng, Junmin Quan & Xianzhang Bu

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  1. Weijian Wu
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Contributions

Study concept and design: X.Y. and X.B.; Performed in vitro functional studies and analyzed data: W.W., H.X., C.L., Y.W., and W.Z.; Performed animal experiments and analyzed data: X.Y. and W.W.; Generated critical cellular tools: H.X., W.W., R.W., C.J. and J.W.; Performed mechanistical studies and analyzed data: X.Y., W.W., Y.L., Y.Z. and H.X.; Statistical analysis: X.Y., X.B.; Supervision of data: X.Y., X.B. and J.Q.; Analysis and interpretation of data and writing of the manuscript: X.B. and X.Y.. All authors have read and approved the manuscript.

Corresponding authors

Correspondence to Xin Yue or Xianzhang Bu.

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Animal experiments were approved by the Institutional Ethics Committee for Clinical Research and Animal Trials of the First Affiliated Hospital, Sun Yat-sen University (SYSUFAH). All the experiments were conducted in the Laboratory Animal Center of Sun Yat-sen University.

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Wu, W., Xu, H., Liao, C. et al. Blockade of USP14 potentiates type I interferon signaling and radiation-induced antitumor immunity via preventing IRF3 deubiquitination. Cell Oncol. 45, 1347–1361 (2022). https://doi.org/10.1007/s13402-022-00724-2

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