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Universal STING mimic boosts antitumour immunity via preferential activation of tumour control signalling pathways

Nature Nanotechnology (2024)Cite this article

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Abstract

The efficacy of STING (stimulator of interferon genes) agonists is due to various factors, primarily inefficient intracellular delivery, low/lack of endogenous STING expression in many tumours, and a complex balance between tumour control and progression. Here we report a universal STING mimic (uniSTING) based on a polymeric architecture. UniSTING activates STING signalling in a range of mouse and human cell types, independent of endogenous STING expression, and selectively stimulates tumour control IRF3/IFN-I pathways, but not tumour progression NF-κB pathways. Intratumoural or systemic injection of uniSTING-mRNA via lipid nanoparticles (LNPs) results in potent antitumour efficacy across established and advanced metastatic tumour models, including triple-negative breast cancer, lung cancer, melanoma and orthotopic/metastatic liver malignancies. Furthermore, uniSTING displays an effective antitumour response superior to 2′3′-cGAMP and ADU-S100. By favouring IRF3/IFN-I activity over the proinflammatory NF-κB signalling pathway, uniSTING promotes dendritic cell maturation and antigen-specific CD8+ T-cell responses. Extracellular vesicles released from uniSTING-treated tumour cells further sensitize dendritic cells via exosome-containing miRNAs that reduced the immunosuppressive Wnt2b, and a combination of LNP-uniSTING-mRNA with α-Wnt2b antibodies synergistically inhibits tumour growth and prolongs animal survival. Collectively, these results demonstrate the LNP-mediated delivery of uniSTING-mRNA as a strategy to overcome the current STING therapeutic barriers, particularly for the treatment of multiple cancer types in which STING is downregulated or absent.

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Fig. 1: LNP-uniSTING-mRNA induced constitutive STING activation and EV-mediated crosstalk between tumour cells and DCs.
Fig. 2: Characterization of tetramer-based uniSTING as a universal STING agonist independent of cGAMP or endogenous STING.
Fig. 3: Cytosolic delivery of uniSTING-mRNA based on LNPs intratumourally inhibits tumour growth.
Fig. 4: Systemic uniSTING treatment exerts potent antitumour effects on orthotopic/metastatic tumours.
Fig. 5: Exosomal miRNAs derived from uniSTING-treated tumour cells potentiate DC function by blocking Wnt2b signalling.
Fig. 6: α-Wnt2b antibody enhances in vivo antitumour activity of STING activation.

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Data availability

The MicroRNA Data Integration Portal (mirDIP) was used to identify gene targets for exosomal miRNAs and can be accessed at http://ophid.utoronto.ca/mirDIP. The gene set database Hallmarks (h.all.v6.1.symbols.gmt) from the Molecular Signatures Database (MSigDB) was used in the analysis. All raw sequencing data and associated processed data files that support the findings of this study have been deposited in the Gene Expression Omnibus under accession code GSE253724 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE253724). Source data are available for Figs. 2d–g,i,j, 3b,d–q, 4b,d,f,g, 5a–g,i,j and 6b,c,e–h and Supplementary Figs. 1c, 2a, 6b–d, 8c,d, 9 and 13d–f in the associated source data file. Source data are provided with this paper.

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Acknowledgements

This work was supported by innovation grants (RX03202109 and RX03222104) from the Eshelman Institute for Innovation (to R.L.), a developmental grant (MCR0634222) from UCRF (to R.L.) and research grants from the NIH (R01EB032865 to R.L. and R35-CA232109 and R01-AI029564 to J.P.-Y.T.). E.M. was supported by T32-CA196589.

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  1. These authors contributed equally: Ying Wang, Sirui Li.

Authors and Affiliations

  1. Division of Chemical Biology and Medicinal Chemistry, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Ying Wang, Ellie McCabe, Lillian Zhang, Andrew M. Withrow & Rihe Liu

  2. Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Sirui Li, Jenny P.-Y. Ting & Rihe Liu

  3. Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Sirui Li & Jenny P.-Y. Ting

  4. Children’s Cancer and Blood Foundation Laboratories, Departments of Pediatrics, and Cell and Developmental Biology, Drukier Institute for Children’s Health, Meyer Cancer Center, Weill Cornell Medicine, New York, NY, USA

    Mengying Hu

  5. State Key Laboratory of Biocontrol, School of Ecology, Sun Yat-sen University, Guangzhou, China

    Yuchen Yang

  6. Carolina Center for Genome Sciences, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA

    Rihe Liu

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Contributions

Y.W., S.L., J.P.-Y.T. and R.L. conceived and designed the research. R.L. designed the uniSTING. Y.W. and S.L. performed plasmid construction, protein expression, mRNA synthesis, LNP generation, and all the tissue culture and in vivo animal experiments. M.H. performed RNA-seq and GSEA analysis and participated in the design of EV-related experiments. Y.Y. performed raw data alignments and analysis from RNA-seq results. E.M., L.Z. and A.M.W. helped with the preparation of mRNA and LNPs. Y.W. and M.H. performed the statistical analysis. With input from J.P.-Y.T. and S.L., Y.W. and R.L. analysed all the data and wrote the manuscript.

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Correspondence to Jenny P.-Y. Ting or Rihe Liu.

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Wang, Y., Li, S., Hu, M. et al. Universal STING mimic boosts antitumour immunity via preferential activation of tumour control signalling pathways. Nat. Nanotechnol. (2024). https://doi.org/10.1038/s41565-024-01624-2

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