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Cited by 94interferon and immune responsesImmunotherapy and Immune ResponsesCancer Immunotherapy and Biomarkers

Epigenetic state determines the in vivo efficacy of STING agonist therapy

Open access
Falahat, Rana, Berglund, Anders, Perez-Villarroel, Patricio, Putney, Ryan M., Hamaidi, Imene, Kim, Sungjune, Pilon-Thomas, Shari, Barber, Glen N. +1 more·2023·Nature Communications
DOI 10.1038/s41467-023-37217-1PMC PMC10033671
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Abstract

While STING-activating agents have shown limited efficacy in early-phase clinical trials, multiple lines of evidence suggest the importance of tumor cell-intrinsic STING function in mediating antitumor immune responses. Although STING signaling is impaired in human melanoma, its restoration through epigenetic reprogramming can augment its antigenicity and T cell recognition. In this study, we show that reversal of methylation silencing of STING in murine melanoma cell lines using a clinically available DNA methylation inhibitor can improve agonist-induced STING activation and type-I IFN induction, which, in tumor-bearing mice, can induce tumor regression through a CD8 + T cell-dependent immune response. These findings not only provide mechanistic insight into how STING signaling dysfunction in tumor cells can contribute to impaired responses to STING agonist therapy, but also suggest that pharmacological restoration of STING signaling through epigenetic reprogramming might improve the therapeutic efficacy of STING agonists.

AI snapshot

Written from the paper’s full text.

TL;DR

Reversal of promoter hypermethylation with 5AZADC restores STING expression and agonist-induced type-I IFN/MHC-I in murine melanoma, and combining 5AZADC with ADU-S100 improves tumor control in STING-deficient and STING-intact hosts. The combination’s efficacy depends on CD8+ T cells and is lost in STING-deficient tumors.

Key findings

  • In B16-F10 and Yumm1.7 melanoma cells, 5AZADC reduced STING promoter methylation and restored STING protein; with ADU-S100, IFN-β increased up to 46-fold (p < 0.0001) and MHC class I increased. In B16-ISG cells, IFN-β increased 36-fold and MHC class I 4.4-fold (p < 0.0001), but not in B16-ISG-STINGKO cells.
  • siRNA depletion of DNMT3A and DNMT3B, but not DNMT1, reconstituted STING mRNA/protein and increased ADU-S100-induced CXCL10 up to 2.5-fold and IFN-β up to 3.6-fold (p < 0.0001).
  • In STINGgt/gt mice, 5AZADC + ADU-S100 significantly delayed B16-F10 (p < 0.05 on day 22), Yumm1.7 (p < 0.0001 on day 21), and B16-ISG (p < 0.0001 on day 21) tumor growth, but not B16-ISG-STINGKO tumors; combination increased intratumoral Ifnb1 and H2-k1 (p < 0.0001) and Cxcl10 2.6-fold (p < 0.0001).
  • CD8+ T cell depletion abrogated combination efficacy (p < 0.001 on day 22), while CD4+ depletion did not; combination increased CD8+ TIL frequency (p < 0.05), CXCR3+ CD8+ T cells, and less exhausted LAG-3/PD-1 phenotype, and in C57BL/6 mice was more effective than ADU-S100 alone.

Methods

  • Murine melanoma lines B16-F10, Yumm1.7, B16-ISG, B16-ISG-STINGKO and human A375/SK-MEL-28 cells; in vitro 5AZADC 0.1–1 μM, ADU-S100 10 µg/ml, siRNA against DNMT1/3A/3B; methylation profiling with Infinium Mouse Methylation BeadChip; ELISA, immunoblot, qPCR, and flow cytometry.
  • In vivo subcutaneous tumor models in STINGgt/gt or C57BL/6 mice; intratumoral ADU-S100 50 μg and/or 5AZADC 0.1 mg/kg or PBS; tumor volume measured; T cell depletion with anti-CD8 or anti-CD4; cBioPortal metastatic melanoma dataset (n = 21) for correlation analysis.
  • Statistics used two-tailed t tests, one-way or two-way ANOVA, and Pearson correlation; significance defined as p < 0.05.

Limitations

  • Stated: additional experiments are needed to fully understand factors contributing to systemic activation; optimal dosing levels and therapeutic schedules remain to be identified; future work is needed in solid tumors beyond melanoma and with T cell/CAR T-cell therapies.
  • Evident from the text: findings are preclinical murine models with small group sizes (e.g., n = 3–7 per group) and no human clinical validation; STINGgt/gt hosts lack host STING/T cell STING, so some immune contributions may differ from intact hosts.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.1038/s41467-023-37217-1

Cites

64 of 64 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

49 of 67 cited sources checked · 23 supported · 3 flagged · accuracy not yet measured

  • Premisecitation

    STING-activating agents have shown limited efficacy in early-phase clinical trials in cancer patients, either as monotherapy or in combination with immune checkpoint inhibitors.

    “early-phase clinical trials with these agents in cancer patients, either as monotherapy or in combination with immune checkpoint inhibitors, have so far shown lack of efficacy26–28.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [26] Harrington, K. et al. Preliminary results of the first-in-human (FIH) study of MK-1454, an agonist of stimulator of interferon genes (STING), as monotherapy or in combination with pembrolizumab (pembro) in patients with advanced solid tumors or lymphomas. Ann. Oncol. 29, viii712 (2018). source

      The Abstract reports no complete or partial responses with MK-1454 monotherapy, but six partial responses among 25 evaluated patients receiving MK-1454 plus pembrolizumab. Its conclusion calls the combination’s efficacy “encouraging,” contrary to the claim that the combination showed a lack of efficacy. (LLM, conf=0.88)

      Checked by a reasoning model — may be wrong.

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    • cites [27] van der Zanden, S. Y., Luimstra, J. J., Neefjes, J., Borst, J. & Ovaa, H. Opportunities for small molecules in cancer immunotherapy. Trends Immunol. 41, 493–511 (2020). source
    • cites [28] F. Meric-Bernstam et al. Phase I Dose-Escalation Trial of MIW815 (ADU-S100), an Intratumoral STING Agonist, in Patients with Advanced/Metastatic Solid Tumors or Lymphomas. Clin. Cancer Res. 28, 677–688 (2022). source

      The Translational Relevance section describes limited clinical activity with single-agent MIW815, and the Efficacy section reports one confirmed partial response (2.1% overall response rate). This trial did not test MIW815 with a checkpoint inhibitor; the Discussion instead describes emerging clinical data suggesting greater activity with combined treatment. The source therefore supports the monotherapy part, but not the claim that combination trials have shown a lack of efficacy. (LLM, conf=0.88)

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  • Premisecitation

    Clinical trials in cancer patients currently testing STING activating drugs as monotherapy or in combination with immune checkpoint inhibitors have so far provided little, if any, evidence of objective responses.

    “However, clinical trials in cancer patients currently testing STING activating drugs as a monotherapy or in combination with immune checkpoint inhibitors have so far provided little, if any, evidence of objective responses26–28.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [26] Harrington, K. et al. Preliminary results of the first-in-human (FIH) study of MK-1454, an agonist of stimulator of interferon genes (STING), as monotherapy or in combination with pembrolizumab (pembro) in patients with advanced solid tumors or lymphomas. Ann. Oncol. 29, viii712 (2018). source

      The Results (abstract, p. viii712) report no complete or partial responses with MK-1454 monotherapy, but six partial responses among 25 patients (24%) receiving MK-1454 plus pembrolizumab. Those objective responses contradict the claim that the trials provided little, if any, evidence of objective responses, although the monotherapy result is consistent with it. (LLM, conf=0.87)

      Checked by a reasoning model — may be wrong.

      Experimental — accuracy not yet measured
    • cites [27] van der Zanden, S. Y., Luimstra, J. J., Neefjes, J., Borst, J. & Ovaa, H. Opportunities for small molecules in cancer immunotherapy. Trends Immunol. 41, 493–511 (2020). source
    • cites [28] F. Meric-Bernstam et al. Phase I Dose-Escalation Trial of MIW815 (ADU-S100), an Intratumoral STING Agonist, in Patients with Advanced/Metastatic Solid Tumors or Lymphomas. Clin. Cancer Res. 28, 677–688 (2022). source

      The Efficacy section reports one confirmed partial response among patients receiving MIW815 alone, an objective response rate of 2.1%, supporting the claim of little response to monotherapy. The paper does not establish that objective responses have been similarly scarce across STING-agonist trials combined with checkpoint inhibitors; its Discussion instead notes emerging clinical data showing greater activity with combination treatment. Page numbers were not available in our copy. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Premisecitation

    Tumor cell-intrinsic STING activity is an important contributor to antitumor immune responses.

    “recent studies have provided compelling evidence that tumor cell-intrinsic STING activity is also an important contributor34–36.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [34] Wang, Z. et al. cGAS/STING axis mediates a topoisomerase II inhibitor–induced tumor immunogenicity. J. Clin. Investig. 129, 4850–4862 (2019). source

      The available passage (article header, no page number) gives the title, which links cGAS/STING to drug-induced tumor immunogenicity, but it contains no findings about tumor cell-intrinsic STING activity or antitumor immune responses. Searches for those terms returned only the header, so the decisive article text appears to be missing from our copy. (LLM, conf=0.70)

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    • cites [35] Kitajima, S. et al. Suppression of STING associated with LKB1 loss in KRAS-driven lung cancer. Cancer Discov. 9, 34–45 (2019). source
    • cites [36] Mahadevan, N. R. et al. Intrinsic immunogenicity of small cell lung carcinoma revealed by its cellular plasticity. Cancer Discov. 11, 1952–1969 (2021). source
  • Premisecitation

    Activation of STING signaling in melanoma cells induces CXCL10 and CCL5.

    “Another aspect of the activation of STING signaling in melanoma cells is the downstream induction of CXCL10 and CCL529.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [29] Falahat, R. et al. STING signaling in melanoma cells shapes antigenicity and can promote antitumor T-cell activity. Cancer Immunol. Res. 7, 1837–1848 (2019). source

      The Discussion (page unavailable) reports that STING activation by 2′3′-cGAMP induced CXCL10 in a subset of human melanoma cell lines. The Results section on intact STING signaling also reports CXCL10 induction, but our copy does not report induction of CCL5; searches for CCL5 and RANTES found no supporting passage. (LLM, conf=0.88)

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  • Premisecitation

    Together with CXCL9, CXCL10 is a dominant chemokine for the recruitment of tumor-specific CD8+ T cells into tumors.

    “Together with CXCL9, another CXCR3-binding chemokine, CXCL10 is a dominant chemokine for the recruitment of tumor-specific CD8+ T cells into the tumors31,32.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites [31] Mikucki, M. et al. Non-redundant requirement for CXCR3 signalling during tumoricidal T-cell trafficking across tumour vascular checkpoints. Nat. Commun. 6, 7458 (2015). source
    • cites [32] Dangaj, D. et al. Cooperation between constitutive and inducible chemokines enables T cell engraftment and immune attack in solid tumors. Cancer Cell 35, 885–900.e810 (2019). source

      The Results and Discussion link CXCL9 to CD8+ T-cell infiltration and T-cell recruitment. The Discussion says CXCL10 may have a complementary role, but that CXCL10 expression does not correlate closely with CD8+ infiltration and may not translate to effective infiltration. The paper therefore does not support calling CXCL10 a dominant recruitment chemokine alongside CXCL9. Page numbers are unavailable in our copy. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Classified as probably not a substantial claim. 21
  • May combine two claims. 11
  • The quote only partly states this claim. 1

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The importance of IFN-β production by cancer cells in the recruitment and activation of Batf3-dependent dendritic cells has been demonstrated in different settings.

    “The importance of IFN-β production by cancer cells in the recruitment and activation of Batf3-dependent dendritic cells has been also demonstrated in different settings47,49.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [47] Vanpouille-Box, C. et al. DNA exonuclease Trex1 regulates radiotherapy-induced tumour immunogenicity. Nat. Commun. 8, 1–15 (2017). source
    • cites [49] Duong, E. et al. Type I interferon activates MHC class I-dressed CD11b+ conventional dendritic cells to promote protective anti-tumor CD8+ T cell immunity. Immunity 55, 308–323.e309 (2022). source

      The Results section on IFN-β addition (page unavailable) reports activation and increased infiltration of ISG+ dendritic cells in Batf3−/− mice, which lack Batf3-dependent DC1s. The Discussion identifies these cells as CD11b+ DCs and says IFN-β can drive anti-tumor responses without DC1s. Thus, although the paper links tumor-derived type I interferon to dendritic-cell activation, its demonstrated mechanism is contrary to the claim that it recruits and activates Batf3-dependent dendritic cells. (LLM, conf=0.85)

      Checked by a reasoning model — may be wrong.

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  • May combine two claims.

    Premisecitation

    Activation of STING signaling in human melanoma cell lines can increase their antigenicity by upregulating MHC class I molecules, leading to more effective immune recognition and antigen presentation to tumor-infiltrating lymphocytes.

    “we previously reported that activation of STING signaling in human melanoma cell lines can increase their antigenicity by upregulating MHC class I molecules, leading to more effective immune recognition and antigen presentation to tumor-infiltrating lymphocytes (TIL)29.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [29] Falahat, R. et al. STING signaling in melanoma cells shapes antigenicity and can promote antitumor T-cell activity. Cancer Immunol. Res. 7, 1837–1848 (2019). source

  • May combine two claims.

    Premisecitation

    STING is a major pathway for the detection of immunogenic tumors and initiation of a spontaneous T cell response.

    “the stimulator of interferon genes (STING) has been shown to be a major pathway for the detection of immunogenic tumors and initiation of a spontaneous T cell response18–21.”
    Quote in the paperSubstantial claim2 of 4 sources checked
    • cites [18] Woo, S.-R. et al. STING-dependent cytosolic DNA sensing mediates innate immune recognition of immunogenic tumors. Immunity 41, 830–842 (2014). source
    • cites [19] Sun, L., Wu, J., Du, F., Chen, X. & Chen, Z. J. Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science 339, 786–791 (2013). source
    • cites [20] Ishikawa, H. & Barber, G. N. STING is an endoplasmic reticulum adaptor that facilitates innate immune signalling. Nature 455, 674–678 (2008). source

      The paper’s Abstract reports STING-mediated responses to intracellular DNA and pathogens, but does not address detection of immunogenic tumors or initiation of a spontaneous T-cell response. Searches for those terms and for tumor, cancer, and antitumor immunity found no passage supporting the attributed claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [21] Burdette, D. L. et al. STING is a direct innate immune sensor of cyclic di-GMP. Nature 478, 515–518 (2011). source

      The paper’s Abstract describes STING as a sensor of cyclic dinucleotides and an adaptor in the interferon response to cytosolic DNA. Searches for immunogenic tumors, tumor detection, and spontaneous T-cell responses found no passage in our copy addressing those parts of the claim. (LLM, conf=0.76)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    DNA demethylating agents can induce a type I IFN response in cancer cells through de-repression of endogenous retroviruses and activation of the MDA5-MAVS pathway.

    “multiple studies have shown that DNA demethylating agents among other epigenetic modulators can induce a type I IFN response in cancer cells through the de-repression of endogenous retroviruses (ERVs) and activation of the MDA5-MAVS pathway38–40.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [38] Chiappinelli, K. B. et al. Inhibiting DNA methylation causes an interferon response in cancer via dsRNA including endogenous retroviruses. Cell 162, 974–986 (2015). source
    • cites [39] Topper, M. J. et al. Epigenetic therapy ties MYC depletion to reversing immune evasion and treating lung cancer. Cell 171, 1284–1300.e1221 (2017). source

      The Introduction and Results section on immune-related genes state that DNA methyltransferase inhibition can induce an IFNα/β response involving dsRNA from endogenous retroviruses in cancer cells. The cited paper does not identify activation of the MDA5–MAVS pathway as the mechanism; searches for MDA5 and MAVS found no supporting passage. Page numbers were not available in our copy. (LLM, conf=0.87)

      Experimental — accuracy not yet measured
    • cites [40] Sheng, W. et al. LSD1 ablation stimulates anti-tumor immunity and enables checkpoint blockade. Cell 174, 549–563.e519 (2018). source

      The Discussion (Intracellular dsRNA homeostasis; page not available) reports that DNMT inhibitors increase ERV transcription and IFN activation. The Results (TLR3 and MDA5 sense dsRNA accumulation; page not available) show MDA5–MAVS involvement after LSD1 inhibition, not after treatment with DNA demethylating agents. Thus the source does not establish that full pathway for those agents. (LLM, conf=0.88)

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  • Classified as probably not a substantial claim.

    Premisecitation

    Intratumoral presence of CD8+ T cells is linked to a type-I interferon transcriptional signature.

    “multiple clinical observations have linked intratumoral presence of CD8+ T cells to a type-I interferon (IFN) transcriptional signature14–16.”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [14] Fuertes, M. B. et al. Host type I IFN signals are required for antitumor CD8+ T cell responses through CD8α+ dendritic cells. J. Exp. Med. 208, 2005–2016 (2011). source
    • cites [15] Ulloa-Montoya, F. et al. Predictive gene signature in MAGE-A3 antigen-specific cancer immunotherapy. J. Clin. Oncol. 31, 2388–2395 (2013). source

      The Discussion links tumor CD8 markers with IFNG-regulated genes and downstream targets of STAT1 and IRF1. That supports an association between a CD8-related tumor signature and interferon signaling, but the paper identifies IFNG rather than a type-I interferon signature; it does not establish the claimed type-I specificity. (LLM, conf=0.85)

      Experimental — accuracy not yet measured
    • cites [16] Sistigu, A. et al. Cancer cell–autonomous contribution of type I interferon signaling to the efficacy of chemotherapy. Nat. Med. 20, 1301 (2014). source

  • Classified as probably not a substantial claim. May combine two claims.

    Premisecitation

    CXCL10 and CCL5 are dominant mediators for recruitment of CXCR3+ and CCR5+ tumor-specific T lymphocytes into tumors, and their intratumoral expression correlates with favorable clinical outcomes in melanoma.

    “These chemokines have been identified as the dominant mediators for the recruitment of CXCR3+ and CCR5+ tumor-specific T lymphocytes into the tumors and their intratumoral expression correlated with favorable clinical outcomes in patients with melanoma30–32.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [30] Wolchok, J. D. et al. Overall survival with combined nivolumab and ipilimumab in advanced melanoma. N. Engl. J. Med. 377, 1345–1356 (2017). source

      The Discussion reports increased CXCL10 expression in the tumor microenvironment with combination treatment, but does not connect it to T-cell recruitment or favorable clinical outcomes. Searches for CCL5, CCR5, CXCR3, and intratumoral chemokine expression did not find passages addressing those claims. The paper’s survival analyses concern treatment and PD-L1 expression, not the claimed chemokine–outcome correlation. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites [31] Mikucki, M. et al. Non-redundant requirement for CXCR3 signalling during tumoricidal T-cell trafficking across tumour vascular checkpoints. Nat. Commun. 6, 7458 (2015). source

      The Introduction links tumor CXCL9/CXCL10 expression and CCR5 ligands, including CCL5, with T-cell infiltration and favorable clinical outcomes. But the Results identify CXCR3 as required for T-cell entry into melanoma tumors and report that loss of CCR5 did not impair entry. The paper therefore does not support calling CCL5–CCR5 a dominant recruitment pathway alongside CXCL10–CXCR3. (LLM, conf=0.88)

      Experimental — accuracy not yet measured
    • cites [32] Dangaj, D. et al. Cooperation between constitutive and inducible chemokines enables T cell engraftment and immune attack in solid tumors. Cancer Cell 35, 885–900.e810 (2019). source

      The Introduction links CXCL10 and CCL5 to T-cell infiltration in melanoma and describes the importance of CXCR3 and CCR5 on tumor-infiltrating lymphocytes. However, the Results identify CCL5 and CXCL9—not CXCL10—as the key pair associated with T-cell engraftment and longer survival in melanoma. Section labels are available in our copy, but page numbers are not. (LLM, conf=0.88)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    Type-I IFNs are important in the initiation of spontaneous and iatrogenic antitumor immune T cell responses.

    “several preclinical studies have confirmed the important function of type-I IFNs in the initiation of spontaneous and iatrogenic antitumor immune T cell responses”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [16] Sistigu, A. et al. Cancer cell–autonomous contribution of type I interferon signaling to the efficacy of chemotherapy. Nat. Med. 20, 1301 (2014). source
    • cites [17] Diamond, M. S. et al. Type I interferon is selectively required by dendritic cells for immune rejection of tumors. J. Exp. Med. 208, 1989–2003 (2011). source

      The Results and Discussion show that type I IFN is needed early for tumor-specific cytotoxic T-cell priming and identify its role in spontaneous CD8+ T-cell responses (page not provided). The paper discusses administered IFN-α in cancer treatment, but treats its immune mechanism as uncertain and calls for future study of therapeutic responses; it does not establish initiation of iatrogenic antitumor T-cell responses. (LLM, conf=0.86)

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  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    A 12-chemokine gene expression signature uncovers the presence of tumor-localized tertiary lymphoid structures, which positively correlate with overall survival in certain patients with metastatic melanoma.

    “they are also included in a 12-chemokine gene expression signature that we have shown to uncover the presence of the tumor-localized, tertiary lymphoid structures, which are found to positively correlate with overall survival in certain patients with metastatic melanoma33.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [33] Messina, J. L. et al. 12-Chemokine gene signature identifies lymph node-like structures in melanoma: potential for patient selection for immunotherapy? Sci. Rep. 2, 765 (2012). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    In host dendritic cells, STING-mediated IFN-β induction is essential for their activation and the cross-priming of cytotoxic T cells.

    “In host dendritic cells, STING-mediated IFN-β induction has been well characterized and shown to be essential for their activation and the cross-priming of cytotoxic T cells22,23.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [22] Corrales, L. et al. Direct activation of STING in the tumor microenvironment leads to potent and systemic tumor regression and immunity. Cell Rep. 11, 1018–1030 (2015). source

      The Results report STING-dependent IFN-β production and activation markers in dendritic cells, while the Introduction describes type I IFN signaling as critical for tumor-specific T-cell priming and dendritic-cell cross-presentation. However, the paper does not establish that IFN-β induction itself is essential for dendritic-cell activation or cross-priming; those are more specific causal claims than the cited passages support. Page numbers are not available in our copy. (LLM, conf=0.84)

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    • cites [23] Fu, J. et al. STING agonist formulated cancer vaccines can cure established tumors resistant to PD-1 blockade. Sci. Transl. Med. 7, 283ra252 (2015). source

      The Results section (‘Synthetic CDN derivatives have potent human immunostimulatory capacity’; page unavailable) reports that STING agonists stimulated type I interferon production and activation markers in dendritic cells. The Discussion (page unavailable) says the mechanism by which the vaccine activates recruited dendritic cells remains to be investigated. The paper does not establish that IFN-β induction is essential for dendritic-cell activation or for cross-priming cytotoxic T cells. (LLM, conf=0.87)

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  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Recent studies have indicated that STING agonist-induced T cell death depends on the type and doses of STING agonists.

    “Although it was proposed that STING agonist can induce T cell death through the activation of proapoptotic transcriptional programs58, recent studies have indicated that this effect depends on the type and doses of STING agonists59,60.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [59] Imanishi, T. et al. Reciprocal regulation of STING and TCR signaling by mTORC1 for T-cell activation and function. Life Sci. alliance 2, e201800282 (2019). source
    • cites [60] Benoit-Lizon, I. et al. CD4 T cell-intrinsic STING signaling controls the differentiation and effector functions of TH1 and TH9 cells. J. Immunother. cancer 10, e003459 (2022). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    CXCL10 has been shown to promote effector T cell generation and function.

    “Additionally, CXCL10 has been shown to promote effector T cell generation and function56,57.”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites [56] Dufour, J. H. et al. IFN-γ-inducible protein 10 (IP-10; CXCL10)-deficient mice reveal a role for IP-10 in effector T cell generation and trafficking. J. Immunol. 168, 3195–3204 (2002). source
    • cites [57] Chow, M. T. et al. Intratumoral activity of the CXCR3 chemokine system is required for the efficacy of anti-PD-1 therapy. Immunity 50, 1498–1512.e1495 (2019). source

      The Discussion (page unavailable) states that CD11b+ dendritic cell-derived CXCL10 is essential for optimal generation of CD4+ Th1 responses, supporting the generation part. The Introduction discusses T-cell function for the CXCR3 chemokine system generally, but does not establish that CXCL10 specifically promotes effector T-cell function. (LLM, conf=0.83)

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  • Probably not a substantial claim: may not pass S1 (asserted), S3 (load-bearing).

    Premisecitation

    It was proposed that STING agonist can induce T cell death through activation of proapoptotic transcriptional programs.

    “Although it was proposed that STING agonist can induce T cell death through the activation of proapoptotic transcriptional programs58, recent studies have indicated that this effect depends on the type and doses of STING agonists59,60.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [58] Larkin, B. et al. Cutting edge: activation of STING in T cells induces type I IFN responses and cell death. J. Immunol. 199, 397–402 (2017). source

  • Classified as probably not a substantial claim.

    Premisecitation

    The ability to target and restore STING signaling in tumor cells using epigenetic modulators provides a rationale to explore whether epigenetic reprogramming of tumor cell-intrinsic STING activity can improve the therapeutic efficacy of STING agonists.

    “Despite this common suppression, the ability to target and restore STING signaling in tumor cells using epigenetic modulators35,37 provides a rationale to explore whether epigenetic reprogramming of tumor cell-intrinsic STING activity can improve the therapeutic efficacy of STING agonists.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [35] Kitajima, S. et al. Suppression of STING associated with LKB1 loss in KRAS-driven lung cancer. Cancer Discov. 9, 34–45 (2019). source
    • cites [37] Falahat, R. et al. Epigenetic reprogramming of tumor cell–intrinsic STING function sculpts antigenicity and T cell recognition of melanoma. Proc. Natl Acad. Sci. USA 118, e2013598118 (2021). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Initiation of an adaptive immune response to radiation therapy has been linked to STING-mediated IFN-β induction by dendritic cells.

    “Similarly, initiation of an adaptive immune response to radiation therapy has been linked to STING-mediated IFN-β induction by dendritic cells45.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [45] Deng, L. et al. STING-dependent cytosolic DNA sensing promotes radiation-induced type I interferon-dependent antitumor immunity in immunogenic tumors. Immunity 41, 843–852 (2014). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Tumor-associated tertiary lymphoid structures are emerging as a powerful prognostic marker in melanoma and a number of other cancer types.

    “Also, as a member of the 12-chemokine gene signature classifier33, its expression has been correlated with the presence of tumor-associated tertiary lymphoid structures50–52 which are rapidly emerging as a powerful prognostic marker in melanoma and a number of other cancer types53–55.”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [53] Cabrita, R. et al. Tertiary lymphoid structures improve immunotherapy and survival in melanoma. Nature 577, 561–565 (2020). source
    • cites [54] Helmink, B. A. et al. B cells and tertiary lymphoid structures promote immunotherapy response. Nature 577, 549–555 (2020). source

      The section “B cells prognostic in TCGA analysis” describes existing evidence for a potential prognostic role for TLSs in melanoma and other cancer types. It also reports improved survival for a melanoma group with higher B-cell expression, but that is not a direct measure of TLSs. The source therefore supports the emerging prognostic association, not the stronger description of TLSs as a powerful prognostic marker across those cancers. (LLM, conf=0.83)

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    • cites [55] Meylan, M. et al. Tertiary lymphoid structures generate and propagate anti-tumor antibody-producing plasma cells in renal cell cancer. Immunity 55, 527–541.e525 (2022). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    CXCL10 expression has been correlated with the presence of tumor-associated tertiary lymphoid structures.

    “Also, as a member of the 12-chemokine gene signature classifier33, its expression has been correlated with the presence of tumor-associated tertiary lymphoid structures50–52 which are rapidly emerging as a powerful prognostic marker in melanoma and a number of other cancer types53–55.”
    Quote in the paperNot a substantial claim3 of 3 sources checked
    • cites [50] Coppola, D. et al. Unique ectopic lymph node-like structures present in human primary colorectal carcinoma are identified by immune gene array profiling. Am. J. Pathol. 179, 37–45 (2011). source
    • cites [51] Prabhakaran, S. et al. Evaluation of invasive breast cancer samples using a 12-chemokine gene expression score: correlation with clinical outcomes. Breast Cancer Res. 19, 1–11 (2017). source

      The Background identifies CXCL10 as one of the 12 chemokines in the signature, and the Results show that breast tumors with high signature scores had ectopic lymph node-like structures. The paper correlates the combined 12-chemokine score with these structures; it does not establish the correlation for CXCL10 expression individually. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites [52] Li, R. et al. The 12-CK score: global measurement of tertiary lymphoid structures. Front. Immunol. 12, 694079 (2021). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Two recent reports have shown that T cell-intrinsic STING signaling could control differentiation and effector functions of CD8+ and CD4+ T cells.

    “Also, two recent reports have shown that T cell-intrinsic STING signaling could control differentiation and effector functions of CD8+ and CD4+ T cells60,61.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [60] Benoit-Lizon, I. et al. CD4 T cell-intrinsic STING signaling controls the differentiation and effector functions of TH1 and TH9 cells. J. Immunother. cancer 10, e003459 (2022). source
    • cites [61] Li, W. et al. cGAS-STING–mediated DNA sensing maintains CD8+ T cell stemness and promotes antitumor T cell therapy. Sci. Transl. Med. 12, eaay9013 (2020). source

  • May combine two claims.

    Premisecitation

    In human melanoma cell lines, STING-mediated IFN-β induction can augment surface expression of MHC class I and render them better targets for cytotoxic CD8+ T cell recognition and killing.

    “Indeed, we have previously shown that in human melanoma cell lines STING-mediated IFN-β induction can augment their surface expression of MHC class I and therefore render them better targets for cytotoxic CD8+ T cell recognition and killing29,37.”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [29] Falahat, R. et al. STING signaling in melanoma cells shapes antigenicity and can promote antitumor T-cell activity. Cancer Immunol. Res. 7, 1837–1848 (2019). source
    • cites [37] Falahat, R. et al. Epigenetic reprogramming of tumor cell–intrinsic STING function sculpts antigenicity and T cell recognition of melanoma. Proc. Natl Acad. Sci. USA 118, e2013598118 (2021). source

  • Classified as probably not a substantial claim.

    Premisecitation

    Direct activation of the host STING pathway through intratumoral injections of synthetic cyclic dinucleotides is an attractive therapeutic strategy in multiple preclinical cancer models.

    “Direct activation of the host STING pathway through intratumoral injections of synthetic cyclic dinucleotides has shown to be an attractive therapeutic strategy in multiple preclinical cancer models22,25,42.”
    Quote in the paperSubstantial claim3 of 3 sources checked
    • cites [22] Corrales, L. et al. Direct activation of STING in the tumor microenvironment leads to potent and systemic tumor regression and immunity. Cell Rep. 11, 1018–1030 (2015). source
    • cites [25] Sivick, K. E. et al. Magnitude of therapeutic STING activation determines CD8+ T cell-mediated anti-tumor immunity. Cell Rep. 25, 3074–3085.e3075 (2018). source

      The available indexed passages contain a table of contents and reference links, but no substantive text about treatment outcomes. Searches for intratumoral synthetic cyclic dinucleotides, multiple tumor models, and tumor regression found no passage that establishes the claimed preclinical therapeutic promise. (LLM, conf=0.72)

      Experimental — accuracy not yet measured
    • cites [42] Nicolai, C. J. et al. NK cells mediate clearance of CD8+ T cell–resistant tumors in response to STING agonists. Sci. Immunol. 5, eaaz2738 (2020). source

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Two recent publications describe a novel role for cancer cells and their type I IFN production in priming CD8+ T cell responses through MHC class I dressing of dendritic cells.

    “This notion is further supported by two recent publications describing a novel role for cancer cells and their type I IFN production in priming CD8+ T cell responses through MHC class I dressing of dendritic cells49,62.”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [49] Duong, E. et al. Type I interferon activates MHC class I-dressed CD11b+ conventional dendritic cells to promote protective anti-tumor CD8+ T cell immunity. Immunity 55, 308–323.e309 (2022). source
    • cites [62] MacNabb, B. W. et al. Dendritic cells can prime anti-tumor CD8+ T cell responses through major histocompatibility complex cross-dressing. Immunity 55, 2206–2208 (2022). source

  • May combine two claims.

    Findingown data

    In 5AZADC-pretreated B16-ISG cells, stimulation with STING agonist increased IFN-β induction 36-fold (p < 0.0001) and MHC class I surface expression 4.4-fold (p < 0.0001), but not in 5AZADC-pretreated B16-ISG-STINGKO cells.

    “we found a 36-fold increase in induction of IFN-β (p < 0.0001) (Supplementary Fig. 3b) as well as a 4.4-fold increase in surface expression of MHC class I (p < 0.0001) (Supplementary Fig. 3c, d) in 5AZADC-pretreated B16-ISG cells in response to stimulation with the STING agonist, but not in 5AZADC-pretreated B16-ISG-STINGKO cells.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: B16-ISG-STINGKO cells are STING-deficient.
    • Rests on an assumption: ADU-S100 is a STING agonist.

  • May combine two claims.

    Findingown data

    5AZADC alone induced IFN-β and upregulated MHC class I in B16-F10 and Yumm1.7 cells, though to a lesser extent.

    “Although to a lesser extent, we also observed 5AZADC-mediated induction of IFN-β and upregulation of MHC class I in both B16-F10 and Yumm1.7 cell lines (Fig. 2f–h).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    On day 14, STING agonist treatment significantly increased the frequency and total number of CD8+ T cells in B16-ISG tumors compared to control, but not in B16-ISG-STINGKO tumors.

    “flow cytometry analysis of the TIL on day 14 indicated a significant increase in the frequency and a total number of CD8+ T cells in STING agonist-treated B16-ISG tumors compared to their control group (Supplementary Fig. 1c, d) but not in B16-ISG-STINGKO tumors.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: STINGgt/gt mice have defective host STING signaling, allowing isolation of tumor cell-intrinsic STING effects.
    • Rests on an assumption: B16-ISG-STINGKO cells are STING-deficient.
    • Rests on an assumption: ADU-S100 is a STING agonist.

  • The quote only partly states this claim.

    Findingown data

    By day 20, ADU-S100 lost the ability to maintain tumor control in B16-ISG tumors in STINGgt/gt mice.

    “by day 20, ADU-S100 lost the ability to maintain tumor control in B16-ISG tumors (Fig. 1a, b).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: STINGgt/gt mice have defective host STING signaling, allowing isolation of tumor cell-intrinsic STING effects.
    • Rests on an assumption: ADU-S100 is a STING agonist.

  • May combine two claims.

    Interpretationown data

    Epigenetic reprogramming can restore melanoma-intrinsic STING signaling defects and prime therapeutic responses to STING agonism in vivo.

    “Collectively, these results show that epigenetic reprogramming can restore melanoma-intrinsic STING signaling defects and therefore prime therapeutic responses to STING agonism in vivo.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: STINGgt/gt mice have defective host STING signaling, allowing isolation of tumor cell-intrinsic STING effects.
    • Rests on an assumption: B16-ISG-STINGKO cells are STING-deficient.
    • Rests on an assumption: ADU-S100 is a STING agonist.

  • May combine two claims.

    Interpretationown data

    Although melanoma-intrinsic STING activity can mediate antitumor immune responses at early stages of tumor growth (1-2 weeks), it is insufficient to mediate long-lasting tumor control in response to STING agonist treatment.

    “Collectively, these results indicate that although melanoma-intrinsic STING activity can mediate antitumor immune responses at the early stages of tumor growth (1–2 weeks), it is insufficient to mediate long-lasting tumor control in response to treatment with a STING agonist.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: STINGgt/gt mice have defective host STING signaling, allowing isolation of tumor cell-intrinsic STING effects.
    • Rests on an assumption: B16-ISG-STINGKO cells are STING-deficient.
    • Rests on an assumption: ADU-S100 is a STING agonist.

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Successful depletions were confirmed by flow cytometry analysis of splenocytes.

    “Successful depletions were confirmed by flow cytometry analysis of splenocytes (Supplementary Fig. 5).”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Data are representative of two independent experiments.

    “Data are shown as the mean ± SEM and are representative of two independent experiments.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    STING-functional tumors can initiate a CXCL10/CXCR3 axis and subsequently guide trafficking and entry of effector T cells into tumor sites.

    “These data suggest that STING-functional tumors can initiate a CXCL10/CXCR3 axis and subsequently guide trafficking and entry of effector T cells into the tumor sites.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductnone given

    Images are representative of three independent experiments.

    “Images are representative of three independent experiments.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Study design and conductown data

    Cellular depletions were confirmed by flow cytometry of splenocytes.

    “Cellular depletions were confirmed by flow cytometry of splenocytes (Supplementary Fig. 5).”
    Quote in the paperNot a substantial claim
31 of 31

  • The paper relies on this without support.

    STINGgt/gt mice have defective host STING signaling, allowing isolation of tumor cell-intrinsic STING effects.

    “we injected STINGgt/gt mice subcutaneously with B16-ISG or B16-ISG-STINGKO murine melanoma cells”
    “This approach allowed us to separate the effect of tumor cell-intrinsic STING activity from that driven by the host immune system.”

  • The paper relies on this without support.

    B16-ISG-STINGKO cells are STING-deficient.

    “B16-ISG-STINGKO murine melanoma cells”
    “ADU-S100 treatment did not impact B16-ISG-STINGKO tumor growth.”

  • The paper relies on this without support.

    ADU-S100 is a STING agonist.

    “treated them intratumorally with either the STING agonist ADU-S100 or PBS as vehicle control”

  • The paper relies on this without support.

    The murine melanoma models used in this study are predictive of human melanoma for STING agonist therapy.

    “In this study, we investigated this rationale in vivo using multiple approaches with murine melanoma models in both STING-deficient and STING-sufficient hosts.”
    “insights from our study provide a framework to design proper clinical treatment modalities with appropriate patient selection in melanoma and perhaps other solid tumors.”
4 of 4

Cites

Flagged first, then by citations

Research works printed but not linked: 1

  1. Restifo, N. P. et al. Loss of functional beta 2 -microglobulin in metastatic melanomas from five patients receiving immunotherapy. J. Natl Cancer Inst. 88, 100–108 (1996). · DOI

Cited by

Sorted by citations

Related

Authors
Falahat, R., Berglund, A., Perez-Villarroel, P., Putney, R. M., Hamaidi, I., Kim, S., Pilon-Thomas, S., Barber, G. N., & Mulé, J. J.
Affiliations
  • Moffitt Cancer Center (US)
  • University of Miami (US)
Corresponding author
Mulé, James J.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2023-03-22
Volume
14
Issue
1
Pages
1573-1573
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-023-37217-1
PMC
PMC10033671
OpenAlex
W4353087862
Cited by
94
References
64
Citations by year
2026: 26 · 2025: 35 · 2024: 29 · 2023: 4
FWCI
8.22
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
U.S. Department of Health & Human Services | NIH | National Cancer Institute (P30 CA076292, P50 CA168536)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
  • Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
  • Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
Keywords
STING agonist, epigenetic reprogramming, DNA methylation, melanoma, type I interferon, CD8+ T cells, antitumor immunity, DNA methylation inhibitors, tumor regression
MeSH terms
Animals, Antineoplastic Agents, Humans, Immunity, Interferon Type I, Melanoma, Epigenesis, Genetic, Mice

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-19.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-19.

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