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Cited by 200Cancer Immunotherapy and BiomarkersCancer, Hypoxia, and MetabolismImmune cells in cancer

Hypoxia induces HIF1α-dependent epigenetic vulnerability in triple negative breast cancer to confer immune effector dysfunction and resistance to anti-PD-1 immunotherapy

Open access
Ma, Shijun, Zhao, Yue, Lee, Wee Chyan, Ong, Li-Teng, Lee, Puay Leng, Jiang, Zemin, Oguz, Gokce, Niu, Zhitong +11 more·2022·Nature Communications
DOI 10.1038/s41467-022-31764-9PMC PMC9287350
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Abstract

The hypoxic tumor microenvironment has been implicated in immune escape, but the underlying mechanism remains elusive. Using an in vitro culture system modeling human T cell dysfunction and exhaustion in triple-negative breast cancer (TNBC), we find that hypoxia suppresses immune effector gene expression, including in T and NK cells, resulting in immune effector cell dysfunction and resistance to immunotherapy. We demonstrate that hypoxia-induced factor 1α (HIF1α) interaction with HDAC1 and concurrent PRC2 dependency causes chromatin remolding resulting in epigenetic suppression of effector genes and subsequent immune dysfunction. Targeting HIF1α and the associated epigenetic machinery can reverse the immune effector dysfunction and overcome resistance to PD-1 blockade, as demonstrated both in vitro and in vivo using syngeneic and humanized mice models. These findings identify a HIF1α-mediated epigenetic mechanism in immune dysfunction and provide a potential strategy to overcome immune resistance in TNBC.

AI snapshot

Written from the paper’s full text.

TL;DR

Hypoxia in triple-negative breast cancer suppresses T and NK effector gene expression via HIF1α-dependent recruitment of HDAC1 and pre-existing PRC2, producing chromatin remodeling and resistance to anti-PD-1 therapy. Targeting HIF1α or the associated epigenetic machinery restores immune effector function and sensitizes TNBC to PD-1 blockade in vitro and in mouse models.

Key findings

  • In TCGA basal TNBC samples (n = 98 in Fig. 1a), hypoxia gene signature expression was inversely correlated with immune effector genes (IFNG, TNF, GZMB, PRF1), IFNγ-responsive genes, and TIL-associated genes; in human TNBC samples (n = 30), HIF1α-high tumor areas had reduced CD8+ T cell infiltration and IFNγ expression.
  • Hypoxia (1% O2) suppressed IFNγ, TNFα, and granzyme B in CD8+ T cells and NK cells, and induced TIM-3/TIGIT; hypoxia plus TNBC coculture increased PD-1+TIM-3+ double-positive CD8+ T cells to 30–40% versus 10–20% in resting T cells.
  • Hypoxia increased HIF1α and HDAC1 occupancy at IFNG and TNF promoters, reduced H3K27ac, and increased H3K27me3; HIF1α co-immunoprecipitated with HDAC1, and HDAC1 also interacted with SUZ12. HDAC inhibitor entinostat, EZH2 inhibitor EPZ6438, HIF1α inhibitors PX478/digoxin, or CRISPR depletion of HDAC1/EZH2 restored effector expression.
  • In 4T1 syngeneic and humanized NIKO/LM2 mouse models, anti-PD-1 alone had minimal effect, but anti-PD-1 combined with PX478 or entinostat induced tumor regression, suppressed lung metastasis, and improved survival; depletion of T or NK cells largely abolished combination efficacy.

Methods

  • Human T and NK cells were isolated from healthy donor PBMC, expanded, and cultured under normoxia (20% O2) or hypoxia (1% O2), alone or cocultured with TNBC lines BT549, MDA-MB-231, or MDA-MB-231-LM2; assays included RNA-seq, flow cytometry, RT-qPCR, ChIP-qPCR, co-immunoprecipitation, CRISPR knockdown, and luciferase-based cytotoxicity.
  • TCGA breast cancer data were analyzed by PAM50 subtype (basal/TNBC n = 98; HER2+ n = 58; Luminal A n = 231; Luminal B n = 129); immunofluorescence used human TNBC samples (n = 30) and mouse 4T1 tumors.
  • In vivo work used BALB/c mice bearing orthotopic 4T1-Luc tumors and humanized NOD/SCID/IL2Rg−/− (NIKO) mice reconstituted with human CD34+ cells and engrafted with MDA-MB-231-LM2; treatments included anti-PD-1/Keytruda with PX478, entinostat, or EPZ6438. Statistics used ANOVA, t-tests, and log-rank tests; RNA-seq DEG thresholds were |logFC| > 1 and adjusted P < 0.01.

Limitations

  • The paper states that PX478 and epigenetic drugs have additional effects on other tumor microenvironment elements such as myeloid cells and macrophages, and that EPZ6438 could not achieve satisfactory solubility in different formulations, likely explaining its weaker in vivo effects.
  • The paper notes that HIF1α-independent induction of TIM-3 and TIGIT remains to be explored as a possible additional strategy to prevent T cell exhaustion.
  • Evident from the text: the mechanistic evidence relies heavily on in vitro healthy-donor human immune cells and mouse/humanized models, with no clinical trial validation; TCGA findings are correlative, and the 1% O2 system may not fully capture heterogeneous tumor hypoxia.

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

Integrity signals

Registry

Crossref · 10.1038/s41467-022-31764-9

Cites

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Statistics

Nothing to recompute.

Claims the paper makes

0 of 56 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    The hypoxia gene signature is inversely correlated with immune effector genes (IFNG, TNF, GZMB, PRF1), IFNγ-responsive genes, and TIL-associated genes in TNBC (n = 99) in the TCGA dataset.

    “data analysis of The Cancer Genome Atlas (TCGA) dataset11 identified that hypoxic gene signature, which includes commonly documented hypoxia-responsive genes26, was inversely correlated with immune effector genes (IFNG, TNF, GZMB, PRF1), IFNγ-responsive genes, as well as genes associated with tumor-infiltrating lymphocytes (TILs) in TNBC (n = 99) (also referred as a basal subtype of breast cancer)”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [11] Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature 490, 61–70 (2012). source

      Not checked in the free run.

    • cites [26] Mahara, S. et al. HIFI-alpha activation underlies a functional switch in the paradoxical role of Ezh2/PRC2 in breast cancer. Proc. Natl Acad. Sci. USA 113, E3735–E3744 (2016). source

      Not checked in the free run.

  • Findingown data

    The protocol efficiently differentiates peripheral naive T cells into antigen-specific cytotoxic effector and memory T cells.

    “Therefore, we showed that this protocol could efficiently differentiate the peripheral naive T cells into antigen-specific cytotoxic effector and memory T cells.”
    Quote in the paperNot a substantial claim
  • Findingown data

    Hypoxia caused marked downregulations of IFNγ, TNFα, and granzyme B in CD8+ T cells in both monoculture and coculture conditions.

    “The results showed that hypoxia caused marked downregulations of IFNγ, TNFα, and granzyme B in CD8+ T cells in both monoculture and coculture conditions (Fig. 2f).”
    Quote in the paperSubstantial claim
  • Findingown data

    TNBC patients with higher immune effector gene signatures are associated with prolonged survivals.

    “Conversely, TNBC patients with higher immune effector gene signatures are associated with prolonged survivals (Fig. 1h).”
    Quote in the paperSubstantial claim
  • Findingown data

    In human TNBC, tumor areas with high levels of hypoxia, indicated by HIF1α staining, displayed markedly reduced CD8+ T cell infiltration and IFNγ expression compared to low hypoxic areas.

    “Intratumoral analysis showed that tumor areas with high levels of hypoxia, as indicated by staining of HIF1α, displayed markedly reduced CD8+ T cell infiltration as well as IFNγ expression compared to low hypoxic areas (Fig. 1c, d).”
    Quote in the paperSubstantial claim
  • May combine two claims. 51
  • Classified as probably not a substantial claim. 27
  • The quote only partly states this claim. 1

  • May combine two claims.

    Findingown data

    TNBC patients with higher hypoxia gene signature are associated with worse overall survival and distant metastasis-free survival compared to those with lower expression of the hypoxia gene signature.

    “Kaplan–Meier analysis using public breast cancer meta-analysis database11,30 reveals that TNBC patients with higher hypoxia gene signature are associated with worse disease outcomes, including both overall survival (OS) and distant metastasis-free survival (DMFS), compared to those with lower expression of hypoxia gene signature (Fig. 1h).”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [11] Cancer Genome Atlas Network. Comprehensive molecular portraits of human breast tumours. Nature 490, 61–70 (2012). source

      Not checked in the free run.

    • cites [30] Nagy, A., Munkacsy, G. & Gyorffy, B. Pancancer survival analysis of cancer hallmark genes. Sci. Rep. 11, 6047 (2021). source

      Not checked in the free run.

  • May combine two claims.

    Findingown data

    Hypoxia did not affect CD4+ and CD8+ T cell compositions, nor overall T cell differentiation status, though it slightly decreased the Tem population and increased the Teff population.

    “Hypoxia did not seem to affect the CD4+ and CD8+ T cell compositions (Fig. S2b), nor did it affect the overall T cell differentiation status, though it slightly decreased the Tem population and increased Teff population (Fig. S2d).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The inverse correlation between hypoxia gene signature and immune gene activity was also observed in HER2+ breast cancer but not in luminal breast cancers.

    “Further analysis of other breast cancer subtypes revealed that the inverse correlation between hypoxia gene signature and immune gene activity was also observed in HER2+ breast cancer but not in luminal breast cancers, which were less aggressive and less hypoxic (Fig. 1b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Flow cytometry confirmed that intratumor T cells with higher levels of PIM exhibited lower numbers of CD8+ T cells and reduced IFNγ expression in CD8+ T cells.

    “Flow cytometry analysis also confirmed that intratumor T cells with higher levels of PIM exhibited lower numbers of CD8+ T cells and reduced IFNγ expression in CD8+ T cells (Fig. 1g).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A similar survival association was found in HER2+ breast cancer but not in luminal breast cancers.

    “A similar result was also found in HER2+ breast cancer but not in luminal breast cancers (Fig. S1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In the 4T1 murine breast tumor model, tumor areas with higher PIM exhibited minimum IFNγ and CD8 signals, while lower PIM areas were enriched with IFNγ and CD8 signals.

    “Tumor areas showing higher levels of PIM exhibited minimum staining of IFNγ and CD8 signals, while the tumor areas showing lower levels of PIM were enriched with IFNγ and CD8 signals (Fig. 1f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    IFNG was one of the top genes downregulated in hypoxia and was consistently downregulated in all three donors.

    “Among them, IFNG (IFNγ) was one of the top genes downregulated in hypoxia and was consistently downregulated in all three donors.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Hypoxia treatment resulted in substantial transcriptional changes in T cells, including marked downregulation of a cohort of cytokines and upregulation of genes enriched in the hypoxia pathway.

    “The data analysis showed that hypoxia treatment resulted in substantial transcriptional changes in T cells. These included marked downregulation of a cohort of cytokines expression, one of the critical hallmark features of T cell exhaustion25, accompanied by upregulation of genes enriched in the hypoxia pathway (Fig. 2d, e; twofold change, p < 0.01 as cutoffs).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [25] Collier, J. L., Weiss, S. A., Pauken, K. E., Sen, D. R. & Sharpe, A. H. Not-so-opposite ends of the spectrum: CD8(+) T cell dysfunction across chronic infection, cancer and autoimmunity. Nat. Immunol. 22, 809–819 (2021). source

      Not checked in the free run.

  • May combine two claims.

    Findingown data

    Hypoxia significantly induced TIM-3 and TIGIT protein expression, though it did not affect the classic checkpoint molecules PD-1 and CTLA-4.

    “Interestingly, hypoxia significantly induced TIM-3 and TIGIT protein expression, though it did not affect the classic checkpoint molecules PD-1 and CTLA-4 (Fig. 2f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia-induced suppression of immune effectors occurred across all subsets of CD8+ cells, though it was more obvious in effector subsets (Tem and Teff).

    “Of notice, hypoxia-induced suppression of immune effectors occurred across all the subsets of CD8+ cells, despite more obvious in effector subsets (both Tem and Teff) (Fig. S2e), indicating that hypoxia can induce effector suppression regardless of T cell differentiation.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    HDAC inhibition by ENT or EZH2 inhibition by EPZ reversed the hypoxia-induced changes in H3K27ac and H3K27me3 at IFNG.

    “Consistent with this hypothesis, we see that HDAC inhibition by HDAC inhibitor ENT or EZH2 inhibition by EPZ reversed the hypoxia-induced changes in H3K27ac and H3K27me3 at IFNG (Fig. 3f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    H3K27ac at IFNG and TNF showed decreased enrichment in hypoxia while H3K27me3 enrichment was increased.

    “Corresponding to the increased HDAC1 enrichment in hypoxia, H3K27ac at IFNG and TNF showed decreased enrichment in hypoxia while H3K27me3 enrichment was increased (Fig. 3e and Fig. S3d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia-induced suppression of effector expression and upregulation of TIM-3 was also observed in NK cells isolated from PBMC, though the effect on TIGIT was not so obvious upon coculture with TNBC.

    “Hypoxia-induced suppression of effector expression and upregulation of TIM-3 was also observed in NK cells isolated from PBMC (Fig. S2g, h), though the effect on TIGIT was not so obvious upon coculture with TNBC (Fig. S2h).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    EZH2 and SUZ12 already showed significant enrichment at IFNG in normoxia, indicating a pre-existing PRC2 complex at IFNG, and hypoxia did not further increase their enrichments.

    “Interestingly, EZH2 and its PRC2 partner SUZ12 already showed a significant enrichment at IFNG in normoxia, indicating a pre-existing PRC2 complex at IFNG, and hypoxia did not further increase their enrichments (Fig. 3d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia did not change the bulk expression levels of HDACs, EZH2, SUZ12, and EED, nor the global levels of H3K27me3 and H3K27ac in T and NK cells.

    “Of note, hypoxia did not change the bulk expression levels of HDACs, EZH2, SUZ12, and EED (Fig. S4d), nor the global levels of H3K27me3 and H3K27ac in T and NK cells (Fig. S4d).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CRISPR-mediated depletion of HDAC1 or EZH2 recapitulated the effect of ENT or EPZ and restored effector gene expression in T and NK cells.

    “Further validation using CRISPR-mediated depletion of HDAC1 or EZH2 recapitulated the effect of ENT or EPZ and restored effector gene expression in T and NK cells (Fig. S4a, b, Fig. 3g, and Fig. S4c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia treatment resulted in higher HDAC1 enrichment at both IFNG and TNF, without affecting HDAC2 and HDAC3.

    “Scanning the promoter regions across −1.5 to 1 Kb of TSS of IFNG and TNF showed that although with minimum enrichments of HDACs in normoxia, hypoxia treatment resulted in higher HDAC1 enrichment at both IFNG and TNF, though without affecting HDAC2 and HDAC3 (Fig. 3d and Fig. S3c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Neither hypoxic culture nor the drug treatment significantly affects the viability of T cells within a time course of 6 days.

    “We also found that neither hypoxic culture nor the drug treatment significantly affects the viability of T cells within a time course of 6 days (Fig. S6c).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    HDAC1, but not HIF1α, was also found in SUZ12 pulldown.

    “HDAC1, but not HIF1α, was also found in SUZ12 pulldown (Fig. 4b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In the absence of immune cells, neither hypoxia nor the above drugs alter IRF1 and PD-L1 expression or viability of the two TNBC cell lines.

    “By contrast, in the absence of immune cells, neither hypoxia nor the above drugs altered the IRF1 and PD-L1 expression and the viability of the two TNBC cell lines (Fig. S7g, h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia weakens mouse T cell cytotoxicity and anti-PD-1 response towards 4T1.

    “Hypoxia also weakened the mouse T cell cytotoxicity and anti-PD-1 response towards 4T1”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Hypoxia-induced resistance to anti-PD-1 is reversed by PX478, ENT, and EPZ, as well as knockdown of HIF1α, HDAC1, or EZH2.

    “Indeed, hypoxia-induced resistance to anti-PD-1 was reversed by PX478, ENT, and EPZ, as well as the knockdown of HIF1α, HDAC1, or EZH2 (Fig. 5e).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Combination treatments reverse the hypoxic effect and promote effector T cell infiltration into the hypoxic region in 4T1 tumors.

    “the combination treatments reversed the hypoxic effect and promoted the effector T cell infiltration into the hypoxic region (Fig. 6h, i).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Combinations with ENT or PX478 restore expression of IFNγ and TNFα in CD8+ TILs and blunt the hypoxic effect in 4T1 tumors.

    “but combinations with ENT or PX478 restored the expression of IFNγ and TNFα in CD8+ TILs and blunted the hypoxic effect (Fig. 6j).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    T cell depletion markedly promotes lung metastasis and abolishes drug combination effects on metastasis.

    “T cell depletion also markedly promoted lung metastasis and abolished the drug combination effects on metastasis”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    EPZ combined with anti-PD-1 improves tumor growth and metastasis in 4T1 mice, though less potent than PX478 and ENT.

    “EPZ combined with anti-PD-1 also improved the effect on tumor growth and metastasis, though the effects were less potent than PX478 and ENT (Fig. S9b, c)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Hypoxia induces HIF1α and HDAC1 co-occupancy at effector genes, leading to reductions of H3K27ac and increases of H3K27me3 through a pre-existing PRC2 presence at effector gene loci, and suppressed effector gene expression in both T and NK cells.

    “We found that hypoxia induces HIF1α and HDAC1 co-occupancy at the effector genes, leading to reductions of H3K27ac and increases of H3K27me3 through a pre-existing PRC2 presence effector gene loci, and suppressed effector gene expression in both T and NK cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In both syngeneic and humanized mouse models, PD-1 blockade plus inhibition of HIF1α or HDAC1 is sufficient to increase intratumoral expression of effector genes on CD8+ T cells and NK cells and IFNγ production in tumors that showed a regressive response.

    “In both syngeneic and humanized mouse models, PD-1 blockade plus inhibition of HIF1α or HDAC1 is sufficient to increase intratumoral expression of effector genes on CD8+ T cells and NK cells and IFNγ production in tumors that showed a regressive response.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Combination treatments significantly promote CD8+ T cell infiltration in hypoxic tumor areas and increase IFNγ expression in intratumoral CD8+ T and NK cells.

    “we have validated that the combination treatments significantly promoted CD8+ T cell infiltration in hypoxic tumor areas and increased IFNγ expression in the intratumoral CD8+ T and NK cells.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that lacks effective treatment.

    “Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer that lacks effective treatment1,2.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [1] Bianchini, G., Balko, J. M., Mayer, I. A., Sanders, M. E. & Gianni, L. Triple-negative breast cancer: challenges and opportunities of a heterogeneous disease. Nat. Rev. Clin. Oncol. 13, 674–690 (2016). source

      Not checked in the free run.

    • cites [2] Podo, F. et al. Triple-negative breast cancer: present challenges and new perspectives. Mol. Oncol. 4, 209–229 (2010). source

      Not checked in the free run.

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

    Premisecitation

    Immune checkpoint blockade directed against PD-1 or PD-L1 has shown promising results in TNBC, but the overall response rate is less than 20%.

    “Immune checkpoint blockade (ICB) directed against programmed death-1 (PD-1) or programmed death-ligand 1 (PD-L1) has shown promising results in TNBC4–6, but the overall response rate of ICB is less than 20%7.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [4] Nanda, R. et al. Effect of pembrolizumab plus neoadjuvant chemotherapy on pathologic complete response in women with early-stage breast cancer: an analysis of the ongoing phase 2 adaptively randomized I-SPY2 trial. JAMA Oncol. 6, 676–684 (2020). source

      Not checked in the free run.

    • cites [5] Schmid, P. et al. Pembrolizumab plus chemotherapy as neoadjuvant treatment of high-risk, early-stage triple-negative breast cancer: results from the phase 1b open-label, multicohort KEYNOTE-173 study. Ann. Oncol. 31, 569–581 (2020). source

      Not checked in the free run.

    • cites [6] Schmid, P. et al. Pembrolizumab for early triple-negative breast cancer. N. Engl. J. Med. 382, 810–821 (2020). source

      Not checked in the free run.

    • cites [7] Nanda, R. et al. Pembrolizumab in patients with advanced triple-negative breast cancer: phase Ib KEYNOTE-012 study. J. Clin. Oncol. 34, 2460–2467 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Despite strong immunogenicity and a higher prevalence of tumor-infiltrating lymphocytes in TNBC, patients often progress to metastasis with poor clinical outcomes.

    “Paradoxically, despite strong immunogenicity and a higher prevalence of tumor-infiltrating lymphocytes (TILs) in TNBC3, patients often progress to metastasis with poor clinical outcomes1.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [3] Liu, Z., Li, M., Jiang, Z. & Wang, X. A comprehensive immunologic portrait of triple-negative breast cancer. Transl. oncol. 11, 311–329 (2018). source

      Not checked in the free run.

    • cites [1] Bianchini, G., Balko, J. M., Mayer, I. A., Sanders, M. E. & Gianni, L. Triple-negative breast cancer: challenges and opportunities of a heterogeneous disease. Nat. Rev. Clin. Oncol. 13, 674–690 (2016). source

      Not checked in the free run.

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

    Premisecitation

    Hypoxia is a hallmark of most solid tumors and a common trait of the tumor microenvironment associated with cancer progression and metastasis.

    “Hypoxia is a hallmark of most solid tumors and a common trait of TME that is associated with cancer progression and metastasis8–10.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [8] Chaturvedi, P., Gilkes, D. M., Takano, N. & Semenza, G. L. Hypoxia-inducible factor-dependent signaling between triple-negative breast cancer cells and mesenchymal stem cells promotes macrophage recruitment. Proc. Natl Acad. Sci. USA 111, E2120–E2129 (2014). source

      Not checked in the free run.

    • cites [9] Rankin, E. B. & Giaccia, A. J. Hypoxic control of metastasis. Science 352, 175–180 (2016). source

      Not checked in the free run.

    • cites [10] Pietrobon, V. & Marincola, F. M. Hypoxia and the phenomenon of immune exclusion. J. Transl. Med. 19, 9 (2021). source

      Not checked in the free run.

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

    Premisecitation

    Contradictory reports show a role of hypoxia in potentiating the activity of cytotoxic T cells, though these studies were mainly conducted in mouse models.

    “Nevertheless, there are also contradictory reports showing the role of hypoxia in potentiating the activity of cytotoxic T cells, though these studies were mainly conducted in mouse models22–24.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [22] Doedens, A. L. et al. Hypoxia-inducible factors enhance the effector responses of CD8(+) T cells to persistent antigen. Nat. Immunol. 14, 1173–1182 (2013). source

      Not checked in the free run.

    • cites [23] Palazon, A. et al. An HIF-1alpha/VEGF-A axis in cytotoxic T cells regulates tumor progression. Cancer Cell 32, 669–683 e665 (2017). source

      Not checked in the free run.

    • cites [24] Labiano, S. et al. CD69 is a direct HIF-1alpha target gene in hypoxia as a mechanism enhancing expression on tumor-infiltrating T lymphocytes. Oncoimmunology 6, e1283468 (2017). source

      Not checked in the free run.

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

    Premisecitation

    Tumor-infiltrating T cell terminal exhaustion resulting from chronic and continuous tumor antigen stimulation is the primary factor contributing to immune evasion and resistance to cancer immunotherapy.

    “Tumor-infiltrating T cell terminal exhaustion resulting from chronic and continuous tumor antigen stimulation is the primary factor contributing to immune evasion and resistance to cancer immunotherapy25.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [25] Collier, J. L., Weiss, S. A., Pauken, K. E., Sen, D. R. & Sharpe, A. H. Not-so-opposite ends of the spectrum: CD8(+) T cell dysfunction across chronic infection, cancer and autoimmunity. Nat. Immunol. 22, 809–819 (2021). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Hypoxia has a role in immune exclusion in TNBC and is associated with poor disease outcomes.

    “Together, these findings support that hypoxia has a role in immune exclusion in TNBC and is associated with poor disease outcomes.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Hypoxia triggered an epigenetic remodeling and HDAC1/PRC2 concurrent dependency, resulting in effector gene suppression.

    “Therefore, hypoxia triggered an epigenetic remodeling and HDAC1/PRC2 concurrent dependency, resulting in effector gene suppression.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    HDAC1 interacts with HIF1α and PRC2 after hypoxia to form two distinct repressor complexes that co-occupy the effector gene promoters.

    “These findings suggest that HDAC1 interacts with HIF1α and PRC2 after hypoxia to form two distinct repressor complexes that co-occupy the effector gene promoters.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Targeting HIF1α and the associated epigenetic machinery can reverse immune effector dysfunction and overcome resistance to PD-1 blockade, as demonstrated both in vitro and in vivo using syngeneic and humanized mice models.

    “Targeting HIF1α and the associated epigenetic machinery can reverse the immune effector dysfunction and overcome resistance to PD-1 blockade, as demonstrated both in vitro and in vivo using syngeneic and humanized mice models.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The in vitro culture system models human T cell dysfunction and exhaustion in TNBC.

  • May combine two claims.

    Interpretationown data

    Hypoxia suppresses immune effector gene expression in T and NK cells, resulting in immune effector cell dysfunction and resistance to immunotherapy.

    “Using an in vitro culture system modeling human T cell dysfunction and exhaustion in triple-negative breast cancer (TNBC), we find that hypoxia suppresses immune effector gene expression, including in T and NK cells, resulting in immune effector cell dysfunction and resistance to immunotherapy.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The in vitro culture system models human T cell dysfunction and exhaustion in TNBC.

  • May combine two claims.

    Interpretationown data

    Hypoxia-induced factor 1α (HIF1α) interaction with HDAC1 and concurrent PRC2 dependency causes chromatin remodeling resulting in epigenetic suppression of effector genes and subsequent immune dysfunction.

    “We demonstrate that hypoxia-induced factor 1α (HIF1α) interaction with HDAC1 and concurrent PRC2 dependency causes chromatin remolding resulting in epigenetic suppression of effector genes and subsequent immune dysfunction.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The in vitro culture system models human T cell dysfunction and exhaustion in TNBC.

  • May combine two claims.

    Interpretationreasoning

    Inhibition of HIF1α, HDAC1, or EZH2 in immune cells paradoxically induces PD-L1 and PD-L2 expression in tumor cells, which could weaken anticancer immunity.

    “Moreover, although inhibition of HIF1α, HDAC1, or EZH2 in immune cells can restore immune cytotoxicity, it also paradoxically induces PD-L1 and PD-L2 expression in tumor cells that in turn could weaken anticancer immunity.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    Hypoxia drives the T cell to an exhausted-like state through both HIF1α-dependent and HIF1α-independent mechanisms.

    “Therefore, hypoxia drives the T cell to an exhausted-like state through both HIF1α-dependent and HIF1α-independent mechanisms.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Interferon signaling is of central importance to antitumor immune response.

    “interferon signaling is of central importance to antitumor immune response43.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [43] Benci, J. L. et al. Tumor interferon signaling regulates a multigenic resistance program to immune checkpoint blockade. Cell 167, 1540–1554 e1512 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The 4T1 xenograft tumor is highly hypoxic and resistant to PD-1 blockade treatment.

    “the 4T1 xenograft tumor is also known to be highly hypoxic and resistant to PD-1 blockade treatment44,45.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [44] Cao, Y. et al. Observation of incipient tumor angiogenesis that is independent of hypoxia and hypoxia inducible factor-1 activation. Cancer Res. 65, 5498–5505 (2005). source

      Not checked in the free run.

    • cites [45] Booth, L. et al. HDAC inhibitors enhance neratinib activity and when combined enhance the actions of an anti-PD-1 immunomodulatory antibody in vivo. Oncotarget 8, 90262–90277 (2017). source

      Not checked in the free run.

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

    Premisecitation

    MDA-MB-231-LM2 (LM2) is an ideal xenograft model for studying tumor growth and lung metastasis.

    “which is an ideal xenograft model for studying tumor growth and lung metastasis47.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [47] Minn, A. J. et al. Genes that mediate breast cancer metastasis to lung. Nature 436, 518–524 (2005). source

      Not checked in the free run.

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

    Premisecitation

    Some studies have shown that hypoxia promotes mouse T cell cytotoxicity.

    “While some studies have shown that hypoxia promotes mouse T cell cytotoxicity22,23”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [22] Doedens, A. L. et al. Hypoxia-inducible factors enhance the effector responses of CD8(+) T cells to persistent antigen. Nat. Immunol. 14, 1173–1182 (2013). source

      Not checked in the free run.

    • cites [23] Palazon, A. et al. An HIF-1alpha/VEGF-A axis in cytotoxic T cells regulates tumor progression. Cancer Cell 32, 669–683 e665 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    In the present study, hypoxia suppresses immune effector activity through epigenetic reprogramming, which, together with continuous antigen stimulation, is crucial to developing immune exhaustion and resistance to immunotherapy.

    “In the present study, we demonstrate that hypoxia suppresses immune effector activity through epigenetic reprogramming, which, together with continuous antigen stimulation, is crucial to developing immune exhaustion and resistance to immunotherapy.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The role of hypoxia in modulating cancer immunity and the corresponding mechanism remains unclear.

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

    Premisecitation

    Hypoxia plays a crucial role in tumor growth, metastasis, and drug resistance in solid tumors.

    “Hypoxia plays a crucial role in tumor growth, metastasis, and drug resistance in solid tumors8,9”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [8] Chaturvedi, P., Gilkes, D. M., Takano, N. & Semenza, G. L. Hypoxia-inducible factor-dependent signaling between triple-negative breast cancer cells and mesenchymal stem cells promotes macrophage recruitment. Proc. Natl Acad. Sci. USA 111, E2120–E2129 (2014). source

      Not checked in the free run.

    • cites [9] Rankin, E. B. & Giaccia, A. J. Hypoxic control of metastasis. Science 352, 175–180 (2016). source

      Not checked in the free run.

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

    Premisecitation

    Ectopic HIF1α overexpression in mouse T cells diminishes antitumor activity in an adoptive-transfer model.

    “A recent study has shown that ectopic HIF1α overexpression in mouse T cells diminished the antitumor activity in an adoptive-transfer model50.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [50] Veliça, P. et al. Modified hypoxia-inducible factor expression in CD8+ T cells increases antitumor efficacy. Cancer Immunol. Res. 9, 401–414 (2021). source

      Not checked in the free run.

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

    Premisecitation

    There are opposite reports showing that tumor hypoxia induces mouse T cell exhaustion and resistance to immunotherapy and depletion of HIF1α increases NK cell activity and tumor infiltration.

    “there are also opposite reports showing that tumor hypoxia induces mouse T cell exhaustion and resistance to immunotherapy and depletion of HIF1α increases NK cell activity and tumor infiltration16,17,19,48,49.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [16] Jayaprakash, P. et al. Targeted hypoxia reduction restores T cell infiltration and sensitizes prostate cancer to immunotherapy. J. Clin. Invest. 128, 5137–5149 (2018). source

      Not checked in the free run.

    • cites [17] Ni, J. et al. Single-cell RNA sequencing of tumor-infiltrating NK cells reveals that inhibition of transcription factor HIF-1alpha unleashes NK cell activity. Immunity 52, 1075–1087 e1078 (2020). source

      Not checked in the free run.

    • cites [19] Scharping, N. E. et al. Mitochondrial stress induced by continuous stimulation under hypoxia rapidly drives T cell exhaustion. Nat. Immunol. 22, 205–215 (2021). source

      Not checked in the free run.

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

    Premisecitation

    Current studies have focused on the role of hypoxia-induced metabolic reprogramming in causing T cell dysfunction and exhaustion.

    “Current studies have focused on the role of hypoxia-induced metabolic reprogramming in causing T cell dysfunction and exhaustion16,19,52–55.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [16] Jayaprakash, P. et al. Targeted hypoxia reduction restores T cell infiltration and sensitizes prostate cancer to immunotherapy. J. Clin. Invest. 128, 5137–5149 (2018). source

      Not checked in the free run.

    • cites [19] Scharping, N. E. et al. Mitochondrial stress induced by continuous stimulation under hypoxia rapidly drives T cell exhaustion. Nat. Immunol. 22, 205–215 (2021). source

      Not checked in the free run.

    • cites [52] Scharping, N. E., Menk, A. V., Whetstone, R. D., Zeng, X. & Delgoffe, G. M. Efficacy of PD-1 blockade is potentiated by metformin-induced reduction of tumor hypoxia. Cancer Immunol. Res. 5, 9–16 (2017). source

      Not checked in the free run.

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

    Premisecitation

    Hypoxia and continued antigen stimulation can induce mitochondrial stress in mouse T cells, leading to terminal exhaustion but in a HIF1α-independent manner.

    “a recent study showed that hypoxia and continued antigen stimulation could induce mitochondrial stress in mouse T cells, leading to terminal exhaustion but in a HIF1α-independent manner19.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [19] Scharping, N. E. et al. Mitochondrial stress induced by continuous stimulation under hypoxia rapidly drives T cell exhaustion. Nat. Immunol. 22, 205–215 (2021). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    In both in vitro and in vivo syngeneic and humanized models, targeting the HIF1α-dependent epigenetic mechanism seems sufficient to rescue T cell dysfunction and sensitizes to PD-1 blockade.

    “Importantly, in both in vitro and in vivo syngeneic and humanized models, we show that targeting the HIF1α-dependent epigenetic mechanism seems sufficient to rescue the T cell dysfunction and sensitizes to PD-1 blockade.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationown data

    This combination holds very promising potential in future treatment of metastatic TNBC.

    “which holds very promising potential in the future treatment of metastatic TNBC.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Our in vitro system for studying human immune cell activity shows that hypoxia can induce HIF1α-dependent epigenetic events to directly suppress immune effector expression in both T and NK cells, leading to impaired immune effector dysfunction.

    “Our in vitro system for studying human immune cell activity shows that hypoxia can induce HIF1α-dependent epigenetic events to directly suppress immune effector expression in both T and NK cells, leading to impaired immune effector dysfunction.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Interpretationreasoning

    Targeting this pathway through either genetic depletion or therapeutic intervention may be helpful to overcome immune escape in TNBC where immune checkpoint immunotherapies have limited response.

    “and targeting this pathway through either genetic depletion or therapeutic intervention may be helpful to overcome immune escape in TNBC where immune checkpoint immunotherapies have limited response.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    ENT plus DNA demethylation agent AZA promotes migration of myeloid cells to the metastatic niche to suppress metastasis in different tumor models.

    “in a recent report, ENT plus DNA demethylation agent AZA was found to promote the migration of myeloid cells to the metastatic niche to suppress metastasis in different tumor models56.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [56] Lu, Z. et al. Epigenetic therapy inhibits metastases by disrupting premetastatic niches. Nature 579, 284–290 (2020). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductcitation

    Only immune-humanized mice reconstituted with 20-50% human CD45+ cells were used for tumor engraftment.

    “Immune-humanized mice reconstituted with 20–50% of human CD45+ cells were used for engraftment59.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    All manipulations and procedures with mice were approved by the A*STAR Institutional Animal Care and Use Committee.

    “All manipulations and procedures with mice were approved by Agency for Science, Technology and Research (A*STAR) Institutional Animal Care and Use Committee (IACUC).”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    Human umbilical cord blood was collected with written consent from the donors' guardians and in accordance with the ethical guidelines of KK Women's and Children's Hospital.

    “Human umbilical cord blood was collected from KK Women’s and Children’s Hospital (KKH), with written consent obtained from guardians of donors, and in accordance with the ethical guidelines of KKH.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Drug treatments at indicated doses did not show toxicity to human T or NK cells.

    “Drug treatments at the indicated doses did not show toxicity to the T or NK cells.”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    In the syngeneic 4T1 mouse TNBC model, mice were randomized before drug treatment began.

    “For the drug administration, mice were randomized and drug treatments started 10 days after tumor injection when the tumor volume reached 70–100 mm3.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable), S3 (load-bearing), S4 (based).

    Interpretationreasoning

    Given that hypoxia is a common trait for solid tumors, both the mechanism and the combinational therapy identified are expandable to treatment of other different tumor types in addition to TNBC.

    “given that hypoxia is a common trait for solid tumors, we believe that both the mechanism and the combinational therapy we have identified are expandable to the treatment of other different tumor types in addition to TNBC.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Hypoxia is a common trait for solid tumors.

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

    Study design and conductnone given

    Drug treatments at indicated doses did not show toxicity to mouse T cells.

    “Drug treatments at the indicated doses did not show toxicity to mouse T cells.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable). May combine two claims.

    Interpretationnone given

    Our findings add to fundamental understanding of hypoxia regulation of immune response, which aids in providing alternative strategies for developing therapeutic intervention to treat TNBC.

    “In summary, our findings add to our fundamental understanding of hypoxia regulation of immune response, which aids in providing alternative strategies for developing a therapeutic intervention to treat TNBC.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    In the TNBC-immune-humanized mouse model, mice were randomized before drug treatment began.

    “For the drug administration, mice were randomized and drug treatments started 15 days after tumor injection when the tumor volume reached 70–100 mm3.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Depletion of T or NK cells abolishes the combination effect, supporting that the synergistic effect acts by removing hypoxia-mediated epigenetic suppression of immune effector expression.

    “Moreover, depletion of T or NK cells abolished the combination effect, further supporting our claim that the synergistic effect act by removing the hypoxia-mediated epigenetic suppression of immune effector expression.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductown data

    Depletion of NK and T cells in mice was verified by flow cytometry of peripheral blood mononuclear cells.

    “The depletion of NK or T cells were examined by flow cytometry of PBMC extracted from mouse blood.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). The quote only partly states this claim.

    Study design and conductnone given

    All experiments were conducted in compliance with animal protocols approved by the ASTAR-Biopolis Institutional Animal Care and Use Committee of Singapore.

    “All experiments were conducted in compliance with animal protocols approved by the ASTAR-Biopolis Institutional Animal Care and Use Committee of Singapore.”
    Quote in the paperNot a substantial claim
70 of 70

  • The paper relies on this without support.

    The in vitro culture system models human T cell dysfunction and exhaustion in TNBC.

    “Using an in vitro culture system modeling human T cell dysfunction and exhaustion in triple-negative breast cancer (TNBC)”

  • The paper relies on this without support.

    The fundamental mechanism governing tumor immune resistance remains unanswered.

    “However, the fundamental mechanism governing tumor immune resistance remains unanswered.”

  • The paper relies on this without support.

    Inhibition of IFNγ production and interferon response including PD-L1 is expected to induce resistance to PD-1/PD-L1 blockade.

    “Inhibition of IFNγ production and interferon response including PD-L1 is expected to induce resistance to the PD-1/PD-L1 blockade.”

  • The paper relies on this without support.

    The role of hypoxia in modulating cancer immunity and the corresponding mechanism remains unclear.

    “but its role in modulating cancer immunity and the corresponding mechanism remains unclear.”

  • The authors state this as an assumption.

    PX478 and epigenetic drugs also have additional effects on other elements such as myeloid cells and macrophages in the tumor microenvironment.

    “Admittedly, there is no doubt that PX478 and epigenetic drugs also have additional effects on other elements such as myeloid cells and macrophages in the tumor microenvironment.”

  • The paper relies on this without support.

    Hypoxia is a common trait for solid tumors.

    “given that hypoxia is a common trait for solid tumors”
6 of 6

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Human immune reconstitution assessment is dated at both 10 and 12 weeks

    Minor

    The Results place the post-transplant assessment at ten weeks, but the Methods say mice were bled to determine immune reconstitution at twelve weeks.

  • TNBC sample count differs between text and figure

    Minor

    The Results report 99 TNBC samples for the TCGA analysis, whereas the Fig. 1 caption reports 98 basal TNBC samples for that analysis.

  • Normoxia is labeled as 1% oxygen

    Minor

    The Fig. 3 caption labels 1% O2 as normoxia, while the paper elsewhere defines 1% O2 as hypoxia and 20% O2 as normoxia.

  • Humanized-mouse cell source differs between Results and Methods

    Minor

    The Results say the mice received human fetal liver cells, whereas the Methods say they received human cord blood cells.

  • Mouse T-cell collection is assigned the wrong day

    Minor

    The cells are seeded into coculture on day 4. Collection after another 48 hours would be on day 6, not day 4.

5 of 5

Cites

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Related

Authors
Ma, S., Zhao, Y., Lee, W. C., Ong, L.-T., Lee, P. L., Jiang, Z., Oguz, G., Niu, Z., Liu, M., Goh, J. Y., Wang, W., Bustos, M. A., Ehmsen, S., Ramasamy, A., Hoon, D. S. B., Ditzel, H. J., Tan, E. Y., Chen, Q., & Yu, Q.
Affiliations
  • Agency for Science, Technology and Research (SG)
  • Sixth Affiliated Hospital of Sun Yat-sen University (CN)
  • Odense University Hospital (DK)
  • University of Southern Denmark (DK)
  • Tan Tock Seng Hospital (SG)
Corresponding author
  • Chen, Qingfeng
  • Yu, Qiang
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2022-07-15
Volume
13
Issue
1
Pages
4118-4118
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-022-31764-9
PMC
PMC9287350
OpenAlex
W4285493755
Semantic Scholar
bfba158f1ad825fe9e5bcc696ce7e71fd8008221
Cited by
200
References
62
Citations by year
2026: 56 · 2025: 64 · 2024: 43 · 2023: 32 · 2022: 5
FWCI
15.07
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • University of Southern Denmark Research Portal (University of Southern Denmark) (submittedVersion, cc-by)
Funders
  • National Research Foundation
  • Agency for Science, Technology and Research (202D800029)
  • National Research Foundation Singapore (NRF-NRFF2017-03)
  • Medical Research Council
  • National Medical Research Council (OFIRG20nov-0085)
Topics
  • Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
  • Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
Keywords
triple-negative breast cancer, tumor hypoxia, HIF-1α, HDAC1, polycomb repressive complex 2, epigenetic suppression, T cell exhaustion, natural killer cells, anti-PD-1 resistance, immunotherapy resistance, tumor microenvironment, immune evasion
MeSH terms
Animals, Hypoxia, Humans, Immunotherapy, Epigenesis, Genetic, Cell Line, Tumor, Mice, Tumor Microenvironment, Triple Negative Breast Neoplasms

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 2026-10-10.

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

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