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Cited by 496Telomeres, Telomerase, and SenescenceImmune cells in cancerCancer Immunotherapy and Biomarkers

Stromal senescence establishes an immunosuppressive microenvironment that drives tumorigenesis

Open access
Ruhland, Megan K., Loza, Andrew J., Capietto, Aude-Helene, Luo, Xianmin, Knolhoff, Brett L., Flanagan, Kevin C., Belt, Brian A., Alspach, Elise +8 more·2016·Nature Communications
DOI 10.1038/ncomms11762PMC PMC4899869
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Abstract

Age is a significant risk factor for the development of cancer. However, the mechanisms that drive age-related increases in cancer remain poorly understood. To determine if senescent stromal cells influence tumorigenesis, we develop a mouse model that mimics the aged skin microenvironment. Using this model, here we find that senescent stromal cells are sufficient to drive localized increases in suppressive myeloid cells that contributed to tumour promotion. Further, we find that the stromal-derived senescence-associated secretory phenotype factor interleukin-6 orchestrates both increases in suppressive myeloid cells and their ability to inhibit anti-tumour T-cell responses. Significantly, in aged, cancer-free individuals, we find similar increases in immune cells that also localize near senescent stromal cells. This work provides evidence that the accumulation of senescent stromal cells is sufficient to establish a tumour-permissive, chronic inflammatory microenvironment that can shelter incipient tumour cells, thus allowing them to proliferate and progress unabated by the immune system.

AI snapshot

Written from the paper’s full text.

TL;DR

Stromal senescence in a mouse model is sufficient to establish an immunosuppressive microenvironment that recruits suppressive myeloid cells and Tregs, inhibits CD8+ T-cell function, and promotes tumour growth. Stromal-derived IL-6 is a key mediator, and aged human skin shows similar senescent stromal cells with increased immune infiltration.

Key findings

  • In FASST mouse skin, senescent stromal cells increased CD45+ immune cells 3.5-fold versus littermate controls, and immune cells localized significantly closer to GFP+ senescent fibroblasts than chance predicted.
  • Senescent MSF isografts increased CD11b+ myeloid cells 4.3-fold and Gr-1+ myeloid cells 3.1-fold; CD11b+Ly6GHi G-MDSCs increased 2.7-fold, FoxP3+ Tregs increased 2.6-fold, and CD8+IFNγ+ cells decreased 2.2-fold. Senescent MSF conditioned medium increased CD11b+Ly6GHi cells 3.5-fold from naive bone marrow.
  • Co-injection of senescent MSFs increased PDSC5 and MK16-Ras tumour growth 2.3- and 6.9-fold, respectively; in nude mice, senescent and non-senescent MSFs promoted tumour growth equivalently (PDSC5 P=0.24, MK16-Ras P=0.49). IL-6 neutralization reduced PDSC5 and MK16-Ras growth 1.8- and 1.9-fold, and shIL-6 senescent MSFs failed to promote tumour growth.
  • In human skin, older donors aged 63–73 years had a 4.3-fold increase in senescent stromal cells and a 3.5-fold increase in CD45+ cells versus younger donors aged 29–32 years, with increased CD33+, CD15+, and CD14+ cells and p16+IL-6+ stromal cells.

Methods

  • A genetically engineered FASST mouse activated p27Kip1 in mesenchymal cells via Col-Cre-ERT2 and ROSA-lox-stop-lox-p27Kip1 after tamoxifen; analyses used 6-week-old mice 2 weeks post-activation, 24-month-old versus 2-month-old mice, and archived human skin samples.
  • Mouse skin fibroblasts (MSFs) were induced to senesce in vitro with tamoxifen; the study used RNA-seq/GSEA, qRT-PCR, ELISA, flow cytometry, immunohistochemistry/immunofluorescence, conditioned medium, bone marrow myeloid conditioning, T-cell suppression assays, and PMA/ionomycin IFNγ stimulation.
  • Subcutaneous isograft experiments co-injected senescent or non-senescent MSFs with PDSC5 or MK16-Ras tumour cells, monitored tumour growth by bioluminescence or calipers, and used neutralizing antibodies against IL-6, Ly6G, and CD8; spatial localization was analysed by nearest-neighbour Monte Carlo sampling.

Limitations

  • The paper states that not all senescence inducers may produce identical immune changes; p16-induced senescence may not lead to SASP expression, and future studies are needed in spontaneous tumour models rather than the isograft model.
  • The paper notes that its nude-mouse results differed from prior published work, possibly due to differences in SASP factors expressed and responding tumour cell types.
  • Evident from the text: human data are retrospective, de-identified archived specimens and correlative, with no causal manipulation in humans; the cell lines were regularly tested for mycoplasma but were not authenticated.

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

Integrity signals

Registry

Crossref · 10.1038/ncomms11762

Cites

Statistics

Nothing to recompute.

Claims the paper makes

0 of 47 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    The stromal-derived SASP factor interleukin-6 orchestrates the ability of suppressive myeloid cells to inhibit anti-tumour T-cell responses.

    “Further, we find that the stromal-derived senescence-associated secretory phenotype factor interleukin-6 orchestrates both increases in suppressive myeloid cells and their ability to inhibit anti-tumour T-cell responses.”
    Quote in the paperSubstantial claim
  • Findingown data

    In the absence of existing tumour cells, senescent stromal cells are sufficient to create an immunosuppressed environment.

    “Upon inducing senescence in the mesenchymal compartment, we find that in the absence of existing tumour cells, senescent stromal cells are sufficient to create an immunosuppressed environment, reminiscent of what we find in aging human skin.”
    Quote in the paperSubstantial claim
  • Findingown data

    Senescent stromal cells are sufficient to drive localized increases in suppressive myeloid cells.

    “Using this model, here we find that senescent stromal cells are sufficient to drive localized increases in suppressive myeloid cells that contributed to tumour promotion.”
    Quote in the paperSubstantial claim
  • Findingown data

    GSEA revealed specific enrichment of factors in the JAK-STAT pathway known to modulate immune cells in senescent MSFs.

    “Gene set enrichment analysis (GSEA) revealed a specific enrichment of factors in the JAK-STAT pathway known to modulate immune cells (P<10−3, GSEA)2425.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [24] Shimozaki, K., Nakajima, K., Hirano, T. & Nagata, S. Involvement of STAT3 in the granulocyte colony-stimulating factor-induced differentiation of myeloid cells. J. Biol. Chem. 272, 25184–25189 (1997). source

      Not checked in the free run.

    • cites [25] Ilaria, R. L. Jr., Hawley, R. G. & Van Etten, R. A. Dominant negative mutants implicate STAT5 in myeloid cell proliferation and neutrophil differentiation. Blood 93, 4154–4166 (1999). source

      Not checked in the free run.

  • Findingown data

    Human skin shows age-dependent increases in stromal p27Kip1 expression.

    “To first verify the relevance of p27Kip1 in age-related senescence, we stained human skin samples and found age-dependent increases in stromal p27Kip1 expression (Supplementary Fig. 1a,b).”
    Quote in the paperSubstantial claim
  • May combine two claims. 34
  • Classified as probably not a substantial claim. 33
  • The quote only partly states this claim. 1

  • May combine two claims.

    Findingown data

    FASST mice show a significant increase in the number of senescent cells within the skin stroma compared with littermate controls, as well as within the mammary gland, lung, and bone.

    “We observed a significant increase in the number of senescent cells within the skin stroma of FASST mice compared with littermate controls (Fig. 1a,b) as well as within the mammary gland, lung (data not shown) and bone20, reflecting the mesenchymal nature of the collagen promoter used in our system.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [20] Luo, X. et al. Stromal-Initiated changes in the bone promote metastatic niche development. Cell Rep. 14, 82–92 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Findingown data

    Cells positive for CD15 were increased in the skin of older individuals, as were CD14+ cells.

    “Further staining indicated that cells positive for CD15 were also increased in the skin of older individuals, as were CD14+ cells (Fig. 10c–f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Skin from 24-month-old mice revealed a significant increase in Ly6G+ cells coincident with increases in SA-βgal positive cells within the stromal compartment.

    “Analysis of skin from 24-month-old mice also revealed a significant increase in Ly6G+ cells (Supplementary Fig. 5a,b) coincident with increases in SA-βgal positive cells within the stromal compartment (Supplementary Fig. 5c,d)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    p16+IL-6+ cells were detected in the older (age 63-73) cohort of donors and there were significant increases in IL-6 within the stromal compartment.

    “We detected p16+IL-6+ cells in the older (age 63–73) cohort of donors as well as significant increases in IL-6 within the stromal compartment (Fig. 9e,f).”
    Quote in the paperSubstantial claim

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

    Findingown data

    Leading-edge analysis suggested multiple genes contributed to the core enrichment of the JAK-STAT pathway.

    “Leading-edge analysis suggested multiple genes contributed to the core enrichment of the pathway (Supplementary Fig. 2a).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    TAM treatment of MSFs induced a senescent and activated stromal phenotype, indicated by transgene expression, growth arrest, increased SA-βgal+ cells, increased IL-6, and elevated FAP.

    “As expected, TAM treatment led to the induction of transgene expression (GFP+), growth arrest, increases in SA-βgal+ cells, increased expression of the canonical SASP-factor IL-6, and elevated expression of fibroblast activation protein (FAP), indicating they are activated stromal cells (Fig. 2a–e and Supplementary Fig. 3a–c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Lack of growth promotion corresponded to the degree of IL-6 knockdown in the individual cell lines, with shIL6 #1 showing the greatest degree of IL-6 knockdown and the greatest reduction of tumour growth promotion compared with the control shRFP line.

    “This lack of growth promotion corresponded to the degree of IL-6 knockdown in the individual cell lines with shIL6 #1 showing the greatest degree of IL-6 knockdown (Fig. 8e) and the greatest reduction of tumour growth promotion compared with the control shRFP line (Fig. 8f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    No significant changes were detected for natural killer (NK) cells, CD4+ or CD8+ T-cell subsets, or CD4/CD8 T-cell ratio.

    “However, no significant changes were detected for natural killer (NK) cells, CD4+ or CD8+ T-cell subsets, or CD4/CD8 T-cell ratio (Supplementary Fig. 5e–h).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Control shRFP-expressing MSFs induced to senesce stained positive for SA-βgal and expressed increased levels of IL-6.

    “When control shRFP-expressing MSFs were induced to senesce, they stained positive for SA-βgal, expressed increased levels of IL-6, (Fig. 8a–e)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    IL-6 is the most highly upregulated SASP factor in MSF gene expression analysis and in vivo isograft gene expression profile.

    “including IL-6 which is the most highly upregulated factor in both our MSF gene expression analysis and in vivo in the isograft gene expression profile (Figs 1c, 3i and Supplementary Fig. 2b).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    No significant changes were detected in the percentages of macrophages (CD11b+MHCII+F4/80+) and dendritic cells (CD11b+MHCII+F4/80-CD11c+).

    “Using CD11b+MHCII+F4/80+ as a marker for macrophages and CD11b+MHCII+F4/80-CD11c+ to identify dendritic cells, we detected no significant changes in the percentages of these myeloid cells (data not shown).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    In non-senescent microenvironments, depletion of Ly6G+ cells had no effect on tumour cell growth while co-depletion of CTLs led to significant increases in tumour cell growth when compared with control antibody.

    “As expected, we found that in non-senescent microenvironments, depletion of Ly6G+ cells had no effect on tumour cell growth while co-depletion of CTLs led to significant increases in tumour cell growth when compared with control antibody (Fig. 6e).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The αCD8 neutralizing antibody efficiently depletes CTLs.

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

    Findingown data

    The increase in CD11b+ myeloid cells with senescent MSFs is consistent with increases seen in the FASST mouse skin.

    “This finding is consistent with increases seen in the FASST mouse skin (Fig. 1h).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    These CD11b+ cells expressed low levels of MHCII and high levels of Ly6G (CD11b+Ly6GHi).

    “Further, these CD11b+ cells expressed low levels of MHCII and high levels of Ly6G (CD11b+Ly6GHi) (Fig. 3a,b).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Presence of control senescent shRFP-expressing MSFs led to increases in G-MDSCs while shIL6-expressing senescent MSFs failed to enrich for this population even in the presence of tumour cells.

    “Importantly, when the inflammatory infiltrate was profiled, the presence of control senescent shRFP-expressing MSFs led to increases in G-MDSCs while shIL6-expressing senescent MSFs failed to enrich for this population (Fig. 8g) even in the presence of tumour cells (Fig. 8h).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Senescent MSFs dramatically increased the number of inflammatory cells and extracellular matrix deposition.

    “In contrast, senescent MSFs dramatically increased the number of inflammatory cells and extracellular matrix deposition (Fig. 2f).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CD45+ immune cells in FASST skin localize preferentially near senescent fibroblasts, with observed median distance significantly smaller than chance.

    “In addition, the increased CD45+ cells appeared to preferentially localize around senescent fibroblasts.”
    • Also stated in: § Senescent stromal cells drive increased inflammation · plain
      “The observed median distance from each immune cell to the nearest senescent fibroblast was found to be significantly smaller than chance would predict in all samples (values>0) (Fig. 1g).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    IL-6 is elevated with age and coincides with increases in both circulating immunosuppressive myeloid cells and cancer incidence.

    “IL-6 is elevated with age and coincides with increases in both circulating immunosuppressive myeloid cells and cancer incidence26.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [2] Ershler, W. B. & Keller, E. T. Age-associated increased interleukin-6 gene expression, late-life diseases, and frailty. Annu. Rev. Med. 51, 245–270 (2000). source

      Not checked in the free run.

    • cites [6] Verschoor, C. P. et al. Blood CD33(+)HLA-DR(−) myeloid-derived suppressor cells are increased with age and a history of cancer. J. Leukoc. Biol. 93, 633–637 (2013). source

      Not checked in the free run.

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

    Premisecitation

    In some tumour models, senescent neoplastic cells can stimulate immune-mediated tumour cell clearance, so senescence functions as a tumour-suppressive mechanism.

    “Indeed, in some tumour models, senescent neoplastic cells can stimulate immune-mediated tumour cell clearance and thus, in this context, senescence functions as a potent tumour-suppressive mechanism13.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13] Xue, W. et al. Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature 445, 656–660 (2007). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Accumulation of senescent stromal cells is sufficient to establish a tumour-permissive, chronic inflammatory microenvironment that can shelter incipient tumour cells, allowing them to proliferate and progress unabated by the immune system.

    “This work provides evidence that the accumulation of senescent stromal cells is sufficient to establish a tumour-permissive, chronic inflammatory microenvironment that can shelter incipient tumour cells, thus allowing them to proliferate and progress unabated by the immune system.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Senescent cells accumulate with age.

    “It remains unclear what drives these increases, but one contributing factor may be the accumulation of senescent cells that is known to occur with age789.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [7] Dimri, G. P. et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl Acad. Sci. USA 92, 9363–9367 (1995). source

      Not checked in the free run.

    • cites [8] Wang, C. et al. DNA damage response and cellular senescence in tissues of aging mice. Aging Cell 8, 311–323 (2009). source

      Not checked in the free run.

    • cites [9] Jeyapalan, J. C., Ferreira, M., Sedivy, J. M. & Herbig, U. Accumulation of senescent cells in mitotic tissue of aging primates. Mech. Ageing Dev. 128, 36–44 (2007). source

      Not checked in the free run.

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

    Premisecitation

    Senescent cells express p16INK4A, senescence-associated β-galactosidase, and an altered expression profile known as the senescence-associated secretory phenotype.

    “In addition, senescent cells express the cell cycle inhibitor p16INK4A (p16), senescence-associated β-galatosidase (SA-βgal), and an altered expression profile known as the senescence-associated secretory phenotype (SASP)11.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [11] Coppe, J. P., Desprez, P. Y., Krtolica, A. & Campisi, J. The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu. Rev. Pathol. 5, 99–118 (2010). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    Age is a significant risk factor for the development of cancer.

    “Age is a significant risk factor for the development of cancer.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [1] DePinho, R. A. The age of cancer. Nature 408, 248–254 (2000). source

      Not checked in the free run.

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

    Premisecitation

    Among SASP cytokines, interleukin-6 is considered a canonical inflammatory factor.

    “Among the SASP cytokines, interleukin-6 (IL-6) is considered a canonical inflammatory factor12.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [12] Naugler, W. E. & Karin, M. The wolf in sheep's clothing: the role of interleukin-6 in immunity, inflammation and cancer. Trends Mol. Med. 14, 109–119 (2008). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Senescence plays a paradoxical role in tumorigenesis, being both tumour-promoting and tumour-suppressive depending on the cell in which senescence occurs.

    “Senescence plays a paradoxical role in tumorigenesis, being both tumour-promoting and tumour-suppressive depending on the cell in which senescence occurs.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    p16INK4a is a well-characterized senescence marker used extensively in human tissue.

    “we turned to the well-characterized senescence maker p16INK4a (p16), which is used extensively in human tissue2336.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [2336]

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Increases in G-MDSCs and Treg cells along with decreases in CD8+ T-cell function indicate that senescent stromal cells were sufficient to create an immunosuppressive microenvironment.

    “Collectively, increases in G-MDSCs and Treg cells along with decreases in CD8+ T-cell function indicate that senescent stromal cells were sufficient to create an immunosuppressive microenvironment.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Senescence plays an important role in biological processes ranging from patterning in developing embryos and wound healing, to tumour prevention and tumour promotion.

    “Senescence plays an important role in biological processes ranging from patterning in developing embryos and wound healing, to tumour prevention and tumour promotion38394041424344.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites 38394041424344

      Not checked in the free run.

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

    Premisecitation

    CD15 and CD14 can be used to identify granulocytic and monocytic cells, respectively.

    “The cell surface proteins CD15 and CD14 can be used to further identify granulocytic and monocytic cells, respectively37.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [37] Greten, T. F., Manns, M. P. & Korangy, F. Myeloid derived suppressor cells in human diseases. Int. Immunopharmacol. 11, 802–807 (2011). source

      Not checked in the free run.

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

    Premisecitation

    Senescent cells increase in many tissues including the skin of humans as they age.

    “Senescent cells increase in many tissues including the skin of humans as they age7.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [7] Dimri, G. P. et al. A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc. Natl Acad. Sci. USA 92, 9363–9367 (1995). source

      Not checked in the free run.

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

    Premisecitation

    CD11b+Ly6GHi and CD11b+Ly6CHi myeloid cells are phenotypically consistent with granulocytic and monocytic MDSCs and are often associated with immunosuppression.

    “CD11b+Ly6GHi and CD11b+Ly6CHi myeloid cells are phenotypically consistent with granulocytic and monocytic MDSCs and are often associated with immunosuppression2728.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [2728]

      Not checked in the free run.

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

    Premisecitation

    IFNγ promotes increased tumour immunogenicity and is often used to assess T-cell activation.

    “a cytokine which promotes increased tumour immunogenicity and often used to assess T-cell activation31”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [31] Dunn, G. P., Koebel, C. M. & Schreiber, R. D. Interferons, immunity and cancer immunoediting. Nat. Rev. Immunol. 6, 836–848 (2006). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Within aging human skin, senescent stromal cells express IL-6 and may affect local inflammation.

    “These finding demonstrate that within aging human skin, senescent stromal cells express IL-6 and may affect local inflammation.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    These myeloid cell subsets have been implicated in the induction of other immunosuppressive cell types including FOXP3+ T regulatory cells (Treg).

    “In addition, these myeloid cell subsets have been implicated in the induction of other immunosuppressive cell types including FOXP3+ T regulatory cells (Treg) (refs 28, 29).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites refs 28, 29

      Not checked in the free run.

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

    Study design and conductown data

    Upregulation of factors was validated using quantitative reverse transcription PCR.

    “Upregulation of factors was validated using quantitative reverse transcription PCR (qRT-PCR) (Supplementary Fig. 2b).”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Suppressive myeloid cells increase with age in the circulation of humans and are elevated within the spleens of older mice.

    “recent evidence shows that suppressive myeloid cells increase with age in the circulation of humans and are elevated within the spleens of older mice63435.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites 63435

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    STAT3 can drive SASP expression as well as immunosuppressive inflammation.

    “the transcription factor STAT3 that can drive SASP expression as well as immunosuppressive inflammation30”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] Toso, A. et al. Enhancing chemotherapy efficacy in Pten-deficient prostate tumors by activating the senescence-associated antitumor immunity. Cell Rep. 9, 75–89 (2014). source

      Not checked in the free run.

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

    Study design and conductnone given

    Mice used in the study were subject to speed congenics and bred to an FVB background.

    “Finally, mice were subject to speed congenics and bred to an FVB background.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Campisi and colleagues have shown that unlike chronologically, ARF, or Ras-induced senescence, p16-induced senescence does not lead to SASP expression.

    “Campisi and colleagues have shown that unlike chronologically, ARF, or Ras-induced senescence, p16-induced senescence does not lead to SASP expression48.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [48] Coppe, J. P. et al. Tumor suppressor and aging biomarker p16(INK4a) induces cellular senescence without the associated inflammatory secretory phenotype. J. Biol. Chem. 286, 36396–36403 (2011). source

      Not checked in the free run.

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

    Premisecitation

    The mechanisms responsible for the paradoxical role that senescence plays in tumorigenesis have remained obscure but are likely to lie in the senescent secretory profile that is defined by the cell of origin.

    “The mechanisms responsible for the paradoxical role that senescence plays in tumorigenesis have remained obscure but are likely to lie in the senescent secretory profile that is defined by the cell of origin11.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [11] Coppe, J. P., Desprez, P. Y., Krtolica, A. & Campisi, J. The senescence-associated secretory phenotype: the dark side of tumor suppression. Annu. Rev. Pathol. 5, 99–118 (2010). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    In spatial analysis, regions other than the interstitial stroma (outside tissue, epithelial layer, hair follicles, muscle tissue) were masked before processing.

    “Before processing, regions of the image other than the interstitial stroma were masked (outside the tissue, the epithelial layer, hair follicles, muscle tissue) were masked.”
    Quote in the paperSubstantial claim

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

    Study design and conductown data

    Several transformations (log, square root and Box-Cox) on the tumour size measurements were analysed and all gave similar conclusions on the senescence effect.

    “Note that several transformations (log, square root and Box-Cox) on the tumour size measurements were analysed and all gave similar conclusions on the senescence effect.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    The Box-Cox transformation (with the parameter estimated as 0.6747 using the R function 'boxcox') resulted in the best goodness-of-fit and was reported.

    “The Box-Cox transformation (with the parameter estimated as 0.6747 using the R function ‘boxcox') resulted in the best goodness-of-fit and was reported.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Clearance of senescent cells may be tissue-specific, cell type-specific and/or specific to the mutational profile of the transformed cells in question.

    “However, these cells were not cleared, suggesting that clearance of senescent cells may be tissue-specific, cell type-specific and/or specific to the mutational profile of the transformed cells in question.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    In a PTEN-null prostate cancer model, robust induction of senescence was observed within tumour cells throughout the tissue.

    “In a PTEN-null, prostate cancer model, again robust induction of senescence was observed within tumour cells throughout the tissue30.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [30] Toso, A. et al. Enhancing chemotherapy efficacy in Pten-deficient prostate tumors by activating the senescence-associated antitumor immunity. Cell Rep. 9, 75–89 (2014). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    A recent study demonstrated that senescence represents an important tumour-promoting mechanism in aging animals.

    “a recent study demonstrated that senescence represents an important tumour-promoting mechanism in aging animals.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [10] Baker, D. J. et al. Naturally occurring p16-positive cells shorten healthy lifespan. Nature 530, 184–189 (2016). source

      Not checked in the free run.

  • Classified as probably not a substantial claim. The quote only partly states this claim.

    Study design and conductnone given

    For quantification of CD31, Ly6G, and human IL-6/p16 immunofluorescence staining, the stromal region was selected for mean fluorescence intensity quantification, excluding the epithelium from the analysis.

    “Quantification of staining was performed using Fiji software to analyse mean fluorescence intensity (MFI) per image for CD31. Before calculating the MFI, the freehand selection tool was used to select only the stromal region for MFI quantification thus excluding the epithelium from the analysis.”
    • Also stated in: § Immunohistochemistry and immunofluorescence staining · plain
      “Quantification of Ly6G staining was performed using a similar stromal selection technique.”
    • Also stated in: § Immunohistochemistry and immunofluorescence staining · plain
      “Quantification of staining was performed using Fiji software to analyse MFI per image. Before calculating the MFI, the freehand selection tool was used to select only the stromal region for MFI quantification thus excluding the epithelium from the analysis.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Xue et al. demonstrated that within the liver, senescent carcinoma cells elicited immune-mediated clearance that restrained tumour growth.

    “Xue et al. demonstrated that within the liver, senescent carcinoma cells elicited immune-mediated clearance that restrained tumour growth13.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [13] Xue, W. et al. Senescence and tumour clearance is triggered by p53 restoration in murine liver carcinomas. Nature 445, 656–660 (2007). source

      Not checked in the free run.

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

    Premisecitation

    Secretion of inflammatory cytokines can elicit immune-mediated clearance of senescent tumour cells.

    “In addition to directly limiting replicative capacity, in this setting the secretion of inflammatory cytokines can elicit immune-mediated clearance of senescent tumour cells46.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [46] Iannello, A., Thompson, T. W., Ardolino, M., Lowe, S. W. & Raulet, D. H. p53-dependent chemokine production by senescent tumor cells supports NKG2D-dependent tumor elimination by natural killer cells. J. Exp. Med. 210, 2057–2069 (2013). source

      Not checked in the free run.

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

    Study design and conductreasoning

    In RNA-seq analysis, log2 fold changes were computed after applying a floor of 1 c.p.m. to ensure that genes with very low overall expression but large relative fold changes did not affect the analysis.

    “Log2 fold changes were computed after applying a floor of 1 c.p.m. to ensure that genes with very low over all expression, but large relative fold changes did not affect the analysis.”
    Quote in the paperNot a substantial claim

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

    Study design and conductreasoning

    In spatial analysis, the median distance from immune cells to the nearest senescent cell was chosen over the mean to make the distance statistically less susceptible to outliers.

    “The median was chosen over the mean to make this distance statistically less susceptible to outliers.”
    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

    All cell lines used in the study were regularly tested for mycoplasma but were not authenticated.

    “All cell lines were regularly tested for mycoplasma but were not authenticated.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Senescent stromal cells create an immunosuppressive environment characterized by the accumulation of MDSCs that limit CD8+ T-cell responses and allow tumour cell growth.

    “Indeed we demonstrate that senescent stromal cells create an immunosuppressive environment characterized by the accumulation of MDSCs that limit CD8+ T-cell responses and allow tumour cell growth.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    Stromal-derived SASP-factor IL-6 establishes myeloid-driven immunosuppression where CD8+ T cells are inhibited, resulting in unrestrained tumour growth.

    “In our work, we found that the stromal-derived SASP-factor IL-6 establishes myeloid-driven immunosuppression where CD8+ T cells were inhibited, resulting in unrestrained tumour growth.”
    Quote in the paperSubstantial claim

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

    Interpretationown data

    Our work provides a possible mechanism to describe these findings.

    “Our work provides a possible mechanism to describe these findings.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    The T-cell suppression experiments were performed in triplicate.

    “Experiments were performed in triplicate.”
    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 conductown data

    Growth curve data are representative of two independent experiments and are presented as mean ± s.e.m.

    “Data shown are representative of two independent experiments and presented as mean ± s.e.m.”
    Quote in the paperNot a substantial claim
57 of 57

  • The paper relies on this without support.

    Nude mice lack T cells and therefore the equivalent tumour promotion by senescent and non-senescent MSFs in nude mice reflects the absence of an adaptive immune system.

    “we co-injected tumour cells and MSFs into nude mice”
    “the tumour promoting activities of senescent and non-senescent MSFs were equivalent”

  • The paper relies on this without support.

    The αCD8 neutralizing antibody efficiently depletes CTLs.

    “using a neutralizing antibody against CD8α, we tested the consequences of a cytotoxic T lymphocyte (CTL) response on tumour growth”
    “simultaneous depletion of CD8+ and Ly6G+ cells”

  • The paper relies on this without support.

    The factors in senescent MSF-conditioned medium that drive myeloid changes are SASP factors.

    “we generated conditioned medium (CM) from non-senescent or senescent MSFs and placed it on naive bone marrow cells for 96 h”
    “CM from senescent MSFs led to a 3.5-fold increase in CD11b+Ly6GHi cells”

  • The paper relies on this without support.

    The αLy6G antibody specifically depletes G-MDSCs without affecting other cell types.

    “we used a Ly6G neutralizing antibody (αLy6G, clone 1A8) to deplete G-MDSCs”
    “our depletion approach efficiently eliminated greater than 85% of these cells”

  • The paper relies on this without support.

    The accumulation of suppressive myeloid cells within senescent tissues of older individuals can precede the appearance of a tumour.

    “However, whether the accumulation of suppressive myeloid cells within senescent tissues of older individuals can precede the appearance of a tumour remains an open question.”

  • The paper relies on this without support.

    CD45 staining identifies leukocytes in human skin.

    “We co-stained normal human skin of varying age with antibodies against p16 (red) and CD45 (green) to identify senescent cells and leukocytes, respectively.”
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?

  • Senescent cells are present in tumor microenvironments across cancers and can influence immune infiltration and therapy outcomes.

    “Senescent cells are present in tumor microenvironments (TMEs) across cancers and can influence immune infiltration and therapy outcomes.57,58,59”

    From Cellular senescence in human liver under normal aging and cancer (2026) · cites this paper as [59]

    The Abstract and Results report that senescent stromal cells alter suppressive immune-cell populations and inhibit anti-tumour T-cell responses in a mouse model. The paper also discusses senescence in several tumour settings, but it does not establish that senescent cells are present in tumour microenvironments across cancers or show an effect on therapy outcomes. Page numbers are not available in our copy. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • IL-6-secreting senescent cells residing in the tumour stroma recruit myeloid suppressor cells (expressing CD11b and Gr-1) and reduce antitumour T cell immune surveillance.

    “IL-6- secreting senescent cells residing in the tumour stroma were shown to recruit myeloid suppressor cells (expressing CD11b and Gr-1) and reduce antitumour Tcell immune surveillance 175 .”

    From Senescence and cancer — role and therapeutic opportunities (2022) · cites this paper as [175]

  • This occurs via recruitment of myeloid-derived suppressor cells chemoattracted by factors such as IL-6, IL-8, and CCL2 secreted by senescent cells in murine models of hepatocellular, prostate, and skin cancers.

    “There is convergent evidence that this occurs via the recruitment of myeloid-derived suppressor cells (MDSCs) chemoattracted by factors (e.g., IL-6, IL-8, and CCL2) secreted by senescent cells in murine models of HCC, 193 prostate, 194 and skin cancers.195”

    From Meta-hallmarks of aging and cancer (2023) · cites this paper as [195]

  • The SASP can create a pro-tumorigenic environment.

    “On the other hand, the SASP can create a pro-tumorigenic environment (Coppe et al., 2010; Krtolica et al., 2001; Ruhland et al., 2016).”

    From CDK4/6 Inhibitors: The Mechanism of Action May Not Be as Simple as Once Thought (2018) · cites this paper as Ruhland et al., 2016

  • This leads to the inhibition of anti-tumour T-cell responses and enhanced tumour growth.

    “Furthermore, it was shown that this leads to the inhibition of anti-tumour T-cell responses and enhanced tumour growth.77”

    From Paracrine roles of cellular senescence in promoting tumourigenesis (2018) · cites this paper as [77]

The citation lists haven’t been fetched yet

References 54 · Cited by 496

Authors
Ruhland, M. K., Loza, A. J., Capietto, A.-H., Luo, X., Knolhoff, B. L., Flanagan, K. C., Belt, B. A., Alspach, E., Leahy, K., Luo, J., Schaffer, A., Edwards, J. R., Longmore, G., Faccio, R., DeNardo, D. G., & Stewart, S. A.
Affiliations
Washington University in St. Louis (US)
Corresponding author
Stewart, Sheila A.
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2016-06-08
Volume
7
Issue
1
Pages
11762-11762
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/ncomms11762
PMC
PMC4899869
OpenAlex
W2411937682
Cited by
496
References
54
Citations by year
2026: 60 · 2025: 72 · 2024: 79 · 2023: 60 · 2022: 56 · 2021: 55 · 2020: 37 · 2019: 28 · 2018: 29 · 2017: 15 · 2016: 4 · 2012: 1
FWCI
20.40
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Commons@Becker (Washington University School of Medicine) (submittedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
  • American Cancer Society
  • Susan G. Komen for the Cure
  • Washington University in St. Louis (P30 CA091842)
  • Foundation for Barnes-Jewish Hospital (P30 CA091842)
  • American Society for Bone and Mineral Research
  • Alvin J. Siteman Cancer Center (P30CA091842, CA091842)
  • National Institutes of Health (NIH P50, CA091842, CA143868, CA94056)
  • National Cancer Institute (U54CA143868, R01CA151518, P50 CA94056, P50CA094056, CA-94056, CA091842, R01CA177670, R01CA203890, R01CA196205, T32CA113275, P30CA091842)
  • National Institute of General Medical Sciences (R01GM108811, T32GM007200)
  • National Institute of Arthritis and Musculoskeletal and Skin Diseases (P30AR057235)
  • National Institute of Biomedical Imaging and Bioengineering (T32EB018266)
Topics
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
  • Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
Keywords
carcinogenesis, immunosuppressive tumor microenvironment, interleukin-6, senescence-associated secretory phenotype, aging and cancer, chronic inflammation, aged skin, tumor promotion
MeSH terms
Myeloid-Derived Suppressor Cells, Adult, Animals, Antigens, Ly, Cell Line, Fibroblasts, Humans, Immunologic Surveillance, Immunosuppression Therapy, Inflammation, Middle Aged, Skin, Interleukin-6, Cellular Senescence, Stromal Cells, CD11b Antigen, Cell Proliferation, T-Lymphocytes, Regulatory, Mice, Tumor Microenvironment, Carcinogenesis

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Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-07.

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