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Cited by 988Neutrophil, Myeloperoxidase and Oxidative MechanismsImmune cells in cancerCancer Immunotherapy and Biomarkers

Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps

Open access
Park, Juwon, Wysocki, Robert W., Amoozgar, Zohreh, Maiorino, Laura, Fein, Miriam R., Jorns, Julie, Schott, Anne F., Kinugasa-Katayama, Yumi +11 more·2016·Science Translational Medicine
DOI 10.1126/scitranslmed.aag1711PMC PMC5550900
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Abstract

Treatment with DNase I–coated nanoparticles prevents metastasis by targeting neutrophil extracellular traps induced by cancer cells in a mouse model.

AI snapshot

Written from the paper’s full text.

TL;DR

Metastatic breast cancer cells induce neutrophils to form NETs in the absence of infection, and these NETs promote cancer cell invasion and lung metastasis. Digesting NETs with DNase I-coated nanoparticles reduced metastatic burden in mice, suggesting NETs as a therapeutic target.

Key findings

  • In mice, metastatic 4T1 cells induced NET-like extracellular DNA structures in the lungs, with neutrophil elastase activity co-localizing with extracellular DNA in 14 of 17 imaged fields after 4T1 injection but not after non-metastatic 4T07 injection (p=0.0008).
  • In human breast cancer samples, NETs were detected in 16 of 20 primary tumors and 13 of 19 metastatic lung lesions; the highest numbers were in triple-negative tumors (6 of 6 primary tumors and 7 of 7 metastases), with absent or very low numbers in luminal cancers.
  • NETs stimulated breast cancer cell invasion and migration in vitro; DNase I digestion blocked these effects. Cancer cell-induced NET formation required G-CSF, NADPH oxidase, PAD4, cathepsin G, and neutrophil elastase, and occurred through a lytic process.
  • DNase I-coated nanoparticles significantly reduced experimental lung metastasis: 3 of 9 treated mice had no detectable metastasis versus all 10 control mice, with reduced metastatic foci number (p=0.0003) and average foci size (p=0.01). In spontaneous metastasis, 3 of 6 treated mice had no detectable metastases versus all control mice having micrometastases, with no effect on primary tumor growth.

Methods

  • In vitro transwell invasion/migration and NET-formation assays used murine 4T1, 4T07, and C3(1)-Tag breast cancer cells and human BT-549 cells with mouse or human neutrophils, plus inhibitors, blocking antibodies, DNase I, conditioned medium, immunofluorescence, and electron microscopy.
  • In vivo studies used confocal intravital lung imaging (CILI) in LysM-EGFP mice, experimental and spontaneous 4T1 metastasis models in BALB/c mice, and daily intraperitoneal DNase I-coated nanoparticles (75 U/mouse); clinical breast cancer samples were analyzed by immunofluorescence.
  • Statistical analyses used GraphPad Prism with two-sided t-tests or one-way ANOVA at alpha 0.05; some assessments were blinded, and sample sizes are in figure legends.

Limitations

  • Stated: the p47phox−/− strain is on the C57BL/6 background, precluding in vivo experiments with BALB/c-derived 4T1 cells.
  • Stated: long-term PAD4 inhibition could not be tested for metastasis because commercially available PAD4 inhibitors have serum half-lives of ∼15 min–4 h, so DNase I-coated nanoparticles could not be compared with PAD4 inhibition as strategies.
  • Stated/evident: the clinical sample panel was small, and the paper notes that assays are needed to identify patients who may benefit from NET-targeted treatments; it also states that whether NETs participate in migration and invasion in the primary tumor remains a future direction.

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

Integrity signals

Registry

Crossref · 10.1126/scitranslmed.aag1711

Cites

50 of 50 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 27 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    Treatment with NET-digesting, DNase I-coated nanoparticles markedly reduces lung metastases in mice.

    “Treatment with NET-digesting, DNase I-coated nanoparticles markedly reduced lung metastases in mice.”
    • Also stated in: § DNase I treatment prevents lung metastasis in mice · plain
      “Daily intraperitoneal injection of DNase I-coated nanoparticles significantly (p=0.002) reduced metastatic burden after intravenous injection of 4T1 cells (Figs. 6C, D).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: DNase I-coated nanoparticles reduce metastasis specifically by digesting NETs.
  • Findingreasoning

    CXCL1/2 knockdown does not increase primary tumor growth in T cell-deficient nude mice.

    “Consistent with this idea, CXCL1/2 knockdown did not increase primary tumor growth in T cell-deficient, nude mice (Fig. 1E).”
    Quote in the paperSubstantial claim
  • Findingown data

    The formation of NETs stimulates the invasion and migration of breast cancer cells in vitro.

    “The formation of NETs stimulated the invasion and migration of breast cancer cells in vitro.”
    • Also stated in: § NETs promote cancer cell migration and invasion · plain
      “Co-culturing with neutrophils increased the invasion of 4T1 cells, but had little effect on the invasion of 4T07 cells (Fig. 3C).”
    Quote in the paperSubstantial claim
  • Findingown data

    CXCL1/2 knockdown significantly reduces spontaneous metastasis in nude mice (p=0.03).

    “However, CXCL1/2 knockdown did significantly (p=0.03) reduce spontaneous metastasis in nude mice (Fig. 1F).”
    Quote in the paperSubstantial claim
  • Findingown data

    Neutrophil elastase activity co-localizes with extracellular DNA in 14 out of 17 imaged fields after injection of 4T1 cancer cells but not after injection of 4T07 cells (p=0.0008).

    “Neutrophil elastase activity co-localized with extracellular DNA in 14 out of 17 imaged fields of view after injection of the 4T1 cancer cells but not after injection of 4T07 cells (Figs. 1I, J, p=0.0008).”
    Quote in the paperSubstantial claim
  • May combine two claims. 25
  • Classified as probably not a substantial claim. 19

  • May combine two claims.

    Findingown data

    Inhibiting NET formation or digesting NETs with DNase I blocks NET-stimulated invasion and migration of breast cancer cells.

    “Inhibiting NET formation or digesting NETs with DNase I blocked these processes.”
    • Also stated in: § NETs promote cancer cell migration and invasion · plain
      “The neutrophils' ability to stimulate invasion of 4T1 cells was lost when the DNA of the NETs was digested, whereas the addition of DNase I had no effect on fetal calf serum (FCS)-stimulated invasion (Fig. 3F).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Reduction of CXCL1/2 in 4T1 cells has no effect on macrophage infiltration or cancer cell proliferation in vitro.

    “Reduction of CXCL1 (and the homologous CXCL2) in 4T1 cells with short hairpin RNAs reduced neutrophil infiltration into tumors and increased primary tumor growth, but had no effect on macrophage infiltration or cancer cell proliferation in vitro (fig. S1).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    NETs are detected in 16 out of 20 primary tumors and in 13 out of 19 metastatic lung lesions from breast cancer patients.

    “We detected NETs in 16 out of 20 primary tumors, and in 13 out of 19 metastatic lung lesions (Fig. 2B).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The immunofluorescence staining used on human clinical samples detects NETs specifically.

  • May combine two claims.

    Findingown data

    NETs form in lungs shortly after tail vein injection of 4T1 cells and the amounts of NETs are elevated for days after injection.

    “This analysis confirmed that NETs formed in lungs shortly after tail vein injection of 4T1 cells and that the amounts of NETs were elevated for days after injection (Figs. 1K, 1L, and S2).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    CXCL1 mRNA and protein are higher in 4T1 than in 4T07 cells and in 4T1-derived tumors than in 4T07-derived tumors.

    “we measured its mRNA and protein and found higher amounts in 4T1 than in 4T07 cells and in 4T1-derived tumors than in 4T07-derived tumors (Fig. 1C, and fig. S1).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Co-culturing with neutrophils increases the invasion of 4T1 cells but has little effect on the invasion of 4T07 cells.

    “Co-culturing with neutrophils increased the invasion of 4T1 cells, but had little effect on the invasion of 4T07 cells (Fig. 3C).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Neutrophils co-cultured with metastatic 4T1 cells form extensive NETs, whereas neutrophils co-cultured with non-metastatic 4T07 cells form few NETs.

    “Neutrophils co-cultured in this manner with metastatic 4T1 cells formed extensive NETs, whereas neutrophils similarly co-cultured with non-metastatic 4T07 cells formed few NETs (Figs. 3A, B).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The number of NETs varies between tumors, with the highest numbers in triple-negative tumors and an absence or very low numbers in luminal breast cancers.

    “The number of NETs varied between tumors, with the highest numbers in triple-negative tumors (6 of 6 primary tumors and 7 of 7 metastases had detectable NETs), and an absence or very low numbers in luminal breast cancers (Fig. 2C).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The immunofluorescence staining used on human clinical samples detects NETs specifically.

  • May combine two claims.

    Findingown data

    Inhibition of cathepsin G blocks the neutrophils' ability to promote invasion with no effect on the cancer cells' ability to invade towards FCS.

    “Indeed, inhibition of cathepsin G, a protease associated with NETs, blocked the neutrophils' ability to promote invasion with no effect on the cancer cells' ability to invade towards FCS.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Three out of nine mice treated with DNase I-coated nanoparticles have no detectable metastasis by histology, whereas all 10 mice treated with control nanoparticles have macroscopic or microscopic metastases.

    “Three out of nine mice treated with DNase-I-coated nanoparticles had no detectable metastasis by histology, whereas all 10 mice treated with control nanoparticles had macroscopic or microscopic metastases.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Conditioned medium from cultures induced to form NETs effectively stimulates cancer cell invasion, whereas conditioned medium from cultures where NET formation is inhibited with apocynin does not.

    “CM from cultures induced to form NETs effectively stimulated cancer cell invasion, whereas CM from cultures where NET formation was inhibited with apocynin did not (Fig. 5F).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Conditioned medium loses the ability to stimulate invasion if DNase I is added after collection or upon heat denaturing.

    “In addition, the CM lost its ability to stimulate invasion if DNase I was added after collection or upon heat denaturing (fig. S6).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Despite a significant reduction in the number of NETs using DNase I-coated nanoparticles or Cl-amidine (p=0.02), the number of cells that extravasated into the lung tissue 24 hours after injection is not altered.

    “Despite a significant (p=0.02) reduction in the number of NETs using either approach (fig. S7A), the number of cells that extravasated into the lung tissue 24 hours after injection was not altered (fig. S7B).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    DNase I digestion and heat denaturing abolish the invasion-promoting effect of conditioned medium from NET-forming cultures.

    “We found that conditioned medium from NET-forming cultures can promote invasion, but that both DNase I digestion and heat denaturing can abolish this effect.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    In a spontaneous metastasis model, lung metastatic burden is reduced: all mice treated with control nanoparticles have micrometastases, but 3 out of 6 mice treated with DNase I-coated nanoparticles have no detectable metastases by histology.

    “The DNase I-coated nanoparticles had no effect on primary tumor growth (Fig. 6G), but the lung metastatic burden was reduced: all mice treated with control nanoparticles had micrometastases, but 3 out of 6 mice treated with DNase I-coated nanoparticles had no detectable metastases by histology (Fig. 6H).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Digesting bacterially induced NETs with systemically administered DNase I can reduce experimental metastasis in a mouse model of metastatic lung cancer, but the effects are relatively modest.

    “Digesting bacterially induced NETs with systemically administered DNase I can reduce experimental metastasis in a mouse model of metastatic lung cancer, but the effects are relatively modest (5).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [5] Cools-Lartigue J Spicer J McDonald B Gowing S Chow S Giannias B Bourdeau F Kubes P Ferri L Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis J Clin Invest 123 3446 3458 2013 PMC3726160 23863628 source

      Not checked in the free run.

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

    Study design and conductnone given

    The p47phox-/- strain is on the C57BL/6 background, precluding in vivo experiments with the BALB/c-derived 4T1 cells.

    “Unfortunately, the p47phox-/- strain is on the C57BL/6 background, precluding in vivo experiments with the BALB/c-derived 4T1 cells.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    One type of NET formation requires NADPH oxidase activity and generally results in neutrophil lysis 2-4 hours after activation.

    “One requires NADPH oxidase activity and generally results in neutrophil lysis 2-4 hours after activation (24), whereas another process occurs rapidly (5-60 min), independent of NADPH oxidase-activity, and leaves the neutrophil plasma membrane intact (26).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] Fuchs TA Abed U Goosmann C Hurwitz R Schulze I Wahn V Weinrauch Y Brinkmann V Zychlinsky A Novel cell death program leads to neutrophil extracellular traps J Cell Biol 176 231 241 2007 17210947 PMC2063942 source

      Not checked in the free run.

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

    Premisecitation

    NETs that form intravascularly in response to infection can promote extravasation of disseminated cancer cells.

    “NETs that form intravascularly in response to infection can promote extravasation of disseminated cancer cells (5).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [5] Cools-Lartigue J Spicer J McDonald B Gowing S Chow S Giannias B Bourdeau F Kubes P Ferri L Neutrophil extracellular traps sequester circulating tumor cells and promote metastasis J Clin Invest 123 3446 3458 2013 PMC3726160 23863628 source

      Not checked in the free run.

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

    Premisecitation

    Another type of NET formation occurs rapidly (5-60 min), is independent of NADPH oxidase activity, and leaves the neutrophil plasma membrane intact.

    “One requires NADPH oxidase activity and generally results in neutrophil lysis 2-4 hours after activation (24), whereas another process occurs rapidly (5-60 min), independent of NADPH oxidase-activity, and leaves the neutrophil plasma membrane intact (26).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [26] Pilsczek FH Salina D Poon KK Fahey C Yipp BG Sibley CD Robbins SM Green FH Surette MG Sugai M Bowden MG Hussain M Zhang K Kubes P A novel mechanism of rapid nuclear neutrophil extracellular trap formation in response to Staphylococcus aureus J Immunol 185 7413 7425 2010 21098229 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    G-CSF can prime neutrophils for NET formation and is secreted by 4T1 cells.

    “G-CSF can prime neutrophils for NET formation and is secreted by 4T1 cells (23).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [23] Demers M Krause DS Schatzberg D Martinod K Voorhees JR Fuchs TA Scadden DT Wagner DD Cancers predispose neutrophils to release extracellular DNA traps that contribute to cancer-associated thrombosis Proc Natl Acad Sci U S A 109 13076 13081 2012 22826226 PMC3420209 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Both human and mouse triple-negative breast cancer cells can promote the formation of pro-migratory/invasive NETs.

    “Thus, both human and mouse triple-negative breast cancer cells can promote the formation of pro-migratory/invasive NETs, whereas treatment with NET-digesting DNase I blocks migration and invasion induced by species-matched neutrophils.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    An initial step in the intracellular signaling cascade in neutrophils that results in pathogen-induced NET formation is activation of the phagocytic NADPH oxidase (NOX2) enzyme complex.

    “An initial step in the intracellular signaling cascade in neutrophils that results in pathogen-induced NET formation is activation of the phagocytic NADPH oxidase (NOX2) enzyme complex (24).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [24] Fuchs TA Abed U Goosmann C Hurwitz R Schulze I Wahn V Weinrauch Y Brinkmann V Zychlinsky A Novel cell death program leads to neutrophil extracellular traps J Cell Biol 176 231 241 2007 17210947 PMC2063942 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Metastatic cancer cells activate signaling pathways that include NADPH oxidase and PAD4 in neutrophils, causing the formation of invasion-promoting NETs.

    “Thus, metastatic cancer cells activate signaling pathways that include NADPH oxidase and PAD4 in neutrophils, causing the formation of invasion-promoting NETs.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Cleavage of histones by neutrophil elastase is required for chromatin de-condensation and NET release.

    “Cleavage of histones by neutrophil elastase is required for chromatin de-condensation and NET release (28, 29), and cathepsin G may have a similar role.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [28] Papayannopoulos V Metzler KD Hakkim A Zychlinsky A Neutrophil elastase and myeloperoxidase regulate the formation of neutrophil extracellular traps J Cell Biol 191 677 691 2010 20974816 PMC3003309 source

      Not checked in the free run.

    • cites [29] Metzler KD Goosmann C Lubojemska A Zychlinsky A Papayannopoulos V A myeloperoxidase-containing complex regulates neutrophil elastase release and actin dynamics during NETosis Cell Rep 8 883 896 2014 25066128 PMC4471680 source

      Not checked in the free run.

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

    Premisecitation

    Intravascular NETs can increase local vascular permeability.

    “In addition, intravascular NETs can increase local vascular permeability (15), which would permit cancer cells to extravasate more easily.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [15] Kolaczkowska E Jenne CN Surewaard BG Thanabalasuriar A Lee WY Sanz MJ Mowen K Opdenakker G Kubes P Molecular mechanisms of NET formation and degradation revealed by intravital imaging in the liver vasculature Nat Commun 6 6673 2015 25809117 PMC4389265 source

      Not checked in the free run.

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

    Study design and conductreasoning

    Nanoparticles were not given intravenously because the repeated injections would damage the tail veins, making it difficult to continue treatment through the entire period.

    “Of note, nanoparticles were not given intravenously because the repeated injections would damage the tail veins, making it difficult to continue treatment through the entire period.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    The experiments testing the effects of DNase I on invasion in vitro were repeated by three different co-authors.

    “The experiments testing the effects of DNase I on invasion in vitro were repeated by three different co-authors.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    The investigators who assessed, measured, or quantified the results of invasion through Matrigel, NET-like structures by CILI, or metastatic burden after treatment with DNase I-coated nanoparticles were blinded to the specific intervention.

    “The investigators who assessed, measured, or quantified the results of invasion through Matrigel, NET-like structures by CILI, or metastatic burden after treatment with DNase I-coated nanoparticles were blinded to the specific intervention.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    4T1 cells and C3(1)-Tag tumors express high levels of G-CSF.

    “Both 4T1 cells and C3(1)-Tag tumors express high levels of G-CSF (22, 35), and they are both models of triple-negative breast cancer.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [22] Park JH Rasch MG Qiu J Lund IK Egeblad M Presence of insulin-like growth factor binding proteins correlates with tumor-promoting effects of matrix metalloproteinase 9 in breast cancer Neoplasia 17 421 433 2015 26025665 PMC4468371 source

      Not checked in the free run.

    • cites [35] Kowanetz M Wu X Lee J Tan M Hagenbeek T Qu X Yu L Ross J Korsisaari N Cao T Bou-Reslan H Kallop D Weimer R Ludlam MJ Kaminker JS Modrusan Z van Bruggen N Peale FV Carano R Meng YG Ferrara N Granulocyte-colony stimulating factor promotes lung metastasis through mobilization of Ly6G+Ly6C+ granulocytes Proc Natl Acad Sci U S A 107 21248 21255 2010 21081700 PMC3003076 source

      Not checked in the free run.

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

    Study design and conductnone given

    Endpoints for animal experiments were selected before the conduction of the experiments as the time when the first animals in any experimental group had a weight loss of >10%.

    “Endpoints for animal experiments were selected before the conduction of the experiments as the time when the first animals in any experimental group had a weight loss of >10%.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    HMGB1 released during NET formation may drive proliferation and migration of cancer cells.

    “HMGB1 released during NET formation may drive proliferation and migration of cancer cells (12).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [12] Tohme S Yazdani HO Al-Khafaji AB Chidi AP Loughran P Mowen K Wang Y Simmons RL Huang H Tsung A Neutrophil Extracellular Traps Promote the Development and Progression of Liver Metastases after Surgical Stress Cancer Res 76 1367 1380 2016 26759232 PMC4794393 source

      Not checked in the free run.

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

    Interpretationreasoning

    Cancer cell expression of G-CSF could identify patients at risk of developing NET-promoted metastasis.

    “However, because we identified G-CSF as a critical factor in the induction of NETs by cancer cells, it is possible that cancer cell expression of G-CSF could identify patients at risk of developing NET-promoted metastasis.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: 4T1 cells and C3(1)-Tag tumors are both models of triple-negative breast cancer.

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

    Interpretationreasoning

    This concept for how cancer cells exploit host cells represents a therapeutic opportunity to prevent metastasis.

    “This concept for how cancer cells exploit host cells represents a therapeutic opportunity to prevent metastasis, which accounts for the great majority of cancer-associated deaths.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Metastasis accounts for the great majority of cancer-associated deaths.

  • Classified as probably not a substantial claim.

    Premisecitation

    Treatment with nucleases reduces metastasis.

    “Treatment with nucleases was reported to reduce metastasis decades ago, but with no insights as to why (36).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [36] Sugihara S Yamamoto T Tsuruta J Tanaka J Kambara T Hiraoka T Miyauchi Y Serine protease-induced enhancement of blood-borne metastasis of rat ascites tumour cells and its prevention with deoxyribonuclease Br J Cancer 62 607 613 1990 2121220 PMC1971483 source

      Not checked in the free run.

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

    Study design and conductnone given

    Animals were excluded from the study if the initial intravenous injection of the cancer cells missed the tail vein.

    “Animals were excluded from the study if the initial intravenous injection of the cancer cells missed the tail vein.”
    Quote in the paperNot a substantial claim

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

    Interpretationreasoning

    Mild neutropenia is associated with better survival because neutrophils play a functional role in metastasis.

    “However, our findings suggest an alternative hypothesis: that mild neutropenia is associated with better survival because neutrophils play a functional role in metastasis.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    NETs may stimulate the further invasion of the cells into the tissue and the expansion of the colonizing cells.

    “Instead, our data suggest that NETs may stimulate the further invasion of the cells into the tissue and the expansion of the colonizing cells.”
    Quote in the paperSubstantial claim

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

    Interpretationcitation

    Increased local vascular permeability would permit cancer cells to extravasate more easily.

    “which would permit cancer cells to extravasate more easily.”
    Quote in the paperNot a substantial claim

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

    Interpretationnone given

    The DNA mesh of NETs may promote invasion by acting to concentrate NET-associated proteases or protecting proteases from the endogenous inhibitors abundant in blood.

    “The DNA mesh of NETs may promote invasion by acting to concentrate NET-associated proteases or protecting proteases from the endogenous inhibitors abundant in blood.”
    Quote in the paperNot a substantial claim
40 of 40

  • The paper relies on this without support.

    The immunofluorescence staining used on human clinical samples detects NETs specifically.

    “To determine whether NETs could also be found in metastatic human breast cancer, we performed immunofluorescence staining on a small panel of clinical samples of primary tumor and matched metastatic lung lesions from breast cancer patients (Fig. 2A).”

  • The paper relies on this without support.

    The number of histologically detectable metastatic foci reflects expansion of disseminated cancer cells rather than initial seeding.

    “Unexpectedly, cancer cell-induced NETs did not promote extravasation but did influence the number of histologically detectable metastatic foci. This suggests that NETs mediate the expansion of disseminated cells.”

  • The paper relies on this without support.

    The mouse models used reflect human breast cancer metastasis.

    “To investigate whether neutrophils play a role in metastasis, we first compared neutrophil infiltration into tumors from orthotopically transplanted 4T1 and 4T07 murine breast cancer cells.”

  • The paper relies on this without support.

    DNase I-coated nanoparticles reduce metastasis specifically by digesting NETs.

    “Daily intraperitoneal injection of DNase I-coated nanoparticles significantly (p=0.002) reduced metastatic burden after intravenous injection of 4T1 cells (Figs. 6C, D).”

  • The paper relies on this without support.

    Cancer cells can acquire the ability to invade and migrate through many other means.

    “but cancer cells can acquire the ability to invade and migrate through many other means.”

  • The paper relies on this without support.

    4T1 cells and C3(1)-Tag tumors are both models of triple-negative breast cancer.

    “and they are both models of triple-negative breast cancer.”

  • The paper relies on this without support.

    Metastasis accounts for the great majority of cancer-associated deaths.

    “which accounts for the great majority of cancer-associated deaths.”
7 of 7

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Authors
Park, J., Wysocki, R. W., Amoozgar, Z., Maiorino, L., Fein, M. R., Jorns, J., Schott, A. F., Kinugasa-Katayama, Y., Lee, Y., Won, N. H., Nakasone, E. S., Hearn, S. A., Küttner, V., Qiu, J., Almeida, A. S., Perurena, N., Kessenbrock, K., Goldberg, M. S., & Egeblad, M.
Affiliations
  • Cold Spring Harbor Laboratory (US)
  • Dana-Farber Cancer Institute (US)
  • University of Michigan (US)
  • Cold Spring Harbor Laboratory, Cold Spring Harbor, NY 11724, USA.
  • Korea University Anam Hospital (KR)
  • St Giles Foundation (US)
  • University of California, Irvine (US)
  • Harvard University (US)
Corresponding author
  • Park, Juwon
  • Wysocki, Robert W.
  • Amoozgar, Zohreh
  • Maiorino, Laura
  • Perurena, Naiara
  • Egeblad, Mikala
ORCID
Venue
Science Translational Medicine
Publisher
American Association for the Advancement of Science (AAAS)
Published
2016-10-19
Volume
8
Issue
361
Pages
361ra138-361ra138
ISSN
  • 1946-6234 (ISSN-L)
  • 1946-6242
DOI
10.1126/scitranslmed.aag1711
PMC
PMC5550900
OpenAlex
W2534066964
Semantic Scholar
3ccd945ed1636147b459c8f2f8642742e2f3a425
Cited by
984
References
50
Citations by year
2026: 84 · 2025: 134 · 2024: 137 · 2023: 109 · 2022: 120 · 2021: 133 · 2020: 117 · 2019: 73 · 2018: 48 · 2017: 24 · 2016: 3 · 2015: 1
FWCI
29.92
Type
journal article
Language
en
Open access
Licence
www.sciencemag.org/about/science-licenses-journal-article-reuse (vor, 365 d)
Locations
  • Science Translational Medicine (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
  • Cold Spring Harbor Laboratory Institutional Repository (Cold Spring Harbor Laboratory) (submittedVersion)
  • eScholarship (California Digital Library) (submittedVersion)
Funders
  • CSHL Cancer Center Support (5P30CA045508)
  • U.S. Department of Defense (W81XWH-14-1-0078)
  • Long Island 2-Day Walk to Fight Breast Cancer
  • Joni Gladowsky Breast Cancer Foundation
  • NIH (5U01CA180944-02)
  • Hope Foundation
  • Cancer Research Institute
  • National Institute of General Medical Sciences Medical Scientist Training Program Training Award (T32-GM008444)
  • Aid for Cancer Research
  • Boehringer Ingelheim Fonds
  • Formación de Profesorado Universitario (AP2010-2197)
  • National Cancer Institute (K99 CA181490)
  • Deutsche Forschungsgemeinschaft (KU 3264/1-1)
Topics
  • Life Sciences › Immunology and Microbiology › Immunology › Neutrophil, Myeloperoxidase and Oxidative Mechanisms
  • Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
  • Health Sciences › Medicine › Oncology › Cancer Immunotherapy and Biomarkers
Keywords
neutrophil extracellular traps, breast cancer metastasis, tumor-host interaction, DNase I, lung metastasis, tumor invasion, tumor cell migration, triple-negative breast cancer, intravital imaging
MeSH terms
Animals, Cell Movement, Deoxyribonuclease I, Humans, Lung, Lung Neoplasms, Mice, Inbred BALB C, Neoplasm Metastasis, Neutrophils, Cell Line, Tumor, Mice, Nanoparticles, Triple Negative Breast Neoplasms, Extracellular Traps

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.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

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