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Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps
Open accessAbstract
Treatment with DNase I–coated nanoparticles prevents metastasis by targeting neutrophil extracellular traps induced by cancer cells in a mouse model.
Overview
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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.
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Claims the paper makes
67 Own result · 27 Own interpretation · 22 Not checked yet · 7 Own method · 1 No basis given
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Abstract
plaindirectn = 3- Asserted: passes — The authors put it forward as true.
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- Rests on an assumption: DNase I-coated nanoparticles reduce metastasis specifically by digesting NETs.
- Also stated in: § DNase I treatment prevents lung metastasis in mice · plain
The paper states this without a source or data behind it here, so there is nothing to check it against. The quote shows it is in the paper.
In the paper
“Consistent with this idea, CXCL1/2 knockdown did not increase primary tumor growth in T cell-deficient, nude mice (Fig. 1E).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Abstract
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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- Also stated in: § NETs promote cancer cell migration and invasion · plain
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“However, CXCL1/2 knockdown did significantly (p=0.03) reduce spontaneous metastasis in nude mice (Fig. 1F).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaindirect- Asserted: passes — The authors put it forward as true.
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In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaincorrelational- Asserted: passes — The authors put it forward as true.
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In the paper
“Tumors with reduced expression of CXCL1/2 had an approximately doubled tumor burden, but, despite this, metastatic burden was not increased (fig. S1).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaincorrelational- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“This increased migration was blocked by DNase I treatment (Fig. 3H).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > NETs promote cancer cell migration and invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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In the paper
“Instead, metastatic burden from CXCL1/2 knockdown cells was significantly (p=0.0008) decreased when equal numbers of cancer cells were injected intravenously into mice (Fig. 1D).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“C3(1)-Tag cells have a limited ability to invade through Matrigel, but neutrophils significantly (p=0.047) increased the migration of C3(1)-Tag cells across uncoated membranes.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > NETs promote cancer cell migration and invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Tumors with reduced expression of CXCL1/2 had an approximately doubled tumor burden, but, despite this, metastatic burden was not increased (fig. S1).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
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- It may state two separate claims as one. 25
- Probably not a substantial claim. 19
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Abstract
plaindirect- Asserted: passes — The authors put it forward as true.
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- Also stated in: § NETs promote cancer cell migration and invasion · plain
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plain- Asserted: passes — The authors put it forward as true.
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In the paper
“We detected NETs in 16 out of 20 primary tumors, and in 13 out of 19 metastatic lung lesions (Fig. 2B).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plain- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
- Rests on an assumption: The immunofluorescence staining used on human clinical samples detects NETs specifically.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
boosteddirect- Asserted: passes — The authors put it forward as true.
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In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaincorrelational- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Co-culturing with neutrophils increased the invasion of 4T1 cells, but had little effect on the invasion of 4T07 cells (Fig. 3C).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > NETs promote cancer cell migration and invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: passes — The paper’s conclusions depend on it.
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It may state two separate claims as one.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > NETs promote cancer cell migration and invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Metastatic cancer cells induce formation of NETs
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Rests on an assumption: The immunofluorescence staining used on human clinical samples detects NETs specifically.
A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > DNase I treatment prevents lung metastasis in mice
plainn = 3- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In addition, the CM lost its ability to stimulate invasion if DNase I was added after collection or upon heat denaturing (fig. S6).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > DNase I treatment prevents lung metastasis in mice
plain- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
plaindirect- Asserted: passes — The authors put it forward as true.
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A result the paper reports from its own data — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > DNase I treatment prevents lung metastasis in mice
plaindirectn = 6- Asserted: passes — The authors put it forward as true.
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In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > DNase I treatment prevents lung metastasis in mice
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
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What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Unfortunately, the p47phox-/- strain is on the C57BL/6 background, precluding in vivo experiments with the BALB/c-derived 4T1 cells.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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- Load-bearing: fails — The paper’s conclusions may not depend on it.
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Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. It may state two separate claims as one.
In the paper
“NETs that form intravascularly in response to infection can promote extravasation of disseminated cancer cells (5).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > DNase I treatment prevents lung metastasis in mice
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
What the sources say
- Not checked yetcites [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
plain- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
In the paper
“G-CSF can prime neutrophils for NET formation and is secreted by 4T1 cells (23).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > NETs promote cancer cell migration and invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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It may state two separate claims as one.
In the paper
“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).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
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Probably not a substantial claim.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Thus, metastatic cancer cells activate signaling pathways that include NADPH oxidase and PAD4 in neutrophils, causing the formation of invasion-promoting NETs.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
plaindirect- Asserted: passes — The authors put it forward as true.
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None of the sources cited for this claim have been checked yet.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Results > Cancer cells induce lytic NETs, and blocking NET formation reduces invasion
What the sources say
- Not checked yetcites [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 yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
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Probably not a substantial claim.
In the paper
“In addition, intravascular NETs can increase local vascular permeability (15), which would permit cancer cells to extravasate more easily.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Materials and Methods > In vivo treatment with DNase I-coated nanoparticles
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“The experiments testing the effects of DNase I on invasion in vitro were repeated by three different co-authors.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Materials and Methods > Study design
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Materials and Methods > Study design
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
None of the sources cited for this claim have been checked yet.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
What the sources say
- Not checked yetcites [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 yetcites [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
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“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%.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Materials and Methods > Study design
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
In the paper
“HMGB1 released during NET formation may drive proliferation and migration of cancer cells (12).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
What the sources say
- Not checked yetcites [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
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Rests on an assumption: 4T1 cells and C3(1)-Tag tumors are both models of triple-negative breast cancer.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
plain- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may be too vague to check against evidence.
- Rests on an assumption: Metastasis accounts for the great majority of cancer-associated deaths.
In the paper
“Treatment with nucleases was reported to reduce metastasis decades ago, but with no insights as to why (36).”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
What the sources say
- Not checked yetcites [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
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
What the paper did — it cites no source for this, so there is none to check. The quote shows it is in the paper.
In the paper
“Animals were excluded from the study if the initial intravenous injection of the cancer cells missed the tail vein.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Materials and Methods > Study design
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“However, our findings suggest an alternative hypothesis: that mild neutropenia is associated with better survival because neutrophils play a functional role in metastasis.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Instead, our data suggest that NETs may stimulate the further invasion of the cells into the tissue and the expansion of the colonizing cells.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“which would permit cancer cells to extravasate more easily.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“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.”
§ Cancer cells induce metastasis-supporting neutrophil extracellular DNA traps > Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
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.”
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?
In the citing paper
“recently, through increasing evidence, NETs have been directly associated with the initiation and induction of tumor invasion, aiding metastasis [20, 33–36].”
Citations
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The Temporal Landmarks of Cancer
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IRCI-2025 bridges global expertise to accelerate progress in cancer and infection immunology
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Metadata
- 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
- Published
- 2016-10-19
- Volume
- 8
- Issue
- 361
- Pages
- 361ra138-361ra138
- ISSN
- 1946-6234 (ISSN-L)
- 1946-6242
- 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
- FWCI
- 29.92
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- 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.
A language model drew the claims from the paper on 2026-10-10, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-10 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
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.
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