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Cited by 203Extracellular vesicles in diseaseTelomeres, Telomerase, and SenescenceMesenchymal stem cell research

Small Extracellular Vesicles Have GST Activity and Ameliorate Senescence-Related Tissue Damage

Open access
Fafián-Labora, Juan Antonio, Rodríguez-Navarro, Jose Antonio, O’Loghlen, Ana·2020·Cell Metabolism
DOI 10.1016/j.cmet.2020.06.004PMC PMC7342013
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Abstract

Aging is a process of cellular and tissue dysfunction characterized by different hallmarks, including cellular senescence. However, there is proof that certain features of aging and senescence can be ameliorated. Here, we provide evidence that small extracellular vesicles (sEVs) isolated from primary fibroblasts of young human donors ameliorate certain biomarkers of senescence in cells derived from old and Hutchinson-Gilford progeria syndrome donors. Importantly, sEVs from young cells ameliorate senescence in a variety of tissues in old mice. Mechanistically, we identified sEVs to have intrinsic glutathione-S-transferase activity partially due to the high levels of expression of the glutathione-related protein (GSTM2). Transfection of recombinant GSTM2 into sEVs derived from old fibroblasts restores their antioxidant capacity. sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro. Altogether, our data provide an indication of the potential of sEVs as regenerative therapy in aging.

AI snapshot

Written from the paper’s full text.

TL;DR

Small extracellular vesicles from young human fibroblasts (sEV-Ys) ameliorate senescence biomarkers in old and Hutchinson-Gilford progeria syndrome cells and in old mice. sEV-Ys carry intrinsic glutathione-S-transferase activity, partly via GSTM2, increase reduced glutathione, and reduce oxidative stress and lipid peroxidation.

Key findings

  • sEV-Ys from young donor fibroblasts prevented proliferation arrest and reduced p16INK4A, p21CIP1, SA-β-Gal, and IL-8 in four old donor fibroblast lines (67–81 years) and four HGPS fibroblast lines (2–4 years); the soluble fraction had less effect and large EVs had no functionality.
  • Proteomic analysis identified 55 common proteins enriched in sEVs from control fibroblasts versus senescent sEVs, including glutathione conjugation pathway proteins; GSTM2 was highly enriched in young-cell sEVs and decreased in old/HGPS sEVs.
  • sEV-Ys had intrinsic GST activity, while soluble fractions did not; sEV-Y treatment increased GSH/GSSG in old cells, and BSO blunted the increase in proliferation and decrease in SA-β-Gal. Transfecting recombinant GSTM2 into old sEVs prevented SA-β-Gal activity.
  • In 22- to 25-month-old C57BL6 mice, intraperitoneal injection of 20 μg sEV-Ys twice a week for 3 weeks reduced SA-β-Gal in kidney, lung, and brown adipose tissue, decreased liver macrophage infiltration, lowered serum IL-6 and GM-CSF, reduced ROS in liver and serum, restored GST activity in liver and serum, and prevented MDA accumulation in liver and serum.

Methods

  • Primary human fibroblasts from young donors (GM05399, GM00969, GM05565, GM05758; 1–3 years), old donors (AG16086, AG06240, AG13152, AG13222; 67–81 years), and HGPS donors (AG11572, AG06917, AG07493, AG10677; 2–4 years) were treated with conditioned media fractions separated by serial ultracentrifugation into soluble fraction, large EVs, and small EVs.
  • In vitro analyses included BrdU incorporation, immunofluorescence for p16INK4A, p21CIP1, IL-8, 8-oxodG, p-γH2AX, SA-β-Gal, qPCR, immunoblotting, nanoparticle tracking analysis, GST activity assays, GSH/GSSG measurements, and MDA/TBARS assays.
  • In vivo, male C57BL/6 mice aged 22–25 months received 20 μg sEV-Ys or PBS intraperitoneally twice a week for 3 weeks; tissues and serum were analyzed for SA-β-Gal, senescence/SASP markers, ROS, GST activity, GSTM2, GSH, and lipid peroxidation. Statistics used Student’s t test unless otherwise specified, with ANOVA, Welch’s t test, or Mann-Whitney tests in some figures.

Limitations

  • The paper states that a caveat is the use of PBS as the control in old mice rather than sEV-Os, because sEV-Os might induce paracrine senescence in the few proliferating cells old mice have.
  • The paper states that it cannot exclude the possibility that additional components within sEV-Ys also contribute to preventing the senescent and aging phenotype, and that more experimental data and longer-term experiments would be needed before concluding sEV-Ys have rejuvenating potential.
  • Evident from the text: in vivo experiments used male mice only, with 3 young mice and 5 old mice per condition in several analyses; some tissue effects were trends or absent, such as no differences after subcutaneous sEV-Y injection except in serum, and little to no differences in lung for some SASP markers.

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

Integrity signals

Registry

Crossref · 10.1016/j.cmet.2020.06.004

Cites

Statistics

Nothing to recompute.

Claims the paper makes

0 of 39 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    sEVs noticeably prevented the cell-cycle arrest in old donor cells in comparison with the SF 11 days after treatment.

    “The sEVs noticeably prevented the cell-cycle arrest in old donor cells in comparison with the SF (Figure 1D) 11 days after treatment (day 8).”
    Quote in the paperSubstantial claim
  • Findingown data

    sEVs decrease lipid peroxidation both in vivo and in vitro.

    “sEVs increase the levels of reduced glutathione and decrease oxidative stress and lipid peroxidation both in vivo and in vitro.”
    Quote in the paperSubstantial claim
  • Findingown data

    iRAS cells treated with iCsEVs repress mRNA transcripts related to the SASP.

    “The heatmap in Figure 1H shows that iRAS cells treated with iCsEVs have the ability of repress mRNA transcripts related to the SASP and also cell-cycle inhibitors generally upregulated during senescence (Figure S1K).”
    Quote in the paperSubstantial claim
    • Rests on an assumption: sEVs from HFFF2 behave as sEV-Ys, and iCsEVs are equivalent to sEV-Ys while RASsEVs are equivalent to sEV-Os.
  • Findingown data

    The effect of SF was modest in comparison with sEVs.

    “Interestingly, although the soluble fraction (SF) also ameliorated the proliferation arrest characteristic of senescence, its effect was modest in comparison with sEVs while large EVs (lEVs) displayed no functionality.”
    Quote in the paperNot a substantial claim
  • Findingown data

    The lEV fraction had no effect on modifying senescence-related biomarkers in old donor cells.

    “We found the SF to have slightly less ability to ameliorate these senescence-related biomarkers in comparison with the sEV fraction, while the lEV fraction had no effect on modifying these biomarkers in old donor cells (Figures 1A–1C and S1C).”
    Quote in the paperSubstantial claim
  • May combine two claims. 28
  • Classified as probably not a substantial claim. 22
  • The quote only partly states this claim. 1

  • May combine two claims.

    Findingown data

    sEV-Ys prevent lipid peroxidation in vitro and in certain organs in mice of old age.

    “Furthermore, sEV-Ys have the ability to reverse the accumulation of reactive oxygen species (ROS) and prevent lipid peroxidation in vitro and in certain organs in mice of old age.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    sEVs and lEVs showed different EV distribution, mean particle size, and number by nanoparticle tracking analysis.

    “Next, we characterized the EV fraction and found that sEVs and lEVs showed a different EV distribution, mean particle size, and number by performing nanoparticle tracking analysis (NTA) (Figures S1D and S1E), while sEVs presented EV-related proteins such as ALIX and TSG101 (Figure S1F) and lacked endoplasmic reticulum contaminants such as calnexin (Figure S1G).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Treatment of four HGPS cells with pooled sEV-Ys prevented the proliferation arrest characteristic of these cells, with no effect observed from the SF at longer-term treatment.

    “Furthermore, treatment of four HGSP cells with pooled sEV-Ys prevented the proliferation arrest characteristic of these cells with no effect observed from the SF at longer-term treatment (Figure 2D).”
    Quote in the paperSubstantial claim

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

    Findingown data

    sEV particle size from HGPS cells is not changed compared to young donors.

    “As with the sEVs derived from old donors, we could also observe an increase in sEV number from HGPS cells compared to young donors by NTA with no change in particle size (Figures S2E and S2F).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    sEV-Y treatment reduced the mRNA expression of different SASP factors in the liver and kidney of old mice, with little to no differences in the lung.

    “we find a reduction in the mRNA expression of different SASP factors when old mice were treated with sEV-Ys in the liver and kidney with little to no differences in the lung (Figures 5E and S5B)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Only the sEV fraction had the potential to prevent ROS accumulation by quantifying 8-oxodG-positive cells, while the SF and lEV fractions had less effect.

    “Interestingly, we could confirm that only the sEV fraction had the potential to prevent ROS accumulation by quantifying 8-oxodG-positive cells, while the SF and lEV fractions had less effect (Figures 3G, 3H, and S3C).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    sEV-Y treatment reduced the levels of Cdkn1a mRNA in the liver and kidney, but not in the lung, of old mice.

    “we could only observe a reduction in the levels of Cdkn1a upon treatment in the liver and kidney, not in the lung (Figure S5A)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    MDA levels in the liver and serum increased in old mice compared to young mice, showing an increase in lipid peroxidation in old mice.

    “We could indeed observe significant changes in MDA levels in the liver and serum between young and old mice showing an increase in lipid peroxidation in old mice (Figures 7A and 7B)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Transfection of rGSTM2 into sEVs from old donors and treatment of old cells prevented SA-β-Gal activity.

    “when we transfected rGSTM2 into sEVs from old donors and treated old cells, we prevented SA-β-Gal activity (Figure 4I)”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Treatment of young cells with 4-HNE induces senescence, which is prevented by treatment with sEV-Ys.

    “treatment of young cells with 4-HNE induces senescence, which is prevented by treatment with sEV-Ys.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Increased transcriptional expression of several GSH synthesis enzymes upon sEV-Y treatment, in particular the GSH master regulator Gclc, boosts the intracellular GSH pool.

    “we observe increased transcriptional expression of several GSH synthesis enzymes upon sEV-Y treatment, in particular the GSH master regulator Gclc boosting the intracellular GSH pool.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    A decrease in the levels of MDA using these inhibitors and the implication of a DNA-damage response by analyzing the levels of p-γH2AX were further confirmed.

    “A decrease in the levels of MDA using these inhibitors and the implication of a DNA-damage response by analyzing the levels of p-γH2AX were further confirmed (Figures S7G and S7H)”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    GST needs the supply of reduced GSH, which we have shown to be extremely low in old cells and mice tissue.

    “GST needs the supply of reduced GSH, which we have shown to be extremely low in old cells and mice tissue.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    Cells undergoing senescence release a larger number of particles or sEVs as a characteristic of the SASP.

    “We and others have described that cells undergoing senescence release a larger number of particles or sEVs as a characteristic of the SASP (Borghesan et al., 2019, Takahashi et al., 2017).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Borghesan et al., 2019 source

      Not checked in the free run.

    • cites Takahashi et al., 2017 source

      Not checked in the free run.

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

    Study design and conductown data

    Longer-term experiments would be needed to confirm the sEV effect on cell-cycle arrest.

    “However, it cannot be excluded that longer-term experiments would be needed to confirm this.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    sEVs derived from young cells ameliorate different biomarkers of senescence, which can be further transmitted.

    “Altogether, our data show that sEVs derived from young cells ameliorate different biomarkers of senescence, which can be further transmitted.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: sEVs from HFFF2 behave as sEV-Ys, and iCsEVs are equivalent to sEV-Ys while RASsEVs are equivalent to sEV-Os.

  • May combine two claims.

    Interpretationown data

    sEV-Ys have intrinsic GST activity and can modulate GSH levels in recipient cells by regulating senescence in old cells.

    “Altogether, these data show that sEV-Ys have intrinsic GST activity and can modulate the GSH levels in recipient cells by regulating senescence in old cells.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationown data

    sEV-Ys can regulate ROS and GST activity with a concurrent increase in the antioxidant GSH and related mRNA transcripts in certain tissues in old mice.

    “Altogether, here we show that sEV-Ys can regulate ROS and GST activity with a concurrent increase in the antioxidant GSH and related mRNA transcripts in certain tissues in old mice.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Cell culture experiments used sEV-depleted FBS media to avoid FBS-derived sEV contamination.

    “All cells were washed twice with PBS prior to starting the experiments to remove excess FBS media and maintained in sEV-depleted FBS media for the duration of the experiment.”
    • Also stated in: § Method Details · plain
      “FBS was depleted of sEV by overnight (ON) ultracentrifugation at 100,000 g at 4C (Sorvall 100SE Ultracentrifuge).”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    Mice used in experiments were specific-pathogen free, with annual Full-FELASA health screening.

    “Mice were specific-pathogen free [Health screening (Full-FELASA profile) was performed annually] and maintained at the animal facility at the Hospital Ramón y Cajal of Madrid (Spain).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    When the majority of cells existing in the tissue are senescent cells, tissue homeostasis becomes compromised as there is transmission of paracrine senescence.

    “When the majority of cells existing in the tissue are senescent cells, the tissue homeostasis becomes compromised as there is transmission of paracrine senescence (Borghesan et al., 2019, Acosta et al., 2013)”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Borghesan et al., 2019 source

      Not checked in the free run.

    • cites Acosta et al., 2013 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    GSTM2 is contained within sEVs, conferring upon them GST antioxidant activity, which ameliorates senescence, in contrast to the SF fraction.

    “we show that GSTM2 is contained within sEVs, conferring upon them GST antioxidant activity, which ameliorates senescence, in contrast to the SF fraction.”
    Quote in the paperSubstantial claim

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

    Study design and conductnone given

    For high content analysis, multiple fields per well were acquired to include a minimum of 100 cells per sample well.

    “Multiple fields per well were acquired to include a minimum of 100 cells per sample well.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    qPCR data were normalized to a housekeeping gene, RPS14, except for Taqman probes.

    “The data were normalized to a housekeeping gene, RPS14, except for the Taqman probes.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Both SF and sEVs can mediate paracrine senescence, yet lEVs had no functionality.

    “We recently showed that both SF and sEVs can mediate paracrine senescence, yet lEVs had no functionality (Borghesan et al., 2019)”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Borghesan et al., 2019 source

      Not checked in the free run.

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

    Study design and conductnone given

    A threshold for positive cells in immunofluorescence was assigned above the average intensity of unstained or negative control samples.

    “In samples of cultured cells, a threshold for positive cells was assigned above the average intensity of unstained or negative control samples.”
    Quote in the paperNot a substantial claim

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

    Study design and conductown data

    All relevant data of the experiments were submitted to the EV-TRACK knowledgebase (EV-TRACK ID: EV200036).

    “We have submitted all relevant data of our experiments to the EV-TRACK knowledgebase (EV-TRACK ID: EV200036) (Van Deun et al., 2017).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Van Deun et al., 2017 source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    One of the main substrates of GSTM2 are lipids.

    “One of the main substrates of GSTM2 are lipids.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Townsend and Tew, 2003

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    GSTM2-contained sEVs are a limiting factor and are the initial trigger of the antioxidant pathway, which needs to be maintained transcriptionally to see the antioxidant long-term response observed in vivo.

    “As GSTM2-contained sEVs are a limiting factor, we observe they are the initial trigger of the antioxidant pathway, which needs to be maintained transcriptionally to see the antioxidant long-term response we observe in vivo.”
    Quote in the paperSubstantial claim

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

    Study design and conductreasoning

    Use of PBS as a control in old mice in comparison with sEV-Ys injected into old mice is a caveat of the current study.

    “One caveat of the current study is the use of PBS as a control in old mice in comparison with sEV-Ys injected into old mice.”
    Quote in the paperNot a substantial claim
    • The authors state this as an assumption: sEV-Os might induce paracrine senescence in the few proliferating cells that old mice have.

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

    Study design and conductreasoning

    Conditioned media was only used once per individual experiment.

    “All cells were plated for each individual experiment and thus the conditioned media was only used once.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    eNAMPT is present within EVs to ameliorate aging.

    “nicotinamide phosphoribosyltransferase (eNAMPT) was shown to be present within EVs to ameliorate aging.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Yoshida et al., 2019 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    sEV-Ys ameliorate senescence in old recipient cells and old mice.

    “In this study, we are describing that sEV-Ys ameliorate senescence in old recipient cells and old mice.”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The oxidation of lipids induces degradation products through lipid peroxidation, generating 4-HNE and MDA, which can modify protein function.

    “The oxidation of lipids induces degradation products through a process known as lipid peroxidation. This results in the generation of highly reactive products such as reactive aldehydes 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA), which can modify protein function (Binder et al., 2016).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Binder et al., 2016 source

      Not checked in the free run.

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

    Study design and conductnone given

    Flow cytometry gates were set using NovoExpress Software to analyze single-cell fluorescence.

    “Gates were set using the NovoExpress Software to analyze single cells fluorescence.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    GSTM2 is a cytosolic enzyme that catalyzes the conjugation of GSH aimed at the elimination of ROS-generated toxic byproducts.

    “GSTM2 is a cytosolic enzyme that catalyzes the conjugation of GSH aimed at the elimination of ROS-generated toxic byproducts (Townsend and Tew, 2003)”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Townsend and Tew, 2003

      Not checked in the free run.

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

    Premisecitation

    eNAMPT-EVs can induce NAD+ in recipient cells and counteract aging.

    “eNAMPT-EVs can induce NAD+ in recipient cells and counteract aging (Yoshida et al., 2019)”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Yoshida et al., 2019 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    There is a crosstalk between both cell types via EVs; EVs inducing senescence in young cells and EVs preventing senescence in old cells.

    “Thus, there seems to be a crosstalk between both cells types via EVs; EVs inducing senescence in young cells and EVs preventing senescence in old cells.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: The tissue holds a mixture of senescent and proliferating cells.
    • Rests on an assumption: The temporary presence of senescent cells is generally beneficial for an individual while the accumulation is detrimental.

  • May combine two claims.

    Interpretationcitation

    During earlier stages of aging or tissue damage, when there are still plenty of proliferating cells, these can 'repair' tissue dysfunction by ameliorating the senescent phenotype of damaged cells through soluble factors and via sEVs.

    “during the earlier stages of aging or during tissue damage, when there are still plenty of proliferating cells, these can “repair” tissue dysfunction by ameliorating the senescent phenotype of damaged cells through soluble factors (Mosteiro et al., 2016, Ocampo et al., 2016) and via sEVs as shown in this study.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites Mosteiro et al., 2016 source

      Not checked in the free run.

    • cites Ocampo et al., 2016 source

      Not checked in the free run.

    • Rests on an assumption: The tissue holds a mixture of senescent and proliferating cells.

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

    Premisecitation

    EVs play a key role in maintaining tissue homeostasis.

    “EVs also play a key role in maintaining tissue homeostasis (Kalluri and LeBleu, 2020, Lee and Schmitt, 2019, van Niel et al., 2018)”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites Kalluri and LeBleu, 2020 source

      Not checked in the free run.

    • cites Lee and Schmitt, 2019 source

      Not checked in the free run.

    • cites van Niel et al., 2018 source

      Not checked in the free run.

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

    Premisecitation

    Accumulation of 4-HNE has been previously reported in senescence, aging, and age-related diseases.

    “accumulation of 4-HNE has been previously reported in senescence, aging, and age-related diseases (Ademowo et al., 2017, Jurk et al., 2014)”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Ademowo et al., 2017 source

      Not checked in the free run.

    • cites Jurk et al., 2014 source

      Not checked in the free run.

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

    Study design and conductnone given

    The NanoSight LM10 instrument was calibrated using silica microsphere beads.

    “The NanoSight LM10 (Malvern Instruments) was calibrated using Silica Microspheres beads.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    EVs have asparaginase and succinase activity to suppress neuroinflammation.

    “EVs have been shown to have asparaginase and succinase activity to suppress neuroinflammation (Iraci et al., 2017, Peruzzotti-Jametti et al., 2018)”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Iraci et al., 2017 source

      Not checked in the free run.

    • cites Peruzzotti-Jametti et al., 2018 source

      Not checked in the free run.

43 of 43

  • The paper relies on this without support.

    EVs do not induce toxicity or immunogenicity.

    “Instead, EVs are a natural source and do not induce toxicity or immunogenicity; thus, this study opens new avenues for strategies to improve lifelong health and wellbeing.”

  • The paper relies on this without support.

    sEVs from HFFF2 behave as sEV-Ys, and iCsEVs are equivalent to sEV-Ys while RASsEVs are equivalent to sEV-Os.

    “HFFF2 are derived from 14- to 18-week-old human fetus and their sEVs behave as sEV-Ys.”
    “iRAS cells treated with sEVs from either iC (iCsEVs, equivalent to sEV-Ys) or iRAS (RASsEVs, equivalent to sEV-Os)”

  • The paper relies on this without support.

    The tissue holds a mixture of senescent and proliferating cells.

    “It is known that the tissue holds a mixture of senescent and proliferating cells.”

  • The paper relies on this without support.

    The temporary presence of senescent cells is generally beneficial for an individual while the accumulation is detrimental.

    “In fact, the temporary presence of senescent cells is generally beneficial for an individual while the accumulation is detrimental.”

  • The authors state this as an assumption.

    The predominance of functionality between sEV-Ys and sEV-Os depends on the proportion of each cell present in the tissue.

    “We believe that the predominance of functionality between sEV-Ys and sEV-Os will depend on the proportion of each cell present in the tissue.”

  • The authors state this as an assumption.

    sEV-Os might induce paracrine senescence in the few proliferating cells that old mice have.

    “We hypothesized, based on our previous findings (Borghesan et al., 2019), that sEV-Os might induce paracrine senescence in the few proliferating cells that old mice have.”
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?

  • Small extracellular vesicles enriched in GSTM2 can relieve senescence in various tissues when injected intraperitoneally in old mice.

    “Therapeutically, small extracellular vesicles enriched in the glutathione S-transferase Mu 2 (GSTM2) enzyme, which works in conjunction with GSH to reduce OS and detoxify the cell from harmful compounds, can relieve senescence in various tissues when injected intraperitoneally in old mice [190].”

    From The Role of Antioxidants in the Interplay between Oxidative Stress and Senescence (2022) · cites this paper as [190]

Self-check

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

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

  • GM05565 is labeled both old and young

    Minor

    The Figure 3 legend calls GM05565 an old cell line, but the cell-culture methods list it among the young-derived fibroblasts.

1 of 1

The citation lists haven’t been fetched yet

References 44 · Cited by 203

Research works printed but not linked: 1

  1. Kuilman, 2010, The essence of senescence, Genes Dev., 24, 2463, 10.1101/gad.1971610 · DOI · Paper page

Authors
Fafián-Labora, J. A., Rodríguez-Navarro, J. A., & O’Loghlen, A.
Affiliations
Queen Mary University of London (GB)
Corresponding author
O’Loghlen, Ana
ORCID
O’Loghlen, Ana
Venue
Cell Metabolism
Publisher
Elsevier BV
Published
2020-07
Volume
32
Issue
1
Pages
71-86.e5
ISSN
1550-4131 (ISSN-L)
DOI
10.1016/j.cmet.2020.06.004
PMC
PMC7342013
OpenAlex
W3036058245
Cited by
203
References
44
Citations by year
2026: 26 · 2025: 39 · 2024: 40 · 2023: 33 · 2022: 31 · 2021: 28 · 2020: 6
FWCI
8.02
Type
journal article
Language
en
Open access
Licence
Locations
  • Cell Metabolism (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • LA Referencia (Red Federada de Repositorios Institucionales de Publicaciones Científicas) (submittedVersion, cc-by)
  • RUC (Universidade Da Coruña) (submittedVersion, cc-by)
Funders
  • Ministerio de Economía y Competitividad (SAF2016-78666R)
  • Xunta de Galicia (ED481B 2017/117)
  • Biotechnology and Biological Sciences Research Council (BB/P000223/1, MGU0497)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Extracellular vesicles in disease
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Health Sciences › Medicine › Genetics › Mesenchymal stem cell research
Keywords
small extracellular vesicles, cellular senescence, aging, glutathione S-transferase, GSTM2, Hutchinson-Gilford progeria syndrome, oxidative stress, lipid peroxidation, reduced glutathione, antioxidant capacity, regenerative therapy, fibroblasts, tissue damage
MeSH terms
Extracellular Vesicles, Aging, Animals, Glutathione Transferase, Humans, Male, Mice, Inbred C57BL, Cellular Senescence, Mice

About these checks

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

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-06.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-06.

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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