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Cited by 206Telomeres, Telomerase, and SenescenceProtein Degradation and InhibitorsGenetics, Aging, and Longevity in Model Organisms

A BET family protein degrader provokes senolysis by targeting NHEJ and autophagy in senescent cells

Open access
Wakita, Masahiro, Takahashi, Akiko, Sano, Osamu, Loo, Tze Mun, Imai, Yoshinori, Narukawa, Megumi, Iwata, Hidehisa, Matsudaira, Tatsuyuki +4 more·2020·Nature Communications
DOI 10.1038/s41467-020-15719-6PMC PMC7176673
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Abstract

Although cellular senescence acts primarily as a tumour suppression mechanism, the accumulation of senescent cells in vivo eventually exerts deleterious side effects through inflammatory/tumour-promoting factor secretion. Thus, the development of new drugs that cause the specific elimination of senescent cells, termed senolysis, is anticipated. Here, by an unbiased high-throughput screening of chemical compounds and a bio-functional analysis, we identify BET family protein degrader (BETd) as a promising senolytic drug. BETd provokes senolysis through two independent but integrated pathways; the attenuation of non-homologous end joining (NHEJ), and the up-regulation of autophagic gene expression. BETd treatment eliminates senescent hepatic stellate cells in obese mouse livers, accompanied by the reduction of liver cancer development. Furthermore, the elimination of chemotherapy-induced senescent cells by BETd increases the efficacy of chemotherapy against xenograft tumours in immunocompromised mice. These results reveal the vulnerability of senescent cells and open up possibilities for its control.

AI snapshot

Written from the paper’s full text.

TL;DR

An unbiased high-throughput screen of ~47,000 compounds identified BET family protein degraders/inhibitors as senolytic agents, with ARV825 showing strong senolysis at 5–10 nM. ARV825 kills senescent cells by inhibiting NHEJ and activating autophagy, reducing obesity-associated HCC and improving chemotherapy efficacy in mouse models.

Key findings

  • Screening ~47,000 small molecules found 15 compounds that preferentially killed Ras-induced senescent cells; four top hits were BET inhibitors (JQ1, I-BET151, I-BET762, PFI-1). ARV825, a BET degrader, had the strongest senolytic activity, killing senescent HDFs at 5–10 nM regardless of senescence induction method, and reduced BRD3/BRD4 levels at 10 nM.
  • In an obesity-induced HCC mouse model, ARV825 (5 mg/kg i.p.) substantially reduced HCC development and reduced senescent hepatic stellate cells, judged by αSMA with p21, IL-6, or Groα staining (HFD+vehicle n=20; HFD+ARV825 n=16).
  • In HCT116 xenografts, DXR plus ARV825 significantly reduced tumour size versus DXR alone, with reduced p21 and 53BP1 and increased γH2AX, LC3B, and cleaved caspase-3 in tumour sections.
  • Mechanistically, ARV825 downregulated xrcc4, blocked 53BP1 recruitment to DSB sites, exacerbated DSBs, and upregulated autophagic genes; autophagy inhibitors bafilomycin A1 or chloroquine reduced ARV825-induced senolysis, and autophagy preceded caspase-dependent apoptosis.

Methods

  • HTS: IMR90-ER:HRAS cells were treated with 500 nM 4-hydroxytamoxifen for 7 days, plated at 500 cells/well in 384-well plates, incubated with ~47,000 Takeda compounds at 3 or 10 μM for 72 h, then assayed with CellTiter-Glo and Caspase-Glo3/7.
  • Cell and in vivo models: TIG-3 HDFs were used as early-passage controls (<46 population doublings) or replicative senescent cells (>70 population doublings); senescence was also induced with doxorubicin (250 ng/ml for TIG-3, 200 ng/ml for HCT116) or RasV12. Mouse models included HFD-fed obesity-associated HCC mice treated with ARV825 (5 mg/kg i.p.) and HCT116 xenografts treated with DXR (1 mg/kg daily for 7 days) followed by ARV825 (5 mg/kg, 10 times over 2 weeks).
  • Main analyses: RNA-seq, immunoblotting, immunoprecipitation, immunofluorescence/immunohistochemistry, RT-qPCR, neutral comet assay, cell proliferation/apoptosis assays, and standard statistical tests (t-test, ANOVA with Holm-Sidak or Tukey, Kruskal-Wallis with Dunn’s).

Limitations

  • The paper states that not all aspects of BRD4 inhibition-induced senolysis may be explained by the described factors; it remains unclear whether BETd senolytic effects are common to all senescent cell types in vivo, partly due to lack of absolute senescence markers. It also cautions that healthy non-dividing cells rely on NHEJ and that high ARV825 concentrations (>50 nM) reduced control cell proliferation, so optimal in vivo dosing is crucial.
  • Evident from the text: XRCC4 overexpression failed to rescue ARV825-induced DSBs and senolysis, and NU7026 alone increased DSBs without inducing senolysis, indicating additional mechanisms beyond NHEJ inhibition. The study relies on cultured cells and mouse models, with no human clinical data.

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

Integrity signals

Registry

Crossref · 10.1038/s41467-020-15719-6

Cites

53 of 53 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 38 cited sources checked · 0 supported · 0 flagged

  • Findingown data

    A high-throughput screen of about 47,000 small molecules identified 15 small molecules that preferentially induce cell death in Ras-induced senescent cells compared with control non-senescent cells.

    “A cell viability analysis based on measurements of the ATP levels and Caspase3/7 activity revealed that 15 small molecules preferentially induce cell death in Ras-induced senescent cells, as compared to the control non-senescent cells.”
    Quote in the paperSubstantial claim
  • Findingcitation

    Four top-ranking hit molecules from the screen, JQ1, I-BET151, I-BET762, and PFI-1, are inhibitors of BET family proteins.

    “four top ranking hit molecules (JQ1, I-BET151, I-BET762, and PFI-1) turned out to be inhibitors of bromodomain and extra-terminal (BET) family proteins21,23”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [21] Shu, S. & Polyak, K. BET bromodomain proteins as cancer therapeutic targets. Cold Spring Harb. Sympo. Quant. Biol. 81, 123–129 (2016). source

      Not checked in the free run.

    • cites [23] Abedin, S. M., Boddy, C. S. & Munshi, H. G. BET inhibitors in the treatment of hematologic malignancies: current insights and future prospects. OncoTargets Ther. 9, 5943–5953 (2016). source

      Not checked in the free run.

  • Findingown data

    Cultured senescent HCT116 cells provoked by doxorubicin were preferentially killed by ARV825 compared with control HCT116 cells.

    “cultured senescent HCT116 cells (a human colon cancer cell line) provoked by doxorubicin (DXR) treatment were preferentially killed by the ARV825 treatment, as compared to control HCT116 cells”
    Quote in the paperSubstantial claim
  • Findingown data

    Expression of a series of autophagic genes was upregulated by ARV825 treatment in senescent HDFs.

    “the expression of a series of autophagic genes was upregulated by the ARV825 treatment in senescent HDFs”
    Quote in the paperSubstantial claim
  • Findingown data

    Treatment of senescent HDFs with autophagy inhibitors bafilomycin A1 or chloroquine substantially reduced ARV825-induced senolysis.

    “the treatment of senescent HDFs with autophagy inhibitors, such as bafilomycin A1 or chloroquine, substantially reduced the ARV825-induced senolysis”
    Quote in the paperSubstantial claim
  • May combine two claims. 22
  • Classified as probably not a substantial claim. 12

  • May combine two claims.

    Findingown data

    Co-treatment with NU7026 and cationic lipids provoked increases of LC3 puncta and senolysis in senescent HDFs.

    “the co-treatment with NU7026 and cationic lipids provoked increases of LC3 puncta and senolysis in senescent HDFs”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    The interaction between BRD4 and 53BP1 was observed in senescent cells and was reduced by ARV825 treatment.

    “the interaction between BRD4 and 53BP1 was observed in senescent cells, but was reduced when senescent HDFs were treated with ARV825”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Long-term ABT263 treatment caused apoptotic cell death in confluence-induced quiescent cells even at 500 nM, whereas ARV825 treatment did not.

    “although a long term treatment with ABT263 caused apoptotic cell death in confluence (contact-inhibition)-induced reversible cell-cycle arrest (quiescence), even at an optimal concentration for senolysis (500 nM), this was not the case with the ARV825 treatment”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    siRNA-based depletion of XRCC4 exacerbated DNA double-strand breaks without blocking 53BP1 localisation to DSB sites in senescent cells.

    “the siRNA-based depletion of XRCC4 exacerbated the formation of DSBs without blocking the 53BP1 localisation to DSB sites in senescent cells”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    53BP1 foci formation was abolished upon ARV825 treatment or BRD4 depletion, accompanied by exacerbation of DNA double-strand breaks in senescent HDFs.

    “53BP1 foci formation was abolished upon ARV825 treatment or BRD4 depletion, accompanied by the exacerbation of DSBs in senescent HDFs”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    ARV825 had the strongest senolytic activity among tested drugs and killed senescent HDFs at 5 to 10 nM regardless of the manner of senescence induction.

    “ARV825 had the strongest senolytic activity and killed senescent HDFs at a five to ten nanomolar concentration, regardless of the manner of cellular senescence induction”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    NU7026 treatment failed to induce senolysis in senescent HDFs although DNA double-strand breaks increased to the same levels as with ARV825 treatment.

    “the NU7026 treatment failed to induce senolysis in senescent HDFs, although the DSBs were increased to the same levels as those seen with the ARV825 treatment”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Separate treatment with either cationic lipids or the DNA-PK inhibitor did not provoke increases of LC3 puncta and senolysis in senescent HDFs.

    “this was not the case when senescent HDFs were separately treated with either the cationic lipids or the DNA-PK inhibitor”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Bafilomycin A1 treatment attenuated ARV825-induced senolysis, accompanied by a substantial reduction of activated caspase 3 levels.

    “On the other hand, although the bafilomycin A1 treatment attenuated the ARV825-induced senolysis, this was accompanied by a substantial reduction of the activated caspase 3 levels (Fig. 7a and c).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Treatment with the caspase inhibitor Q-VD-OHT attenuated the senolysis provoked by ARV825, without reducing the number of LC3 puncta in senescent HDFs.

    “and a treatment with a caspase inhibitor (Q-VD-OHT) attenuated the senolysis provoked by ARV825, without reducing the number of LC3 puncta in senescent HDFs (Fig. 7a and b).”
    Quote in the paperSubstantial claim

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

    Premisecitation

    The combination of dasatinib and quercetin exhibited remarkable cell-type specificity.

    “the combination of dasatinib and quercetin (D + Q), another previously reported senolytic drug20, apparently exhibited remarkable cell-type specificity15”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [15] Childs, B. G. et al. Senescent cells: an emerging target for diseases of ageing. Nat. Rev. Drug Discov. 16, 718–735 (2017). source

      Not checked in the free run.

    • cites [20] Zhu, Y. et al. The Achilles’ heel of senescent cells: from transcriptome to senolytic drugs. Aging Cell 14, 644–658 (2015). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    ARV825 has the potential to eliminate senescent hepatic stellate cells in vivo and thereby reduce HCC development, at least in the obesity mouse model.

    “strongly suggest that ARV825 has the potential to eliminate senescent HSCs in vivo, thereby reducing HCC development, at least in our obesity mouse model.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    The senescence-associated secretory phenotype exhibits deleterious side effects such as chronic inflammation and/or tumourigenesis depending on biological context.

    “it also exhibits deleterious side effects such as chronic-inflammation and/or tumourigenesis, depending on the biological context2,3,11–13.”
    Quote in the paperSubstantial claim0 of 5 sources checked
    • cites [2] Chan, A. S. L. & Narita, M. Short-term gain, long-term pain: the senescence life cycle and cancer. Genes Dev. 33, 127–143 (2019). source

      Not checked in the free run.

    • cites [3] Lee, S. & Schmitt, C. A. The dynamic nature of senescence in cancer. Nat. Cell Biol. 21, 94–101 (2019). source

      Not checked in the free run.

    • cites [11] Rodier, F. & Campisi, J. Four faces of cellular senescence. J. Cell Biol. 192, 547–556 (2011). source

      Not checked in the free run.

    • cites [12] Baker, D. J. et al. Naturally occurring p16Ink4a-positive cells shorten healthy lifespan. Nature 530, 184–189 (2016). source

      Not checked in the free run.

    • cites [13] Yoshimoto, S. et al. Obesity-induced gut microbial metabolite promotes liver cancer through senescence secretome. Nature 499, 97–101 (2013). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    ARV825 has senolytic activity in vivo and could have a beneficial impact on deleterious side effects of senescent cells depending on biological context.

    “Collectively, these results strongly suggest that ARV825 also has senolytic activity in vivo and could have some beneficial impact on the deleterious side effects of senescent cells, depending on the biological context.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    In a phase II study of ABT263 for advanced and recurrent small-cell lung carcinoma, transient thrombocytopenia and neutropenia were reported as side-effects.

    “in a phase II study of ABT263 (a specific inhibitor of the anti-apoptotic proteins BCL2 and BCL-xL)19 for the treatment of advanced and recurrent small-cell lung carcinoma patients, transient thrombocytopenia and neutropenia were reported as side-effects15.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [15] Childs, B. G. et al. Senescent cells: an emerging target for diseases of ageing. Nat. Rev. Drug Discov. 16, 718–735 (2017). source

      Not checked in the free run.

    • cites [19] Chang, J. et al. Clearance of senescent cells by ABT263 rejuvenates aged hematopoietic stem cells in mice. Nat. Med 22, 78–83 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    BETd provokes senolysis through two independent but integrated pathways: attenuation of non-homologous end joining and up-regulation of autophagic gene expression.

    “BETd provokes senolysis through two independent but integrated pathways; the attenuation of non-homologous end joining (NHEJ), and the up-regulation of autophagic gene expression.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Elimination of therapy-induced senescent cells reduced several side-effects of chemotherapy and cancer recurrence in mice.

    “the elimination of therapy-induced senescent cells reduced several side-effects of chemotherapy and even cancer recurrence in mice14.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [14] Demaria, M. et al. Cellular senescence promotes adverse effects of chemotherapy and cancer relapse. Cancer Disco. 7, 165–176 (2017). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Senolytic drugs identified to date were not discovered by a truly unbiased high-throughput screening method and appear to have limitations in clinical applications.

    “However, the senolytic drugs identified to date were not discovered by a truly unbiased high-throughput screening (HTS) method, and thus appear to have limitations in clinical applications15,17–19.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [15] Childs, B. G. et al. Senescent cells: an emerging target for diseases of ageing. Nat. Rev. Drug Discov. 16, 718–735 (2017). source

      Not checked in the free run.

    • cites [17] Yosef, R. et al. Directed elimination of senescent cells by inhibition of BCL-W and BCL-XL. Nat. Commun. 7, 11190 (2016). source

      Not checked in the free run.

    • cites [18] Baar, M. P. et al. Targeted apoptosis of senescent cells restores tissue homeostasis in response to chemotoxicity and aging. Cell 169, 132–147.e116 (2017). source

      Not checked in the free run.

    • cites [19] Chang, J. et al. Clearance of senescent cells by ABT263 rejuvenates aged hematopoietic stem cells in mice. Nat. Med 22, 78–83 (2016). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    Treatment with cationic lipids and exacerbation of DNA double-strand breaks are both required for full activation of the autophagic machinery in senescent cells.

    “indicating that the treatment with cationic lipids and the exacerbation of DSBs are both required for the full activation of the autophagic machinery in senescent cells.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductreasoning

    A combination of other senescence markers, including p21 induction, Lamin B1 reduction, and 53BP1 foci formation, was used throughout the study.

    “we decided to use a combination of other senescence markers, such as the induction of p21Waf1/Cip1/Sdi1 expression28, the reduction of Lamin B1 expression29,30, and the increase of DNA damage response (DDR) marker, such as 53BP1 foci formation27, throughout this study.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [28] Noda, A., Ning, Y., Venable, S. F., Pereira-Smith, O. M. & Smith, J. R. Cloning of senescent cell-derived inhibitors of DNA synthesis using an expression screen. Exp. Cell Res. 211, 90–98 (1994). source

      Not checked in the free run.

    • cites [29] Freund, A., Laberge, R. M., Demaria, M. & Campisi, J. Lamin B1 loss is a senescence-associated biomarker. Mol. Biol. Cell 23, 2066–2075 (2012). source

      Not checked in the free run.

    • cites [30] Shimi, T. et al. The role of nuclear lamin B1 in cell proliferation and senescence. Genes Dev. 25, 2579–2593 (2011). source

      Not checked in the free run.

    • cites [27] Imai, Y. et al. Crosstalk between the Rb pathway and AKT signaling forms a quiescence-senescence switch. Cell Rep. 7, 194–207 (2014). source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    BRD4 can bind 53BP1 and recruit it to DNA double-strand break sites, thereby facilitating NHEJ in a transcriptionally independent manner.

    “BRD4 has the potential to bind 53BP1, a DNA damage response protein serving as an adaptor for the assembly and activation of the DNA repair machinery, and recruit it to DSB sites, thereby facilitating NHEJ in a transcriptionally independent manner32,33.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [32] Stanlie, A., Yousif, A. S., Akiyama, H., Honjo, T. & Begum, N. A. Chromatin reader Brd4 functions in Ig class switching as a repair complex adaptor of nonhomologous end-joining. Mol. Cell 55, 97–110 (2014). source

      Not checked in the free run.

    • cites [33] Li, X. et al. BRD4 promotes DNA repair and mediates the formation of TMPRSS2-ERG gene rearrangements in prostate cancer. Cell Rep. 22, 796–808 (2018). source

      Not checked in the free run.

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

    Study design and conductnone given

    All experiments except Fig. 1a were repeated at least twice independently with similar results.

    “All experiments except Fig. 1a was repeated at least twice independently with similar results.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Senescent cells are viable for long periods and accumulate with age in various organs and tissues.

    “senescent cells are viable for long periods of time and thereby accumulate with age in various organs and tissues5–7.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [5] Krishnamurthy, J. et al. Ink4a/Arf expression is a biomarker of aging. J. Clin. Invest. 114, 1299–1307 (2004). source

      Not checked in the free run.

    • cites [6] Herbig, U., Ferreira, M., Condel, L., Carey, D. & Sedivy, J. M. Cellular senescence in aging primates. Science 311, 1257 (2006). source

      Not checked in the free run.

    • cites [7] Yamakoshi, K. et al. Real-time in vivo imaging of p16Ink4a reveals cross talk with p53. J. Cell Biol. 186, 393–407 (2009). source

      Not checked in the free run.

  • Classified as probably not a substantial claim.

    Premisecitation

    BRD4 inhibition induces autophagic gene expression.

    “BRD4 inhibition has recently been reported to induce autophagic gene expression22.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [22] Sakamaki, J. I. et al. Bromodomain protein BRD4 is a transcriptional repressor of autophagy and lysosomal function. Mol. Cell 66, 517–532.e519 (2017). source

      Not checked in the free run.

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

    Interpretationreasoning

    The senolytic effects of BETd have not been shown to be common to all types of senescent cells in vivo.

    “it remains unclear whether the senolytic effects of BETd are common to all types of senescent cells in vivo.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Current knowledge of senescent cells is largely based on cell culture systems, and efforts to characterise senescent cells in vivo have been limited due to the lack of absolute markers for senescent cells.

    “Since our current knowledge of senescent cells is largely based on cell culture systems, and efforts to characterise senescent cells in vivo have been limited due to the lack of absolute markers for senescent cells1”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [1] Gorgoulis, V. et al. Cellular senescence: defining a path forward. Cell 179, 813–827 (2019). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    BETd is a promising senolytic drug, identified through an unbiased high-throughput screening of around 47,000 small molecules.

    “In the present study, we have identified BETd as a promising senolytic drug, through an unbiased high-throughput screening of chemical compound libraries consisting of around 47,000 small molecules.”
    • Also stated in: § Discussion · hedged
      “further support the idea that BETd could be a promising senolytic drug.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    XRCC4 forms a complex with DNA ligase IV and plays an important role in non-homologous end joining repair for DNA double-strand breaks.

    “The protein encoded by the xrcc4 gene (XRCC4) forms a complex with DNA ligase IV (LIG4) and plays an important role in non-homologous end joining (NHEJ) repair for DNA double-strand breaks (DSBs)31.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [31] Polo, S. E. & Jackson, S. P. Dynamics of DNA damage response proteins at DNA breaks: a focus on protein modifications. Genes Dev. 25, 409–433 (2011). source

      Not checked in the free run.

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

    Study design and conductnone given

    The tissue culture cells were confirmed to be free of mycoplasma contamination.

    “We have confirmed the absence of mycoplasma contamination in our tissue culture cells.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    Aberrant activation of autophagy results in caspase-dependent apoptotic cell death.

    “the recent observation that the aberrant activation of autophagy results in caspase-dependent apoptotic cell death38”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [38] Chung, Y. et al. Dysregulated autophagy contributes to caspase-dependent neuronal apoptosis. Cell Death Dis. 9, 1189 (2018). source

      Not checked in the free run.

  • May combine two claims.

    Interpretationown data

    BETd treatment overrides the steady-state of NHEJ activation and autophagy gene repression, causing autophagy.

    “BETd treatment, however, over-rides this steady-state and thereby causing autophagy (right).”
    Quote in the paperSubstantial claim

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

    Interpretationreasoning

    All aspects of BRD4 inhibition-induced senolysis might not be explained by the factors described in the paper.

    “It should be noted, however, that all aspects of the BRD4 inhibition-induced senolysis might not be explained by the factors described here.”
    Quote in the paperNot a substantial claim
32 of 32

  • The paper relies on this without support.

    DSBs occur less frequently in quiescent cells.

    “However, since DSBs occur less frequently in quiescent cells, the blockade of NHEJ is likely to have a relatively minor impact in quiescent cells”
1 of 1

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?

  • A small molecule that degrades BRD4 has senolytic activity.

    “However, a recent chemical screen identified a small molecule that degrades BRD4 and has senolytic activity104.”

    From Cellular senescence in ageing: from mechanisms to therapeutic opportunities (2021) · cites this paper as [104]

    The Results section reports that ARV825 degrades BRD4 and has strong senolytic activity. However, the high-throughput screen identified BET inhibitors; ARV825 was tested afterward to confirm and extend the screening results, not identified as a screen hit. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • In mice with doxorubicin-induced senescence, the BET family protein degrader ARV-825 had effective senolytic activity against senescent hepatic stellate cells and delayed liver cancer development.

    “In mice with doxorubicin- induced senescence, the BET family protein degrader ARV-825 had effective senolytic activity against senescent hepatic stellate cells and delayed liver cancer development 206 .”

    From Senescence and cancer — role and therapeutic opportunities (2022) · cites this paper as [206]

    The Results (‘ARV825 provokes senolysis in vivo,’ Fig. 2) report fewer senescent hepatic stellate cells and reduced liver cancer development after ARV825 treatment, but in an obesity-induced HCC mouse model, not a doxorubicin-induced one. The doxorubicin experiments in the same section (Fig. 3) instead concern HCT116 colon-cancer cells and xenograft tumours. The claim combines findings from two different models. (LLM, conf=0.90)

    Experimental — accuracy not yet measured

  • BETd reactivates autophagy in senescent cells to execute autophagic cell death.

    “autophagy is suppressed in senescent cells and reactivated by BETd to execute autophagic cell death”

    From The role of cellular senescence and SASP in tumour microenvironment (2023) · cites this paper as [114]

  • BETd effectively suppressed liver cancer development in mice.

    “this study showed that BETd effectively suppressed liver cancer development in mice [114]”

    From The role of cellular senescence and SASP in tumour microenvironment (2023) · cites this paper as [114]

  • Neutralizing antibodies against key SASP components or their receptors, including IL-6, IL-1α, IL-1β, and TNF, have demonstrated senomorphic effects.

    “A number of neutralizing antibodies directed against key components of the SASP or their receptors, including IL-6, IL-1α, IL-1β and TNF, also have shown senomorphic properties104,240–248 (TABLE 1).”

    From Cellular senescence in ageing: from mechanisms to therapeutic opportunities (2021) · cites this paper as [104]

    The Discussion (page unavailable) mentions that BET inhibition may downregulate SASP factors, but does not report senomorphic effects of neutralizing antibodies. The Methods (page unavailable) list an IL-6 antibody for staining, not for neutralization; searches for neutralizing antibodies against IL-6, IL-1α, IL-1β, and TNF found no supporting passage. (LLM, conf=0.76)

    Experimental — accuracy not yet measured

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Authors
Wakita, M., Takahashi, A., Sano, O., Loo, T. M., Imai, Y., Narukawa, M., Iwata, H., Matsudaira, T., Kawamoto, S., Ohtani, N., Yoshimori, T., & Hara, E.
Affiliations
  • Research Institute for Microbial Diseases, The University of Osaka (JP)
  • Japanese Foundation For Cancer Research (JP)
  • Takeda (Japan) (JP)
  • Immunology Frontier Research Center, The University of Osaka (JP)
  • Osaka City University (JP)
  • The University of Osaka (JP)
Corresponding author
Hara, Eiji
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2020-04-22
Volume
11
Issue
1
Pages
1935-1935
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-020-15719-6
PMC
PMC7176673
OpenAlex
W3018442280
Semantic Scholar
8d288c9c73c0318686a20d8a31079c416b93b2c8
Cited by
206
References
54
Citations by year
2026: 16 · 2025: 36 · 2024: 35 · 2023: 30 · 2022: 42 · 2021: 34 · 2020: 12 · 2019: 1
FWCI
17.74
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • RePEc: Research Papers in Economics (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Japan Agency for Medical Research and Development (19gm501000h0003, 19cm0106401h0004)
Topics
  • Health Sciences › Medicine › Physiology › Telomeres, Telomerase, and Senescence
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Degradation and Inhibitors
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Aging › Genetics, Aging, and Longevity in Model Organisms
Keywords
senolysis, cellular senescence, non-homologous end joining, autophagy, senolytics, hepatic stellate cells, obesity, liver cancer, chemotherapy-induced senescence, tumor xenografts
MeSH terms
Bromodomain Containing Proteins, Animals, Antineoplastic Agents, Autophagy, Humans, Mice, Inbred C57BL, Mice, Nude, Neoplasms, Transcription Factors, Cellular Senescence, Cell Cycle Proteins, Cell Line, Tumor, Mice, DNA End-Joining Repair

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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