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

The DOI matches its Crossref record; 53 of the 54 references it cites were found in a registry or an index; none of the 38 sources its claims cite is checked for support yet.

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, Kawamoto, Shimpei, Ohtani, Naoko, Yoshimori, Tamotsu, Hara, Eiji·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.

Overview

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.

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Integrity signals

Registry

Crossref · 10.1038/s41467-020-15719-6

Cites

53 of 53 references verified0 citation errors · 0 not checkable

Statistics

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Figures

Fig. 1 HTS identified BET inhibitors as potent senolytic compounds. a Outline of the high-throughput screening (HTS) procedure for identifying senolytic compound. IMR90-ER:Ras cells44 were stimulated with or without 4OHT for 7 days to induce senescence. These cells were plated on 384-well plates and incubated for another 24 hrs, followed by treatment with Takeda chemical compound library consisting of 47, 000 small molecules for 3 days. These cells were then subjected to cell viability analysis using CellTiter-Glo Luminescent Cell Viability assay system (Promega), followed by apoptosis analysis using Caspase-Glo3/7 Assay system (Promega). b–e Early passage pre-senescent (control) normal human diploid fibroblasts (TIG-3 cells) were rendered senescent by serial passage (replicative senescence), treatment with 250 ng/ml doxorubicin for 10 days (+DXR) (b) or infection with retrovirus encoding oncogenic Ras (+HRasV12) (d). These senescent cells and control pre-senescent cells (control (b) and +Vector (d)) were then incubated with increasing concentration of ARV825 indicated at right for 4 days. Relative cell number was counted throughout the experiments and representative photographs of the cells in the indicated culture conditions are shown at the bottom of the b, d. For all graphs, error bars indicate mean ± s.d. (n = 3) (b, d). Cells treated with or without 10 nM ARV825 for 4 days were subjected to western blotting using the antibodies shown on the right (c, e). β-actin was used as a loading control. The representative data from three independent experiments were shown (b–e). Source data are provided as a Source Data file.
Fig. 1 HTS identified BET inhibitors as potent senolytic compounds. a Outline of the high-throughput screening (HTS) procedure for identifying senolytic compound. IMR90-ER:Ras cells44 were stimulated with or without 4OHT for 7 days to induce senescence. These cells were plated on 384-well plates and incubated for another 24 hrs, followed by treatment with Takeda chemical compound library consisting of 47, 000 small molecules for 3 days. These cells were then subjected to cell viability analysis using CellTiter-Glo Luminescent Cell Viability assay system (Promega), followed by apoptosis analysis using Caspase-Glo3/7 Assay system (Promega). b–e Early passage pre-senescent (control) normal human diploid fibroblasts (TIG-3 cells) were rendered senescent by serial passage (replicative senescence), treatment with 250 ng/ml doxorubicin for 10 days (+DXR) (b) or infection with retrovirus encoding oncogenic Ras (+HRasV12) (d). These senescent cells and control pre-senescent cells (control (b) and +Vector (d)) were then incubated with increasing concentration of ARV825 indicated at right for 4 days. Relative cell number was counted throughout the experiments and representative photographs of the cells in the indicated culture conditions are shown at the bottom of the b, d. For all graphs, error bars indicate mean ± s.d. (n = 3) (b, d). Cells treated with or without 10 nM ARV825 for 4 days were subjected to western blotting using the antibodies shown on the right (c, e). β-actin was used as a loading control. The representative data from three independent experiments were shown (b–e). Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 2 BETd reduces obesity-associated HCC development in mice on deleterious side effects of senescent cells in vivo. a Timeline of the experimental procedure13. Eut, euthanasia. Red arrow indicates a daily treatment with ARV825 (injected intraperitoneally with 5 mg/kg) or vehicle for 5 consecutive days. b Representative macroscopic photographs of livers. The average liver tumor numbers and their relative size distribution were shown right (HFD + vehicle, n = 20; HFD + ARV825, n = 16). c Immunofluorescence of liver section. HSCs were visualized by αSMA staining (green) and DNA was stained by 4′ 6,-diamidino-2-phenylindole (DAPI; blue)13. The histograms indicate the percentages of αSMA-expressing cells that were positive for indicated markers. The number of cells scored per group was as follows: n = 329 (p21 + ND), n = 355 (p21 + Vehicle), n = 351 (p21 + ARV825), n = 309 (Groα + ND), n = 286 (+Vehicle), n = 385 (+ARV825), n = 320 (IL-6), n = 460 (+Vehicle), n = 337 (+ARV825) over three biological independent animals. Error bars indicate mean ± standard deviation (s.d.). Statistical significance was determined with two-tailed unpaired Student’s t-test (b), one-way ANOVA followed by Holm-Sidak multiple comparison test (c), P values < 0.05 were considered significant. Source data are provided as a Source Data file.
Fig. 2 BETd reduces obesity-associated HCC development in mice on deleterious side effects of senescent cells in vivo. a Timeline of the experimental procedure13. Eut, euthanasia. Red arrow indicates a daily treatment with ARV825 (injected intraperitoneally with 5 mg/kg) or vehicle for 5 consecutive days. b Representative macroscopic photographs of livers. The average liver tumor numbers and their relative size distribution were shown right (HFD + vehicle, n = 20; HFD + ARV825, n = 16). c Immunofluorescence of liver section. HSCs were visualized by αSMA staining (green) and DNA was stained by 4′ 6,-diamidino-2-phenylindole (DAPI; blue)13. The histograms indicate the percentages of αSMA-expressing cells that were positive for indicated markers. The number of cells scored per group was as follows: n = 329 (p21 + ND), n = 355 (p21 + Vehicle), n = 351 (p21 + ARV825), n = 309 (Groα + ND), n = 286 (+Vehicle), n = 385 (+ARV825), n = 320 (IL-6), n = 460 (+Vehicle), n = 337 (+ARV825) over three biological independent animals. Error bars indicate mean ± standard deviation (s.d.). Statistical significance was determined with two-tailed unpaired Student’s t-test (b), one-way ANOVA followed by Holm-Sidak multiple comparison test (c), P values < 0.05 were considered significant. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 3 BETd increases the efficacy of chemotherapy against xenograft tumours in mice. a, b Control and senescent HCT116 induced by treatment with 200 ng/ml doxorubicin for 10 days (+DXR) were incubated with 10 nM ARV825 or vehicle for 4 days. Relative cell number was counted throughout the experiments and representative photographs of the cells in the indicated culture conditions are shown at the bottom of the a. Error bars indicate mean ± s.d. (n = 3) (a). Cells treated with 10 nM ARV825 (+) or vehicle (−) for 4 days were subjected to western blotting using the antibodies shown on the right (b). Representative data from three independent experiments were shown (a, b). c–e Timeline of the experimental procedure. Blue arrow indicates the daily treatment with DXR (injected intraperitoneally with 1 mg/kg) or vehicle for 7 consecutive days. Red arrow indicates a daily treatment with ARV825 (injected intraperitoneally with 5 mg/kg) or vehicle for 5 consecutive days (c). Tumor volume was calculated weekly from n = 4 (control + Vehicle), n = 5 (control + ARV825), n = 5 (+DXR + Vehicle), n = 4 (+DXR + ARV825) biologically independent animals using calipers (d). Error bars indicate mean± s.e.m (d). Serial sections of the biopsy samples of xenografted tumours were subjected to hematoxylin and eosin (HE) staining, immunohistochemistry for senescence markers (p21Waf1/Cip1/Sdi1 and 53BP1), DNA damage marker (γH2AX), autophagy marker (LC3B) and apoptosis marker (Cleaved caspase-3) (e). The histogram shown at the right of the panel indicates the percentages of positively stained areas (e). The number of positive signals in the images was calculated from n = 6 (p21 and 53BP1) or n = 10 (γH2AX, LC3B, and Cleaved caspase-3) biologically independent tissue sections over four biological independent animals. Error bars indicate mean ± s.d. (e). Statistical significance was determined with a Kruskal–Wallis followed by Dunn’s multiple comparison test (d), or one-way ANOVA followed by Tukey multiple comparison test (e). P values < 0.05 were considered significant. Source data are provided as a Source Data file.
Fig. 3 BETd increases the efficacy of chemotherapy against xenograft tumours in mice. a, b Control and senescent HCT116 induced by treatment with 200 ng/ml doxorubicin for 10 days (+DXR) were incubated with 10 nM ARV825 or vehicle for 4 days. Relative cell number was counted throughout the experiments and representative photographs of the cells in the indicated culture conditions are shown at the bottom of the a. Error bars indicate mean ± s.d. (n = 3) (a). Cells treated with 10 nM ARV825 (+) or vehicle (−) for 4 days were subjected to western blotting using the antibodies shown on the right (b). Representative data from three independent experiments were shown (a, b). c–e Timeline of the experimental procedure. Blue arrow indicates the daily treatment with DXR (injected intraperitoneally with 1 mg/kg) or vehicle for 7 consecutive days. Red arrow indicates a daily treatment with ARV825 (injected intraperitoneally with 5 mg/kg) or vehicle for 5 consecutive days (c). Tumor volume was calculated weekly from n = 4 (control + Vehicle), n = 5 (control + ARV825), n = 5 (+DXR + Vehicle), n = 4 (+DXR + ARV825) biologically independent animals using calipers (d). Error bars indicate mean± s.e.m (d). Serial sections of the biopsy samples of xenografted tumours were subjected to hematoxylin and eosin (HE) staining, immunohistochemistry for senescence markers (p21Waf1/Cip1/Sdi1 and 53BP1), DNA damage marker (γH2AX), autophagy marker (LC3B) and apoptosis marker (Cleaved caspase-3) (e). The histogram shown at the right of the panel indicates the percentages of positively stained areas (e). The number of positive signals in the images was calculated from n = 6 (p21 and 53BP1) or n = 10 (γH2AX, LC3B, and Cleaved caspase-3) biologically independent tissue sections over four biological independent animals. Error bars indicate mean ± s.d. (e). Statistical significance was determined with a Kruskal–Wallis followed by Dunn’s multiple comparison test (d), or one-way ANOVA followed by Tukey multiple comparison test (e). P values < 0.05 were considered significant. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 4 BETd accelerates DSBs in senescent cells. a Heat map representation of the expression of genes (from RNA-seq experiments) upon treatment of DXR-induced senescent TIG-3 cells with 10 nM ARV825 or vehicle for 2 days. The heat map key indicates log2-fold changes from baseline. b, c Early passage (control) TIG-3 cells were rendered senescent by serial passage (replicative senescence) or treatment with 250 ng/ml doxorubicin for 10 days (+DXR). These cells and control cells were treated with 10 nM ARV825 (+) or vehicle (−) for 4 days and were then subjected to immunofluorescence staining using the antibodies shown on the left (b) or to neutral comet assay (c). The number of γH2AX or 53BP1 foci, above threshold intensity per nucleus (n = 50), was quantified and shown at the bottom of the b. The average tail olive moments were shown at the bottom of the c. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (b, c). Error bars indicate mean ± s.d. The representative data from three independent experiments were shown (b, c). Statistical significance was determined with one-way ANOVA followed by Holm-Sidak multiple comparison test (b, c). P values < 0.05 were considered significant. Source data are provided as a Source Data file.
Fig. 4 BETd accelerates DSBs in senescent cells. a Heat map representation of the expression of genes (from RNA-seq experiments) upon treatment of DXR-induced senescent TIG-3 cells with 10 nM ARV825 or vehicle for 2 days. The heat map key indicates log2-fold changes from baseline. b, c Early passage (control) TIG-3 cells were rendered senescent by serial passage (replicative senescence) or treatment with 250 ng/ml doxorubicin for 10 days (+DXR). These cells and control cells were treated with 10 nM ARV825 (+) or vehicle (−) for 4 days and were then subjected to immunofluorescence staining using the antibodies shown on the left (b) or to neutral comet assay (c). The number of γH2AX or 53BP1 foci, above threshold intensity per nucleus (n = 50), was quantified and shown at the bottom of the b. The average tail olive moments were shown at the bottom of the c. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (b, c). Error bars indicate mean ± s.d. The representative data from three independent experiments were shown (b, c). Statistical significance was determined with one-way ANOVA followed by Holm-Sidak multiple comparison test (b, c). P values < 0.05 were considered significant. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 5 BETd blocks 53BP1 function in senescent cells. a Early passage (control) or late passage (replicative senescence) TIG-3 cells were treated with 10 nM ARV825 (+) or vehicle (−) for 4 days. These cells were then subjected to western blotting using the antibodies shown on the right after immunoprecipitation with the antibodies shown at the top of the panel (IP:). b–f Early passage (control) or late passage (replicative senescence) TIG-3 cells were transfected with previously validated two different siRNA oligos against 53BP1 or control siRNA oligo twice at 2 days intervals. These cells were then subjected to RT-qPCR analysis (b), western blotting analysis (c), the cell proliferation analysis (d), immunofluorescence staining using the antibodies shown on the left (e) or neutral comet assay (f). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the bottom of the panel indicates the relative cell number (d). The number of γH2AX foci, above threshold intensity per nucleus (n = 50) was quantified and shown at the bottom of the e. The average tail olive moments were shown at the bottom of the f. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (e, f). For all graphs, error bars indicate mean ± s.d. (n = 3 for b, n = 4 for d) and the representative data from three independent experiments were shown. Statistical significance was determined with two-tailed unpaired Student’s t-test (b), one-way ANOVA followed by Tukey multiple comparison test (d), one-way ANOVA followed by Holm-Sidak multiple comparison test (e, f). P values < 0.05 were considered significant. Source data are provided as a Source Data file.
Fig. 5 BETd blocks 53BP1 function in senescent cells. a Early passage (control) or late passage (replicative senescence) TIG-3 cells were treated with 10 nM ARV825 (+) or vehicle (−) for 4 days. These cells were then subjected to western blotting using the antibodies shown on the right after immunoprecipitation with the antibodies shown at the top of the panel (IP:). b–f Early passage (control) or late passage (replicative senescence) TIG-3 cells were transfected with previously validated two different siRNA oligos against 53BP1 or control siRNA oligo twice at 2 days intervals. These cells were then subjected to RT-qPCR analysis (b), western blotting analysis (c), the cell proliferation analysis (d), immunofluorescence staining using the antibodies shown on the left (e) or neutral comet assay (f). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the bottom of the panel indicates the relative cell number (d). The number of γH2AX foci, above threshold intensity per nucleus (n = 50) was quantified and shown at the bottom of the e. The average tail olive moments were shown at the bottom of the f. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (e, f). For all graphs, error bars indicate mean ± s.d. (n = 3 for b, n = 4 for d) and the representative data from three independent experiments were shown. Statistical significance was determined with two-tailed unpaired Student’s t-test (b), one-way ANOVA followed by Tukey multiple comparison test (d), one-way ANOVA followed by Holm-Sidak multiple comparison test (e, f). P values < 0.05 were considered significant. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 6 Autophagy activation is required for BETd-induced sebolysis. Early passage (control) or late passage (replicative senescence) TIG-3 cells were treated with or without chemicals (10 nM ARV825, 20 μM Nu7026, 6 μl/ml cationic lipid) as indicated at the top of the panel, for 4 days and were then subjected to the cell proliferation analysis (a), neutral comet assay (b) or to immunofluorescence staining using the antibody against LC3B (c). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the right of the panel indicates the relative cell number (a). The number of LC3B puncta above threshold intensity per cells (n = 50) was quantified and was shown at the right of the c. The average tail olive moments were shown at the bottom of the b. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (b). Error bars indicate mean ± s.d. (n = 4 for a). The representative data from three independent experiments were shown (a–c). Statistical significance was determined with one-way ANOVA followed by Tukey multiple comparison test (a), or one-way ANOVA followed by Holm-Sidak multiple comparison test (b–c). P values < 0.05 were considered significant. Source data are provided as a Source Data file.
Fig. 6 Autophagy activation is required for BETd-induced sebolysis. Early passage (control) or late passage (replicative senescence) TIG-3 cells were treated with or without chemicals (10 nM ARV825, 20 μM Nu7026, 6 μl/ml cationic lipid) as indicated at the top of the panel, for 4 days and were then subjected to the cell proliferation analysis (a), neutral comet assay (b) or to immunofluorescence staining using the antibody against LC3B (c). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the right of the panel indicates the relative cell number (a). The number of LC3B puncta above threshold intensity per cells (n = 50) was quantified and was shown at the right of the c. The average tail olive moments were shown at the bottom of the b. Each box indicates the median and 25% and 75% quantiles, and the whiskers represent the minimum and maximum observations (b). Error bars indicate mean ± s.d. (n = 4 for a). The representative data from three independent experiments were shown (a–c). Statistical significance was determined with one-way ANOVA followed by Tukey multiple comparison test (a), or one-way ANOVA followed by Holm-Sidak multiple comparison test (b–c). P values < 0.05 were considered significant. Source data are provided as a Source Data file.CC BY · PubMed Central
Fig. 7 BETd provokes senolysis via NHEJ inhibition and autophagy activation. a–c Early passage (control) or late passaged (replicative senescence) TIG3 cells were treated with or without chemicals (10 nM ARV825, 300 nM BafilomycinA1, 400 nM Q-VD-OHT), as indicated at the top of the panel, for 4 days and were then subjected to the cell proliferation analysis (a), immunofluorescence staining using the antibody against LC3B (b) or to western blotting using the antibodies shown on the right (c). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the right of the panel indicates the relative cell number (a). The number of LC3B puncta above threshold intensity per cells (n = 50) was quantified and was shown at the right of the b. Error bars indicate mean ± s.d (n = 4 for a) (a, b). The representative data from three independent experiments were shown (a to c). Statistical significance was determined with one-way ANOVA followed by Tukey multiple comparison test (a), or one-way ANOVA followed by Holm-Sidak multiple comparison test (b). P values < 0.05 were considered significant. (d), The model distinguishing mechanisms that may operate in the absence (left) or presence (right) of BETd in senescent cells. BRD4 protects senescent cells from autophagy-induced cell death by NHEJ activation and autophagy gene repression (left). BETd treatment, however, over-rides this steady-state and thereby causing autophagy (right). Source data are provided as a Source Data file.
Fig. 7 BETd provokes senolysis via NHEJ inhibition and autophagy activation. a–c Early passage (control) or late passaged (replicative senescence) TIG3 cells were treated with or without chemicals (10 nM ARV825, 300 nM BafilomycinA1, 400 nM Q-VD-OHT), as indicated at the top of the panel, for 4 days and were then subjected to the cell proliferation analysis (a), immunofluorescence staining using the antibody against LC3B (b) or to western blotting using the antibodies shown on the right (c). Representative photographs of the cells in the indicated culture conditions are shown and the histogram shown at the right of the panel indicates the relative cell number (a). The number of LC3B puncta above threshold intensity per cells (n = 50) was quantified and was shown at the right of the b. Error bars indicate mean ± s.d (n = 4 for a) (a, b). The representative data from three independent experiments were shown (a to c). Statistical significance was determined with one-way ANOVA followed by Tukey multiple comparison test (a), or one-way ANOVA followed by Holm-Sidak multiple comparison test (b). P values < 0.05 were considered significant. (d), The model distinguishing mechanisms that may operate in the absence (left) or presence (right) of BETd in senescent cells. BRD4 protects senescent cells from autophagy-induced cell death by NHEJ activation and autophagy gene repression (left). BETd treatment, however, over-rides this steady-state and thereby causing autophagy (right). Source data are provided as a Source Data file.CC BY · PubMed Central

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    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”
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Metadata

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
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
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › 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

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