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. 2021 Feb 24;22(5):2245.
doi: 10.3390/ijms22052245.

Selective Cardiomyocyte Oxidative Stress Leads to Bystander Senescence of Cardiac Stromal Cells

Hélène Martini  1   2 , Lise Lefevre  1 , Sylvain Sayir  2 , Romain Itier  2 , Damien Maggiorani  1 , Marianne Dutaur  1 , Dimitri J Marsal  1 , Jérôme Roncalli  1   2 , Nathalie Pizzinat  1 , Daniel Cussac  1 , Angelo Parini  1   2 , Jeanne Mialet-Perez  1 , Victorine Douin-Echinard  1
Affiliations

Selective Cardiomyocyte Oxidative Stress Leads to Bystander Senescence of Cardiac Stromal Cells

Hélène Martini et al. Int J Mol Sci. .

Abstract

Accumulation of senescent cells in tissues during normal or accelerated aging has been shown to be detrimental and to favor the outcomes of age-related diseases such as heart failure (HF). We have previously shown that oxidative stress dependent on monoamine oxidase A (MAOA) activity in cardiomyocytes promotes mitochondrial damage, the formation of telomere-associated foci, senescence markers, and triggers systolic cardiac dysfunction in a model of transgenic mice overexpressing MAOA in cardiomyocytes (Tg MAOA). However, the impact of cardiomyocyte oxidative stress on the cardiac microenvironment in vivo is still unclear. Our results showed that systolic cardiac dysfunction in Tg MAOA mice was strongly correlated with oxidative stress induced premature senescence of cardiac stromal cells favoring the recruitment of CCR2+ monocytes and the installation of cardiac inflammation. Understanding the interplay between oxidative stress induced premature senescence and accelerated cardiac dysfunction will help to define new molecular pathways at the crossroad between cardiac dysfunction and accelerated aging, which could contribute to the increased susceptibility of the elderly to HF.

Keywords: CCR2+ Macrophages; SASP; cardiac mesenchymal stromal cells; monoamine oxidase A; oxidative stress; stress-induced premature senescence.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
MAOA dependent oxidative stress in cardiomyocytes triggers stress induced premature senescence in cardiac stromal cells. (a,b) Representative images of SA-β Gal activity on heart cryosections of NTg and Tg MAOA mice (a) and quantification of the percentage of positive SA-β Gal cells per nuclei per field (b) (n = 3 mice per group). Scale bar, 50 µM. (c,d) Representative SA-β Gal staining (c, green) and percentage (d) of positive cardiac stromal cells from NTg (n = 8), NTg treated with 150µm H2O2 (n = 6) and Tg MAOA (n = 10) mice. DNA stained with DAPI (blue). Scale bar, 50 μm. (d,e) Representative immunostaining with anti-γH2AX antibody (e, green) and percentage (f) of positive cardiac stromal cells from NTg (n = 6) and Tg MAOA (n = 8) mice. DNA stained with DAPI (blue). White arrows indicated cells zoomed in. Scale bar, 50 μm. (g) Relative mRNA expression of CDKIs of cardiac stromal cells from Tg MAOA (n = 5) compared to NTg (n = 4) mice. (h) Correlation curve between percentage of positive SA-βGal cardiac stromal cells in vitro and fractional shortening (FS) of NTg (n = 4) and Tg MAOA (n = 5) mice. Data are expressed as means ± SEM. * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. NTg group.
Figure 2
Figure 2
Premature senescence of cardiac stromal cells is associated to expression of SASP factors. (a,b) mRNA expression (2e(−ΔCT)) relative to gapdh of cytokines and growth factors (a) and chemokines (b) of heart and stromal cells from Tg MAOA (n = 5) and NTg (n = 4). (c) Numbers of endothelial cells (ECs), immune cells (CD45) and cardiac mesenchymal stromal cells (cMSCs) per heart ventricles of NTg (n = 7) and Tg MAOA (n = 12) mice analyzed by flow cytometry. (d) Percentage of CD90 positive cMSCs of NTg (n = 7) and Tg MAOA (n = 12) mice determined by flow cytometry. (e) Numbers of CD31+ cMSCs per heart ventricles of NTg (n = 7) and Tg MAOA (n = 12) mice by flow cytometry. (f,g) Representative dot-plots of PDGFR-α and CD90 (f) and of Sca-1 and CD31 (g) expression by cMSCs. Percentages of cells positive for CD90 (f) or for CD31 (g) out of total cMSCs from NTg and Tg MAOA mice are shown. Data are expressed as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NTg group.
Figure 3
Figure 3
Oxidative stress induced premature senescence of cMSCs promotes CCR2-dependent monocyte recruitment. (a,b) Representative immunostaining of anti-Ki67 antibody (red; a) and percentage (b) of positive cMSCs isolated from NTg (n = 3) and Tg MAOA (n = 4) mice. DNA stained with DAPI (blue). Scale bar, 50 μm. (c) Relative mRNA expression of CDKIs from Tg MAOA cMSCs (n = 7) compared to NTg (n = 5). (d,e) mRNA expression of SASP factors (d) and of Ccl2 and Cx3cr1 chimiokines (e) by cMSCs and endothelial cells (ECs) isolated from NTg (n = 6) and Tg MAOA (n = 5) mice relative to gapdh gene expression. (f,g) Quantification and representative images of CFSE labelled monocytes pretreated or not with the CCR2 antagonist (RS 504393) at the lower part of the Fluoroblok insert after 2 h incubation with cMSCs from NTg (n = 3) or Tg MAOA mice (n = 4) in the bottom chamber compared to medium; nuclei: blue). Monocytes were pretreated (or not)). Nuclei were stained with DAPI (Blue), CFSE (green), DAPI (blue), pores: white. Scale bar, 50 μm. Data are expressed as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NTg group.
Figure 4
Figure 4
Cardiomyocyte oxidative stress triggers shift in macrophage population phenotype toward a mixed M1/M2 profile. (a) Numbers of CCR2+ and CCR2- macrophages in ventricles of NTg (n = 7) and Tg MAOA mice (n = 12). (b) Representative dot-plots of CCR2 and CD14 expression by cardiac macrophages of NTg and Tg MAOA mice determined by flow cytometry. Control staining of viable macrophages with isotypes is shown. (c,d) Percentage of cardiac macrophages positive for Ly6C and CD14 of NTg (n = 7) and Tg MAOA (n = 12) mice (c) and representative histograms (d) by flow cytometry. (e) Relative mRNA expression of chemokine receptors, Ccr2 and Cx3cr1 by cardiac macrophages isolated from Tg MAOA (n = 10) compared to NTg (n = 10) mice. (f,g) Relative mRNA expression of CDKIs by cardiac macrophages isolated from Tg MAOA (n = 7) compared to NTg (n = 6) mice (f) or from 20 month-old mice (n = 7–9) compared to young (n = 5–9) mice (g). (h,i) Relative mRNA expression of M2 markers (n = 5–7 mice per group) (h) and of M1/M2 cytokines (n = 9–10 mice per group) (i) by cardiac macrophages isolated from Tg MAOA compared to NTg mice. (j,k) Quantification (j) of protein expression of IL-1β relative to pro IL-1β by cardiac macrophages isolated from NTg (n = 7) and Tg MAOA (n = 10) mice and representative images (k) of cell lysate samples immunoblotted with anti IL-1β (upper panel) or with anti β-actin (lower panel) by capillary-based western blot (n = 3 mice per group). Data are expressed as means ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. NTg groupe or (f) vs. young group.
Figure 5
Figure 5
Cardiac macrophages of Tg MAOA mice have decreased expression of phagocytic receptors. (a) Relative mRNA expression of phagocytic receptors by cardiac macrophages of Tg MAOA compared to NTg mice (n = 10 mice per group). (b–e) Flow cytometry analysis of cardiac macrophages from NTg (n = 7) and Tg MAOA (n = 12) mice for M2 (b,c) and M1 (d,e) marker expression. Percentages of positive cells (b,d) and representative histograms (c,e) are shown. (e) Data are expressed as means ± SEM. ** p < 0.01, **** p < 0.0001 vs. NTg group.
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References

    1. Correia-Melo C., Marques F.D., Anderson R., Hewitt G., Hewitt R., Cole J., Carroll B.M., Miwa S., Birch J., Merz A., et al. Mitochondria are required for pro-ageing features of the senescent phenotype. EMBO J. 2016;35:724–742. doi: 10.15252/embj.201592862. - DOI - PMC - PubMed
    1. Moiseeva O., Bourdeau V., Roux A., Deschenes-Simard X., Ferbeyre G. Mitochondrial dysfunction contributes to oncogene-induced senescence. Mol. Cell. Biol. 2009;29:4495–4507. doi: 10.1128/MCB.01868-08. - DOI - PMC - PubMed
    1. Dai D.F., Santana L.F., Vermulst M., Tomazela D.M., Emond M.J., MacCoss M.J., Gollahon K., Martin G.M., Loeb L.A., Ladiges W.C., et al. Overexpression of catalase targeted to mitochondria attenuates murine cardiac aging. Circulation. 2009;119:2789–2797. doi: 10.1161/CIRCULATIONAHA.108.822403. - DOI - PMC - PubMed
    1. Steinberg S.F. Oxidative stress and sarcomeric proteins. Circ. Res. 2013;112:393–405. doi: 10.1161/CIRCRESAHA.111.300496. - DOI - PMC - PubMed
    1. Terman A., Kurz T., Navratil M., Arriaga E.A., Brunk U.T. Mitochondrial turnover and aging of long-lived postmitotic cells: The mitochondrial-lysosomal axis theory of aging. Antioxid. Redox. Signal. 2010;12:503–535. doi: 10.1089/ars.2009.2598. - DOI - PMC - PubMed

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