EPAC1 inhibition protects the heart from doxorubicin-induced toxicity
- Université Paris-Saclay, Inserm, UMR-S 1180, France
- Institut des Maladies Metaboliques et Cardiovasculaires - I2MC, INSERM, Université de Toulouse, France
- Innovations Thérapeutiques en Hémostase - UMR-S 1140, INSERM, Faculté de Pharmacie, Université Paris Descartes, Sorbonne Paris Cité, France
- Basic Cardiac Research UCSD School of Medicine La Jolla, United States
- Faculté de Médecine, Université Paris-Saclay, France
- Inserm UMR_S 999, Hôpital Marie Lannelongue, France
- Laboratoire de Radio-Oncologie, CHUV, Switzerland
- Université Paris-Saclay, UVSQ, Inserm, France
Figures
Figure 1 with 1 supplement
Doxorubicin (Dox) induces DNA damages and activates mitochondrial pathway of apoptosis in cardiac myocytes.
(a–d) Cell death markers were recorded by flow cytometry in neonatal rat ventricular myocyte (NRVM) treated or not with Dox (1 µM) for 12 hr, 24 hr, 36 hr, and 48 hr. Results are presented as mean ± …
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Figure 1—source data 1
Raw data for Figure 1.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig1-data1-v3.zip
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Figure 1—source data 2
Uncropped WB for Figure 1.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig1-data2-v3.zip
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Figure 1—source data 3
Raw WB imaging for Figure 1.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig1-data3-v3.zip
Figure 1—figure supplement 1
Doxorubicin (Dox) induced apoptotic cell death in cardiac myocytes.
(a–c) Neonatal rat ventricular myocyte (NRVM) were left untreated or treated with Dox (1 µM) ± CE3F4 (10 µM) for 24 hr and analyzed by flow cytometry. Representative monoparametric histograms of (a) …
Figure 2
Doxorubicin (Dox) modulates cAMP-EPAC1 pathway in cardiac cells.
(a) Neonatal rat ventricular myocyte (NRVM) were untreated or treated with 1 µM Dox for 3 hr, 6 hr, 16 hr, and 24 hr and the level of EPAC1 protein was detected by western blot. Actin was used as a …
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Figure 2—source data 1
Raw data for Figure 2.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig2-data1-v3.zip
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Figure 2—source data 2
Uncropped WB for Figure 2.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig2-data2-v3.zip
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Figure 2—source data 3
Raw WB imaging for Figure 2.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig2-data3-v3.zip
Figure 3
Pharmacological inhibition of EPAC1, but not EPAC2, protects cardiomyocytes from doxorubicin (Dox)-induced DNA damage.
(a) Neonatal rat ventricular myocyte (NRVM) were untreated or treated with Dox (1 µM) ± the specific EPAC1 inhibitor CE3F4 (10 µM) for 16 hr and the level of the DNA damage marker H2AX-pS139 was …
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Figure 3—source data 1
Raw data for Figure 3.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig3-data1-v3.zip
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Figure 3—source data 2
Uncropped WB for Figure 3.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig3-data2-v3.zip
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Figure 3—source data 3
Raw WB imaging for Figure 3.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig3-data3-v3.zip
Figure 4
EPAC1 inhibition reduces doxorubicin (Dox)-induced mitochondrial apoptotic pathway.
(a, b) Neonatal rat ventricular myocyte (NRVM) were untreated or treated with Dox (1 µM) ± CE3F4 (10 µM) for 24 hr and analyzed by flow cytometry. Results are expressed as means ± SEM. **p<0.01, …
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Figure 4—source data 1
Raw data for Figure 4.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig4-data1-v3.zip
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Figure 4—source data 2
Uncropped WB for Figure 4.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig4-data2-v3.zip
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Figure 4—source data 3
Raw WB imaging for Figure 4.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig4-data3-v3.zip
Figure 5 with 1 supplement
Doxorubicin (Dox)-induced cardiotoxicity was prevented in EPAC1 knock-out (KO) mice.
WT (a–d) or EPAC1 KO mice (e–g) were injected (i.v.) three times with saline solution (Sal) or Dox at 4 mg/kg for each injection (12 mg/kg cumulative dose). Echocardiographic analysis was performed …
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Figure 5—source data 1
Raw data for Figure 5.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig5-data1-v3.zip
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Figure 5—source data 2
Uncropped WB for Figure 5.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig5-data2-v3.zip
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Figure 5—source data 3
Raw WB imaging for Figure 5.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig5-data3-v3.zip
Figure 5—figure supplement 1
Body weight of mice treated with doxorubicin (Dox).
WT and EPAC1 knock-out (KO) mice were injected (i.v.) three times with (a) saline solution (Sal) or (b) Dox at 4 mg/kg for each injection (12 mg/kg cumulative dose). The body weight was measured …
Figure 6
Doxorubicin (Dox)-induced cardiotoxicity was prevented in EPAC1 knock-out (KO) mice.
WT and EPAC1 KO mice were injected (i.v.) three times with saline solution (Sal) or Dox at 4 mg/kg for each injection (12 mg/kg cumulative dose). Ventricular cells were isolated from control and …
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Figure 6—source data 1
Raw data for Figure 6.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig6-data1-v3.zip
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Figure 6—source data 2
Uncropped WB for Figure 6.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig6-data2-v3.zip
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Figure 6—source data 3
Raw WB imaging for Figure 6.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig6-data3-v3.zip
Figure 7
EPAC1 inhibition enhances doxorubicin (Dox)-induced cytotoxicity in various human cancer cell lines.
Human MCF-7 breast cancer cells (a) and HeLa cervical cancer cells (b) were untreated or treated with the indicated doses of Dox ± CE3F4 (10 µM) for 24 hr. Cell death was measured by flow cytometry …
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Figure 7—source data 1
Raw data for Figure 7.
- https://cdn.elifesciences.org/articles/83831/elife-83831-fig7-data1-v3.zip
Author response image 1
