Identification of the molecular basis of doxorubicin-induced cardiotoxicity

Journal name:
Nature Medicine
Volume:
18,
Pages:
1639–1642
Year published:
DOI:
doi:10.1038/nm.2919
Received
Accepted
Published online

Doxorubicin is believed to cause dose-dependent cardiotoxicity through redox cycling and the generation of reactive oxygen species (ROS). Here we show that cardiomyocyte-specific deletion of Top2b (encoding topoisomerase-IIβ) protects cardiomyocytes from doxorubicin-induced DNA double-strand breaks and transcriptome changes that are responsible for defective mitochondrial biogenesis and ROS formation. Furthermore, cardiomyocyte-specific deletion of Top2b protects mice from the development of doxorubicin-induced progressive heart failure, suggesting that doxorubicin-induced cardiotoxicity is mediated by topoisomerase-IIβ in cardiomyocytes.

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Figures

  1. Time-dependent changes in the transcriptome after acute doxorubicin treatment.
    Figure 1: Time-dependent changes in the transcriptome after acute doxorubicin treatment.

    (a) IPA of changes in the transcriptome in cardiomyocytes isolated from mice treated with 25 mg doxorubicin (dox) per kilogram of body weight for 16 h. Control, vehicle only. (b) Representative images of DSBs in heart sections from mice with different Top2b genotypes. γ-H2AX staining is shown in red, and DAPI nuclear staining in blue. The total numbers of γ-H2AX–positive nuclei as a percentage of total number of nuclei is shown in the bar graph. n = 4 mice per group; scale bar, 50 μm. (c) Apoptosis in heart sections, as detected by TUNEL assay. TUNEL-positive nuclei (red) and the total number of nuclei were counted. The number of TUNEL-positive cells per heart section is shown in the bar graph. n = 4 mice per group. Scale bars, 200 μm. (d) IPA of changes in the transcriptome in cardiomyocytes isolated from mice treated with 25 mg doxorubicin per kilogram of body weight for 72 h. Control, vehicle control. (e) Quantitative RT-PCR validation of changes in the expression of selected genes involved in mitochondrial biogenesis and function. n = 3 mice per group. The mRNA expression levels of each gene were calculated by 2−ΔΔCt (comparative threshold cycle; see Online Methods). Expression was normalized to mouse Gapdh. (f) Western blot analysis of Ndufa3, Atp5a and Sdha in mitochondria isolated from cardiomyocytes of the indicated genotypes. Cox4i1 was used as a loading control. One-way analysis of variance was used to determine statistical significance. *P <0.05; **P <0.001; error bars, mean ± s.e.m. (b,c,e).

  2. Changes in mitochondrial function and structure after acute doxorubicin treatment, and in heart function after chronic doxorubicin treatment.
    Figure 2: Changes in mitochondrial function and structure after acute doxorubicin treatment, and in heart function after chronic doxorubicin treatment.

    (a) Mitochondrial membrane potential was assessed by JC-1 staining. Shown are representative images (×200) of JC-1 staining of cardiomyocytes isolated from mice of the indicated genotypes treated with doxorubicin (dox; 25 mg per kilogram of body weight for 72 h) or PBS (control). Scale bars, 100 μm. (b) Representative transmission electron micrographs of heart sections from mice treated with doxorubicin or PBS as in a. Scale bars, 1 μm. (ce) Left ventricular end systolic volume (LVESV) (c), end diastolic volume (LVEDV) (d) and ejection fraction (e) in mice before treatment (baseline), after tamoxifen but before doxorubicin treatment (post tamoxifen) and after chronic doxorubicin treatment (5 mg per kilogram of body weight, once a week for 5 weeks) (post doxorubicin). n = 7 mice per group (ce). Error bars, mean ± s.e.m. *P <0.01.

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Affiliations

  1. Department of Cardiology, The University of Texas MD Anderson Cancer Center, Houston, Texas, USA.

    • Sui Zhang,
    • Tasneem Bawa-Khalfe &
    • Edward T H Yeh
  2. Texas Heart Institute at St. Luke's Episcopal Hospital, Houston, Texas, USA.

    • Xiaobing Liu,
    • Long-Sheng Lu &
    • Edward T H Yeh
  3. Department of Vascular Surgery, Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.

    • Xiaobing Liu
  4. Department of Pharmacology, University of Medicine and Dentistry of New Jersey—Robert Wood Johnson Medical School, Piscataway, New Jersey, USA.

    • Yi Lisa Lyu &
    • Leroy F Liu

Contributions

E.T.H.Y. and L.F.L. conceived the project. S.Z., X.L., T.B.-K., L.-S.L. and Y.L.L. performed experiments and data analysis. E.T.H.Y. and S.Z. wrote the manuscript with editorial input from L.F.L. and Y.L.L.

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The authors declare no competing financial interests.

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