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CARD9 promotes autophagy in cardiomyocytes in myocardial ischemia/reperfusion injury via interacting with Rubicon directly

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Abstract

Autophagy in cardiomyocyte is involved in myocardial ischemia/reperfusion (M-I/R) injury. Caspase recruitment domain-containing protein 9 (CARD9) plays a critical role in cardiovascular diseases (CVDs) such as hypertension and cardiac fibrosis. However, its role in autophagy following M-I/R injury is yet to be fully elucidated. Here, we found that CARD9 expression increased in M-I/R mouse hearts, and in H9c2 or neonatal rat ventricular myocytes (NRVMs) in response to hypoxia/reoxygenation (H/R) or H2O2. CARD9−/− mice exhibited a significant cardiac dysfunction following M-I/R injury (30 min of left ascending coronary (LAD) ischemia and 12 h of reperfusion) compared to wild-type (WT) mice. CARD9 deletion impaired autophagy during M-I/R in vivo and in vitro, evidenced by decrease of microtubule-associated protein 1 light chain 3 (LC3) lipidation and p62 accumulation. Conversely, CARD9 overexpression increased autophagic flux as indicated by enhanced expression of LC3 II/LC3 I and a reduction in p62. The protective effect of CARD9 on cardiomyocytes against H/R-induced oxidative stress was abolished by treatment with autophagy inhibitors, 3-methyladenine (3-MA) or Bafilomycin A1(BafA1). CARD9 interacted with RUN domain Beclin-1-interacting cysteine-rich-containing (Rubicon), a negative regulator of autophagy, and enhanced UV-irradiation-resistance-associated gene (UVRAG)-Beclin1-phosphatidylinositol 3-kinase catalytic subunit type 3 (PI3KC3) interaction and UVRAG-Vps16-mediated Rab7 activation to promote autophagosome formation, maturation, and endocytosis. Ablation of Rubicon by siRNA effectively prevented the detrimental effect of CARD9 knockdown on cardiomyocytes. These results suggest that CARD9 has protective effects on the myocardium against M-I/R injury by activating autophagy and restoring autophagic flux in vivo and in vitro.

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Acknowledgements

This work was supported by the grants from the National Natural Science Foundation of China (81170113; 81770306; 81471868; 81671895), and the Province Natural Science Foundation of Hunan (2018JJ2547).

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  1. Yuanbin Li and Pengfei Liang contributed equally to the study.

Authors and Affiliations

  1. Department of Pathophysiology, Sepsis Translational Medicine Key Lab of Hunan Province, Xiangya School of Medicine, Central South University, Changsha, 410000, Hunan, China

    Yuanbin Li, Bimei Jiang, Yuting Tang, Meidong Liu, Hui Sun, Cheng Chen & Xianzhong Xiao

  2. Department of Burns and Plastic Surgery, Xiangya Hospital, Central South University, Changsha, 410000, Hunan, China

    Pengfei Liang

  3. Department of Medicine, Division of Cardiovascular Medicine, Center for Precision Medicine, University of Missouri School of Medicine, 1 Hospital Drive, CE344, Columbia, MO, 65212, USA

    Xuanyou Liu, Hong Hao & Zhenguo Liu

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  1. Yuanbin Li
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Li, Y., Liang, P., Jiang, B. et al. CARD9 promotes autophagy in cardiomyocytes in myocardial ischemia/reperfusion injury via interacting with Rubicon directly. Basic Res Cardiol 115, 29 (2020). https://doi.org/10.1007/s00395-020-0790-6

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