Mol Ther Nucleic Acids. 2020 Jan 23
Yue Qiu, Rongchao Cheng, Chaoqi Liang, Yuan Yao, Wenhao Zhang, Jie Zhang, Mingyu Zhang, Baiyan Li, Chaoqian Xu,and Rong Zhang
Products used in the paper Details Operation
AAV vector packaging rAAV was used as carrier, among which rAAV9 is the most efficient vector for myocardial transduction. For this regard, rAAV9-antimiR- 20b or rAAV9-NC was produced (PackGene Biotech, Guangzhou, China) and the sequences were delivered into mouse, respectively, through tail vein injection at 1 x 10^11 vg (viral genomes) per animal at 5 weeks before TAC. Request Quote

Research Field: heart

Dose: 1×10^11 vg (viral genomes) per animal

Routes of Administration: tail vein injection

Targeted organ: heart

Animal or cell line strain: Adult male Kunming mice (weight 22–25 g)


MicroRNA (miRNA) and mitofusin-2 (Mfn2) are important in the development of cardiac hypertrophy, but the target relationship and mechanism associated with Ca2+ handling between SR and mitochondria under hypertrophic condition is not established. Mfn2 expression, Mfn2-mediated interorganelle Ca2+ cross-talk, and target regulation by miRNA-20b (miR-20b) were evaluated using animal/cellular hypertrophic models with state-of-the-art techniques. The results demonstrated that Mfn2 was downregulated and miR-20b was upregulated upon the target binding profile under hypertrophic condition. Our data showed that miR-20b induced cardiac hypertrophy that was reversed by recombinant adeno-associated virus vector 9 (rAAV9)-anti-miR-20b or miR-20b antisense inhibitor (AMO-20b). The deleterious action of miR-20b on Mfn2 expression/function and mitochondrial ATP synthesis was observed and reversed by rAAV9-anti-miR-20b or AMO-20b. The targeted regulation of miR-20b on Mfn2 was confirmed by luciferase reporter and miRNA-masking. Importantly, the facts that mitochondrial calcium uniporter (MCU) activation by Spermine increased the cytosolic Ca2+ into mitochondria, manifested as enhanced histamine-mediated Ca2+ release from mitochondrial, suggesting that Ca2+ reuptake/buffering capability of mitochondria to cytosolic Ca2+ is injured by miR-20b-mediated Mfn2 signaling, by which leads cytosolic Ca2+ overload and cardiac hypertrophy through Ca2+ signaling pathway. In conclusion, pro-hypertonic miR-20b plays crucial roles in cardiac hypertrophy through downregulation of Mfn2 and cytosolic Ca2+ overload by weakening the buffering capability of mitochondria.

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