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The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2

Nature Medicine volume 13, pages 486–491 (2007)Cite this article

A Corrigendum to this article was published on 06 December 2011

An Addendum to this article was published on 06 December 2011

This article has been updated

Abstract

MicroRNAs (miRNAs) are endogenous noncoding RNAs, about 22 nucleotides in length, that mediate post-transcriptional gene silencing by annealing to inexactly complementary sequences in the 3′-untranslated regions of target mRNAs1,2,3. Our current understanding of the functions of miRNAs relies mainly on their tissue-specific or developmental stage-dependent expression and their evolutionary conservation, and therefore is primarily limited to their involvement in developmental regulation and oncogenesis2. Of more than 300 miRNAs that have been identified, miR-1 and miR-133 are considered to be muscle specific4,5,6. Here we show that miR-1 is overexpressed in individuals with coronary artery disease, and that when overexpressed in normal or infarcted rat hearts, it exacerbates arrhythmogenesis. Elimination of miR-1 by an antisense inhibitor in infarcted rat hearts relieved arrhythmogenesis. miR-1 overexpression slowed conduction and depolarized the cytoplasmic membrane by post-transcriptionally repressing KCNJ2 (which encodes the K+ channel subunit Kir2.1) and GJA1 (which encodes connexin 43), and this likely accounts at least in part for its arrhythmogenic potential. Thus, miR-1 may have important pathophysiological functions in the heart, and is a potential antiarrhythmic target.

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Figure 1: miR-1 is arrhythmogenic in ischemic and normal hearts.
Figure 2: miR-1 silences GJA1 and KCNJ2 by repressing their translation.
Figure 3: Effects of miR-1 on levels of Cx43 and Kir2.1.
Figure 4: Effects of GJA1 and KCNJ2 knockdown by specific siRNAs on arrhythmia.

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Change history

  • 06 December 2011

     In the version of this article initially published, lanes from the original blot shown in Figure 2b (NIZ and IZ samples, blotting for 55-kDa Kir2.1 and GAPDH) were rearranged in the published figure. The two lanes at the far right of the published blot were on the far left of the original blot, so that the sequence of the lanes in the original blot was as follows (from left to right): IZ (WT miR-1 + AMO-1), IZ (MT miR-1), NIZ, IZ, IZ (AMO-1), IZ (WT miR-1). We have added white space to indicate that the blot is not continuous in the HTML and PDF versions of the article.

  • 06 December 2011

     Editors' note: With regard to the above article, the editors wish to notify readers of Nature Medicine that the Montreal Heart Institute announced on 2 September 2011 that an investigation had found evidence of misconduct in publications from the laboratory of Zhiguo Wang at that institution. The committee in charge of this investigation recommended that several publications from Z. Wang's laboratory be retracted (some of which had already been retracted over the summer); in addition, his laboratory at the Montreal Heart Institute was closed. One of the papers investigated was the Nature Medicine paper listed above, for which the corresponding authors were Z. Wang (affiliated with the Harbin Medical University, China and the Montreal Heart Institute, Canada) and Baofeng Yang (affiliated with the State-Province Key Laboratories of Biomedicine-Pharmaceutics of China and the Institute of Cardiovascular Research, Harbin Medical University, China). The investigating committee's report stated that they did not find evidence of misconduct for the data in the Nature Medicine paper that was generated at the Montreal Heart Institute; namely, Figure 1a ("CAD human" data only) and Figure 2f,g. According to the report, the remaining data in the paper were not generated at the Montreal Heart Institute and were not investigated. In correspondence with Nature Medicine, both Z. Wang and Yang stand by the data in the Nature Medicine paper. However, Z. Wang noted that, for Figure 2b, lanes from the original blot had been rearranged in the published figure, for which we are publishing a Corrigendum in this issue.

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Acknowledgements

We thank X. Yang for technical support and M.-A. Lupien for handling human tissues. This work was supported in part by the Canadian Institute of Health Research and Fonds de la Recherche de l'Institut de Cardiologie de Montreal (to Z.W.) and by the National Nature Science Foundation of China (30430780) and the Foundation of National Department of Science and Technology of China (2004CCA06700) (to B.Y.). Z.W. is a senior research scholar of the Fonds de Recherche en Sante de Quebec.

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Author notes

  1. Baofeng Yang, Huixian Lin and Jiening Xiao: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Pharmacology (State-Province Key Laboratories of Biomedicine-Pharmaceutics of China), Harbin Medical University, Harbin, Heilongjiang, 150086, China

    Baofeng Yang, Yanjie Lu, Baoxin Li, Ying Zhang, Chaoqian Xu, Yunlong Bai, Huizhen Wang & Guohao Chen

  2. Institute of Cardiovascular Research, Harbin Medical University, Harbin, Heilongjiang, 150086, China

    Baofeng Yang, Huixian Lin, Jiening Xiao, Yanjie Lu, Xiaobin Luo & Zhiguo Wang

  3. Research Center, Montreal Heart Institute, 5000 Belanger East, Montreal, H1T 1C8, PQ, Canada

    Huixian Lin, Jiening Xiao, Xiaobin Luo, Huizhen Wang & Zhiguo Wang

  4. Department of Medicine, University of Montreal, Montreal, H3C 3J7, PQ, Canada

    Huixian Lin, Jiening Xiao, Xiaobin Luo & Zhiguo Wang

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Contributions

B.Y. and Z.W. supervised the project and wrote the manuscript; H.L. and J.X. designed the experiments and conducted the luciferase and real-time RT-PCR experiments; X.L., B.L. and H.W. performed a part of the luciferase and western blot analysis; Y.B. and C.X. conducted patch-clamp recordings; Y.L., Y.Z. and G.C. designed and conducted the animal studies.

Corresponding authors

Correspondence to Baofeng Yang or Zhiguo Wang.

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Supplementary information

Supplementary Fig. 1

The complementary sequences between miR-1 and its putative sites within the 3′UTRs of human (H), rat (R) and mouse (M) GJA1 and KCNJ2 mRNAs, predicted with computational and bioinformatics-based approach using TargetScan hosted by Wellcome Trust Sanger Institute11. (PDF 176 kb)

Supplementary Fig. 2

Comparisons of connexin43 (Cx43) and Kir2.1 protein levels between membrane samples isolated from individuals with healthy control hearts (HH) and from individuals suffered from coronary artery diseases (CAD), determined by Western blot analysis. (PDF 105 kb)

Supplementary Fig. 3

Examination of the specificity of the anti-miR-1 antisense inhibitor oligonucleotides (AMO-1) to target miR-1 and miR-1 actions on arrhythmias and the protein levels of Cx43 and Kir2.1, using a negative control AMO-1 (AMO-1 with ten mismatched nucleotides, Mis-AMO-1). (PDF 123 kb)

Supplementary Fig. 4

Verification for the lack of effect of miR-1 on KCNH2 (encoding HERG K+ channel) expression, serving as negative control experiments for GJA1 and KCNJ2, and verification for the effectiveness of miRNAs and AMO-1 used in our study. (PDF 160 kb)

Supplementary Fig. 5

Representative images showing successful uptake and distribution of exogenous miR-1 (50 μg in 100 μl) 12 h after multiple-site (∼10 sites) intramuscular injection into the myocardium. (PDF 1826 kb)

Supplementary Table 1

Characteristics of individuals from whom the human ventricular preparations were obtained for use in the present study (PDF 71 kb)

Supplementary Table 2

Changes of QRS duration and QT interval in rat hearts with intramuscular injection of various constructs (PDF 96 kb)

Supplementary Methods (PDF 169 kb)

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Yang, B., Lin, H., Xiao, J. et al. The muscle-specific microRNA miR-1 regulates cardiac arrhythmogenic potential by targeting GJA1 and KCNJ2. Nat Med 13, 486–491 (2007). https://doi.org/10.1038/nm1569

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