The long noncoding RNA CARDINAL attenuates cardiac hypertrophy by modulating protein translation
Xin He, Tiqun Yang, Yao Wei Lu, Gengze Wu, Gang Dai, Qing Ma, Mingming Zhang, Huimin Zhou, Tianxin Long, Youchen Yan, Zhuomin Liang, Chen Liu, William T. Pu, Yugang Dong, Jingsong Ou, Hong Chen, John D. Mably, Jiangui He, Da-Zhi Wang, Zhan-Peng Huang
Xin He, Tiqun Yang, Yao Wei Lu, Gengze Wu, Gang Dai, Qing Ma, Mingming Zhang, Huimin Zhou, Tianxin Long, Youchen Yan, Zhuomin Liang, Chen Liu, William T. Pu, Yugang Dong, Jingsong Ou, Hong Chen, John D. Mably, Jiangui He, Da-Zhi Wang, Zhan-Peng Huang
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Research Article Cardiology Development

The long noncoding RNA CARDINAL attenuates cardiac hypertrophy by modulating protein translation

Abstract

One of the features of pathological cardiac hypertrophy is enhanced translation and protein synthesis. Translational inhibition has been shown to be an effective means of treating cardiac hypertrophy, although system-wide side effects are common. Regulators of translation, such as cardiac-specific long noncoding RNAs (lncRNAs), could provide new, more targeted therapeutic approaches to inhibit cardiac hypertrophy. Therefore, we generated mice lacking a previously identified lncRNA named CARDINAL to examine its cardiac function. We demonstrate that CARDINAL is a cardiac-specific, ribosome-associated lncRNA and show that its expression was induced in the heart upon pathological cardiac hypertrophy and that its deletion in mice exacerbated stress-induced cardiac hypertrophy and augmented protein translation. In contrast, overexpression of CARDINAL attenuated cardiac hypertrophy in vivo and in vitro and suppressed hypertrophy-induced protein translation. Mechanistically, CARDINAL interacted with developmentally regulated GTP-binding protein 1 (DRG1) and blocked its interaction with DRG family regulatory protein 1 (DFRP1); as a result, DRG1 was downregulated, thereby modulating the rate of protein translation in the heart in response to stress. This study provides evidence for the therapeutic potential of targeting cardiac-specific lncRNAs to suppress disease-induced translational changes and to treat cardiac hypertrophy and heart failure.

Authors

Xin He, Tiqun Yang, Yao Wei Lu, Gengze Wu, Gang Dai, Qing Ma, Mingming Zhang, Huimin Zhou, Tianxin Long, Youchen Yan, Zhuomin Liang, Chen Liu, William T. Pu, Yugang Dong, Jingsong Ou, Hong Chen, John D. Mably, Jiangui He, Da-Zhi Wang, Zhan-Peng Huang

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Figure 6

RNA interactome reveals that CARDINAL interacts with the translational regulator DRG1.

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RNA interactome reveals that CARDINAL interacts with the translational r...
(A) Designs for 3 sets of RNA pull-downs. (B) Venn diagram showing the Cardinal-interacting proteins. (C) Relative enrichment of Cardinal, Cardinal-as, and Linc-p21 from HA-DRG1 and control IP (n = 3 for each group). Mass spec, mass spectrometry. (D) Western blot (WB) of HA-DRG1 in RNA pull-downs. HA, hemagglutinin. (E) Relative enrichment of Cardinal from IP in HL-1 cells. (F) Ribo-Seq coverages of hearts after sham surgery or 2 weeks after TAC surgery (PRJNA484227) over the Myh7 genomic locus (n = 3 for each group). Black arrows show a potential ribosome stalling site. (G) Western blot and (H) quantification of DRG1 and puromycin-incorporated protein in HL-1 cells 48 hours after RNA interference. Cells were treated with 1 μM puromycin for 30 minutes before harvesting (n = 3 for each group). (I) Western blot and (J) quantification of puromycin-incorporated protein in NRVCs 24 hours after stimulation. Cells were treated by 1 μM puromycin for 30 minutes before harvesting (n = 3 for each group). (K) Immunofluorescence staining and (L) cell size quantification of NRVCs 48 hours after stimulation (n ≥300 for each group). Scale bars: 50 μm. (M) RT-qPCR results of relative gene expression in NRVCs 24 hours after stimulation (n = 3 for each group). (I–M) NRVCs were treated with si-NC or si-Drg1 and stimulated by culture medium with or without PE (50 μM). (N) Proportion of proteins with 0–4 categories of stalling motif among upregulated proteins versus the remaining proteins. (O) Violin plots showing the number of stalling motifs among upregulated versus the remaining proteins. (P) Proportion of proteins with 0–4 kinds of stalling motif among proteins in “actin filament organization” (AFO) gene set versus the remaining proteins. (Q) Violin plots showing numbers of stalling motifs among proteins in AFO gene set versus the remaining proteins. **P < 0.01 and ***P < 0.001, by 2-tailed Student’s t test (C and H), Mann-Whitney U test (O and Q), or 2-way ANOVA with Tukey’s post hoc test (J, L, and M). NC, negative control.