The long noncoding RNA THBS1-AS1 promotes cardiac fibroblast activation in cardiac fibrosis by regulating TGFBR1
Junteng Zhou, Geer Tian, Yue Quan, Qihang Kong, Fangyang Huang, Junli Li, Wenchao Wu, Yong Tang, Zhichao Zhou, Xiaojing Liu
Junteng Zhou, Geer Tian, Yue Quan, Qihang Kong, Fangyang Huang, Junli Li, Wenchao Wu, Yong Tang, Zhichao Zhou, Xiaojing Liu
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Research Article Cardiology

The long noncoding RNA THBS1-AS1 promotes cardiac fibroblast activation in cardiac fibrosis by regulating TGFBR1

Abstract

Cardiac fibrosis is associated with an adverse prognosis in cardiovascular disease that results in a decreased cardiac compliance and, ultimately, heart failure. Recent studies have identified the role of long noncoding RNA (lncRNA) in cardiac fibrosis. However, the functions of many lncRNAs in cardiac fibrosis remain to be characterized. Through a whole-transcriptome sequencing and bioinformatics analysis on a mouse model of pressure overload–induced cardiac fibrosis, we screened a key lncRNA termed thrombospondin 1 antisense 1 (THBS1-AS1), which was positively associated with cardiac fibrosis. In vitro functional studies demonstrated that the silencing of THBS1-AS1 ameliorated TGF-β1 effects on cardiac fibroblast (CF) activation, and the overexpression of THBS1-AS1 displayed the opposite effect. A mechanistic study revealed that THBS1-AS1 could sponge miR-221/222 to regulate the expression of TGFBR1. Moreover, under TGF-β1 stimulation, the forced expression of miR-221/222 or the knockdown TGFBR1 significantly reversed the THBS1-AS1 overexpression induced by further CF activation. In vivo, specific knockdown of THBS1-AS1 in activated CFs significantly alleviated transverse aorta constriction–induced (TAC-induced) cardiac fibrosis in mice. Finally, we demonstrated that the human THBS1-AS1 can also affect the activation of CFs by regulating TGFBR1. In conclusion, this study reveals that lncRNA THBS1-AS1 is a potentially novel regulator of cardiac fibrosis and may serve as a target for the treatment of cardiac fibrosis.

Authors

Junteng Zhou, Geer Tian, Yue Quan, Qihang Kong, Fangyang Huang, Junli Li, Wenchao Wu, Yong Tang, Zhichao Zhou, Xiaojing Liu

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Figure 8

The role of THBS1-AS1 on TGFBR1 in human activated cardiac fibroblasts.

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The role of THBS1-AS1 on TGFBR1 in human activated cardiac fibroblasts. ...
(A and B) GTEx bulk RNA-Seq data, expression of markers of cardiac fibrosis, and THBS1-AS1 in human hearts. The gene expression levels (TPM) are graphed normalized to their maxima. A 2-sided Wilcoxon rank-sum test was used. (C) The bulk RNA-Seq data of human THBS1-AS1 expression in heart tissue in healthy donors and patients with dilated cardiomyopathy (DCM) (GSE135055). The gene expression levels (TPM) are graphed normalized to their maxima. A 2-sided Wilcoxon rank-sum test was used. (D) The correlation between THBS1-AS1 and TGFBR1 analyzed by the Pearson’s correlation test (GSE152250). (E and F) The rank-based GO analysis (BP, CC, and MF) and the KEGG pathway enrichment analysis of genes significantly associated with THBS1-AS1 identifies the top 15 enriched pathways. (G) The gene expression of CTGF, POSTN, α-SMA, and THBS1-AS1 in human cardiac fibroblasts stimulated with TGF-β1 for 24 hours by qPCR (n = 6). An unpaired, 2-tailed t test was used. (H) The gene expression of TGFBR1 in cardiac fibroblasts after siRNA-mediated THBS1-AS1 knockdown by qPCR (n = 6). An unpaired, 2-tailed t test was used. (I) The gene expression of CTGF, α-SMA, and POSTN in cardiac fibroblasts after interfering with siRNA-mediated THBS1-AS1 knockdown alone or interfering with THBS1-AS1 knockdown and plasmid-mediated TGFBR1 overexpression at the same time with TGF-β1 stimulation (n = 6). One-way ANOVA, followed by a Bonferroni post hoc test, was used. *P < 0.05. DCM, dilated cardiomyopathy. GO, Gene Ontology; BP, biological process; MF, molecular function; CC, cellular component; KEGG, Kyoto Encyclopedia of Genes and Genomes.