MiR-431 promotes cardiomyocyte proliferation by targeting FBXO32 expression

IF 3.2 4区医学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of Gene Medicine Pub Date : 2024-01-06 DOI:10.1002/jgm.3656

Mengsha Li, Chenrui Zhang, Lirong Tan, Tingyan Liu, Tingting Zhu, Xuejiao Wei, Jiacai Liu, Xiaoyun Si, Bing Li

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Abstract

Background

The induction of cardiomyocyte (CM) proliferation is a promising approach for cardiac regeneration following myocardial injury. MicroRNAs (miRNAs) have been reported to regulate CM proliferation. In particular, miR-431 expression decreases during cardiac development, according to Gene Expression Omnibus (GEO) microarray data. However, whether miR-431 regulates CM proliferation has not been thoroughly investigated.

Methods

We used integrated bioinformatics analysis of GEO datasets to identify the most significantly differentially expressed miRNAs. Real-time quantitative PCR and fluorescence in situ hybridization were performed to determine the miRNA expression patterns in hearts. Gain- and loss-of-function assays were conducted to detect the role of miRNA in CM proliferation. Additionally, we detected whether miR-431 affected CM proliferation in a myocardial infarction model. The TargetScan, miRDB and miRWalk online databases were used to predict the potential target genes of miRNAs. Luciferase reporter assays were used to study miRNA interactions with the targeting mRNA.

Results

First, we found a significant reduction in miR-431 levels during cardiac development. Then, by overexpression and inhibition of miR-431, we demonstrated that miR-431 promotes CM proliferation in vitro and in vivo, as determined by immunofluorescence assays of 5-ethynyl-2'-deoxyuridine (EdU), pH3, Aurora B and CM count, whereas miR-431 inhibition suppresses CM proliferation. Then, we found that miR-431 improved cardiac function post-myocardial infarction. In addition, we identified FBXO32 as a direct target gene of miR-431, with FBXO32 mRNA and protein expression being suppressed by miR-431. FBXO32 inhibited CM proliferation. Overexpression of FBXO32 blocks the enhanced effect of miR-431 on CM proliferation, suggesting that FBXO32 is a functional target of miR-431 during CM proliferation.

Conclusion

In summary, miR-431 promotes CM proliferation by targeting FBXO32, providing a potential molecular target for preventing myocardial injury.

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MiR-431 通过靶向 FBXO32 的表达促进心肌细胞增殖

背景诱导心肌细胞（CM）增殖是心肌损伤后心脏再生的一种可行方法。据报道，微小核糖核酸（miRNA）可调控心肌细胞的增殖。特别是，根据基因表达总库（GEO）的微阵列数据，miR-431 的表达在心脏发育过程中会减少。然而，miR-431 是否调控 CM 增殖尚未得到深入研究。方法我们对 GEO 数据集进行了综合生物信息学分析，以确定差异表达最显著的 miRNA。通过实时定量 PCR 和荧光原位杂交确定 miRNA 在心脏中的表达模式。我们还进行了功能增益和功能缺失试验，以检测 miRNA 在 CM 增殖中的作用。此外，我们还检测了 miR-431 是否会影响心肌梗死模型中 CM 的增殖。我们使用 TargetScan、miRDB 和 miRWalk 在线数据库预测 miRNA 的潜在靶基因。荧光素酶报告实验用于研究 miRNA 与靶标 mRNA 的相互作用。结果首先，我们发现在心脏发育过程中，miR-431 的水平明显下降。然后，通过过表达和抑制 miR-431，我们证明了 miR-431 在体外和体内促进了 CM 的增殖，这是由 5-乙炔基-2'-脱氧尿苷（EdU）、pH3、极光 B 和 CM 计数的免疫荧光测定确定的，而抑制 miR-431 则抑制了 CM 的增殖。随后，我们发现 miR-431 能改善心肌梗死后的心脏功能。此外，我们还发现 FBXO32 是 miR-431 的直接靶基因，miR-431 可抑制 FBXO32 mRNA 和蛋白的表达。FBXO32 可抑制 CM 增殖。过表达 FBXO32 可阻断 miR-431 对 CM 增殖的增强作用，这表明 FBXO32 是 miR-431 在 CM 增殖过程中的功能靶点。结论综上所述，miR-431 通过靶向 FBXO32 促进 CM 增殖，为预防心肌损伤提供了一个潜在的分子靶点。

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来源期刊

Journal of Gene Medicine 医学-生物工程与应用微生物

CiteScore

6.40

自引率

0.00%

发文量

审稿时长

6-12 weeks

期刊介绍： The aims and scope of The Journal of Gene Medicine include cutting-edge science of gene transfer and its applications in gene and cell therapy, genome editing with precision nucleases, epigenetic modifications of host genome by small molecules, siRNA, microRNA and other noncoding RNAs as therapeutic gene-modulating agents or targets, biomarkers for precision medicine, and gene-based prognostic/diagnostic studies. Key areas of interest are the design of novel synthetic and viral vectors, novel therapeutic nucleic acids such as mRNA, modified microRNAs and siRNAs, antagomirs, aptamers, antisense and exon-skipping agents, refined genome editing tools using nucleic acid /protein combinations, physically or biologically targeted delivery and gene modulation, ex vivo or in vivo pharmacological studies including animal models, and human clinical trials. Papers presenting research into the mechanisms underlying transfer and action of gene medicines, the application of the new technologies for stem cell modification or nucleic acid based vaccines, the identification of new genetic or epigenetic variations as biomarkers to direct precision medicine, and the preclinical/clinical development of gene/expression signatures indicative of diagnosis or predictive of prognosis are also encouraged.