Whole-transcriptome analysis of rat cavernosum and identification of circRNA-miRNA-mRNA networks to investigate nerve injury erectile dysfunction pathogenesis

ABSTRACT

There is growing evidence that circular RNAs (circRNAs) play a vital role in many kinds of diseases, including erectile dysfunction (ED). Nevertheless, the role of circRNAs in cavernous nerve-damaging ED (CNI-ED) is unknown. Here, we aimed to discover novel circRNAs, probed their potential role in the CNI-ED, and construct a ceRNA network of circRNAs. Twelve male Sprague Dawley rats were randomly divided into 2 groups by us: bilateral cavernous nerve crush (BCNC) and control groups. Four weeks after surgery, the spongy smooth muscle tissue of the rat penis was sequenced using high-throughput full transcriptome sequencing. We analyzed the expression of circRNAs, miRNAs, and mRNAs in the two groups. Twenty circRNAs with significantly different expressions were selected for RT-qPCR. CeRNA network of circRNAs was established using Cytoscape. GO and KEGG analysis was done by R package. Sequencing showed that 4,587 circRNAs, 762 miRNAs, and 21,661 mRNAs were dysregulated in the BCNC group. The top 20 differentially expressed circRNAs were further verified via RT-qPCR. The ceRNA network contained ten circRNAs, six miRNAs, and 227 mRNAs, including 23 circRNA-miRNA pairs and 227 miRNA-mRNA pairs. GO and KEGG analysis suggested that these ten circRNAs could main regulate energy metabolism processes. A protein‐protein interaction network was constructed with the mRNAs in ceRNA network, and five hub genes were identified. Our study revealed a potential link between circRNAs, miRNAs, and mRNAs in CNI-ED, suggesting that circRNAs may contribute to the occurrence of ED by regulating the cellular energy metabolism in CNI-ED.

KEYWORDS:

• Erectile dysfunction
• circular RNA
• ceRNA network
• bioinformatics analysis
• energy metabolism

Research highlights:

1. The role of circRNAs in cavernous nerve-damaging ED was assessed.

2. A ceRNA network centered on circRNA about CNI-ED is constructed.

3. The occurrence of CNI-ED is related to the regulation of energy metabolism by circRNA.

4. We identified five hub gene in the ceRNA network that play important roles in CNI-ED.

Acknowledgements

The author would like to thank the members of the research group for their hard work and the guidance of Professor Bodong lv. Thank OmicStudio tools at https://www.omicstudio.cn/tool for Bioinformatic analysis.

Data Availability Statement

The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.

The sequencing results have been uploaded to the GEO database: GSE176099 study at:https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE176099

Ethics statement

Animal ethics approval for this study was approved by the Committee on the Ethics of Animal Experiments of Zhejiang Chinese Medical University.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Jie Huang and Jianxiong Ma drafted the manuscript. Bodong Lv and Qing Hu contributed to the design of the research. Jie Wang, Kang Zhou, Ke Ma contributed to the development of the statistical plan and statistical analysis of the data. Jiewen Huang and Fan Zhao contributed to the conception of the study. Jie Wang and Jianxiong Ma contributed to data management and prepared the retrospective data for analysis. All authors reviewed, read and approved the final manuscript.

Supplementary material

Supplemental data for this article can be accessed here.

Additional information

Funding

This work was supported by National Natural Science Foundation of China(No. 81804092,82004360,82174376,82074433), Zhejiang Provincial Natural Science Foundation of China(No.GF20H270004, LY19H270011) and Zhejiang Provincial Key Research Project of Traditional Chinese Medicine,(No.2018ZY007).