RNA-Seq Profiling and Co-Expression Network Analysis of lncRNAs and mRNAs Reveal Novel Pathogenesis of Noise-Induced Hidden Hearing Loss

Abstract

Background: Long noncoding RNAs (lncRNAs) mediate essential epigenetic regulation in a wide range of biological processes and diseases. In this study, we aimed to uncover the novel pathogenesis in noise-induced hidden hearing loss (NIHHL). Methods: We developed murine NIHHL model and murine noise-induced hearing loss (NIHL), respectively. Auditory brainstem response (ABR) were used to detect hearing function. The ribbon synapses alteration was determined by immunofluorescence. We investigated the potential roles of lncRNAs in NIHHL model via second sequencing. LncRNA-mRNA co-expression network were generated to identify lncRNA-expression trend and potential lncRNA target genes. The screened potential target genes were verified by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Findings: We identified 71 lncRNAs and 290 mRNAs specifically dysregulated in NIHHL model. Further, an association of lncRNA-mRNA showed that 27 dysregulated lncRNA were successfully predicted an interaction with mRNAs. Finally, a further KEGG analysis showed that 6 signaling pathways were putative pathway in the process of NIHHL, and among them, adrenergic signaling is the most likely pathway. LncRNA Sept7 and mRNA GNAS were found to be able to differentiate NIHHL model from Control Group and NIHL model. Validation of up-regulated of mRNA GNAS and lncRNA Sept7 in NIHHL model compared to Control Group and NIHL model by qRT-PCR. Interpretation: lncRNA Sept7 and mRNA GNAS served a promoter in combination with adrenergic signaling pathway, which might reveal novel pathogenesis of NIHHL and also might be a group of potential biomarkers in the prognosis of NIHHL. Funding Statement: The research was generously funded by [the National Natural Science Foundation of China] under Grant [81830030]; [the National Natural Science Foundation of China] under Grant [81770997]; [the Beijing Science and Technology Commission - Education Commission Joint Fund Project] under Grant [KZ201810025040] and [the National Natural Science Foundation of China] under Grant [81771016]. Declaration of Interests: None of the authors have any relevant conflicts of interests to declare. Ethics Approval Statement: All the experiments and animal welfares were carried out in accordance with National Institute of Health Guide for Care and Use of Laboratory Animals which were revised in 1996. Besides, all the experiments were approved by the animal committee of the Shengjing Hospital of China Medical University (2019PS355K). And all the efforts were aimed to minimize the animals’ suffering and the number of sacrificing mice.