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Transcriptome Sequencing Reveals ceRNA Networks and Molecular Signatures in Myopic Mouse Retina
Xueting Wang1,2, Huiman Zhuang3,4, Yalong Dang5
1School of Information Engineering, Henan University of Science and Technology, Luoyang, China.
Purpose:
Long non-coding RNAs (lncRNAs) and circular RNAs (circRNAs) in the retina play crucial roles in myopia; however, their regulatory mechanisms remain unclear. This study aimed to investigate significant genes and related signaling pathways associated with myopia by constructing and analyzing competitive endogenous RNA (ceRNA) networks within the retina.
Materials And Methods:
We investigated the expression patterns of lncRNAs, circRNAs, microRNAs (miRNAs), and messenger RNAs (mRNAs) within the retina using a form-deprivation myopia mouse model to elucidate their regulatory mechanisms in myopia. Transcriptomic sequencing was performed on retinal cells obtained from a mouse myopia model, followed by differential expression and functional enrichment analyses. Relevant ceRNA networks (lncRNA-miRNA-mRNA and circRNA-miRNA-mRNA) were constructed. Key pathways in these networks were validated via quantitative real-time polymerase chain reaction and western blotting, while Immunohistochemistry and single-cell sequencing analyses were conducted to analyze significant gene distribution.
Results:
The model exhibited approximately -6D diopters after 14 days of form deprivation. Transcriptomic analysis identified 187 differentially expressed lncRNAs (DE lncRNAs), 22 DE circRNAs, 24 DE miRNAs, and 368 DE mRNAs. Enrichment analysis linked these differentially expressed genes to various retinal functions and pathways. Validation revealed that the TCONS_00102163-mmu-miR-540-3p-Kcnq2, TCONS_00127926-novel_234-Tepp, and novel_circ_0001750-mmu-miR-212-5p-Sstr3 pathways in the retina were involved in regulating myopia. All experiments were conducted in three independent biological replicates.
Abstract:
Conclusions: This study systematically elucidated the synergistic regulatory mechanisms of non-coding RNAs in the development of myopia by constructing a ceRNA regulatory network in the retina, and further validated key regulatory axes. This provides an important theoretical foundation for understanding the molecular mechanisms of myopia and developing novel intervention strategies.
