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  • RNA Profiling of TAO Orbital Connective Tissue

    By Lynda Seminara
    Selected by Prem S. Subramanian, MD, PhD
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    Journal Highlights

    Investigative Ophthalmology & Visual Science
    2022;63(12):27

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    Ye et al. looked at the molecular mechanisms of two clinical subtypes of thyroid-associated ophthalmopathy (TAO): type I (mainly fat) and type II (mainly muscle). Using high-through­put RNA sequencing (RNA-seq), they explored the expression profiles of circular RNAs (circRNAs). They found differences between the two subtypes and identified signaling pathways that may contribute to TAO pathogenesis and thus could be promising diagnostic and therapeutic targets.

    In this study, the high-throughput RNA-seq was performed on six pairs of type I and type II samples of orbital connective tissue from patients with TAO. In another three pairs of type I and type II samples, the expression levels of circRNA and messenger RNA (mRNA) were measured by quan­titative real-time polymerase chain reaction (qRT-PCR). Bioinformatics predictions were used to construct a circRNA-microRNA (miRNA) net­work. Subsequently, a protein-protein interaction network was constructed based on differential mRNA expression, and hub genes were determined from Cytoscape software. To determine circRNA function, pathway and functional enrichment analyses were performed. Western blotting was applied to evalu­ate lentiviral-mediated overexpression of hsa_circ_0007006 and the relation­ship between hsa_circ_0007006 and COL1A1 or MMP2. It also was used to identify differential pathways.

    Findings demonstrated that RNA-seq predicted 7,489 circRNAs and 15,803 mRNAs, which included 94 upregulated and 76 downregulated circRNAs and 488 upregulated and 138 downregulated mRNAs. The RNA-seq data were validated by qRT-PCR analysis of seven dysregulated circRNAs and two major mRNAs. The competing endogenous RNA network comprised seven circRNAs, 23 miRNAs, and 262 mRNAs. Functional analysis showed that overexpression of hsa_circ_0007006 led to decreased ex­pression of COL1A1 and MMP2. It also revealed differences in the activation level of the relaxin signaling pathway between the TAO subtypes, which was significantly higher in type II.

    According to the authors, the competing endogenous RNA analysis of potential target genes emphasizes the relationship between dysregulated circRNAs and the clinical subtype of TAO, which may provide a new perspective for research on TAO. The authors have begun exploring the in-depth mechanism and overall effect of hsa_circ_0007006 and relaxin signal­ing in TAO, which could be potential biomarkers and therapeutic targets for future studies, they said.

    The original article can be found here.