Differentially expressed circular RNAs in orbital adipose/connective tissue from patients with thyroid-associated ophthalmopathy

Exp Eye Res. 2020 Jul:196:108036. doi: 10.1016/j.exer.2020.108036. Epub 2020 May 4.

Abstract

Our study aimed to investigate the differentially expressed circRNAs and their potential roles in orbital adipose/connective tissue from patients with thyroid-associated ophthalmopathy (TAO). The orbital adipose/connective tissue samples from three TAO patients and three control individuals were collected for RNA sequencing after depletion of ribosomal RNA. Differentially expressed mRNAs and up-regulated circRNAs were used for co-expression analysis. Functional and pathway enrichment analysis were conducted for the up- and down-regulated mRNAs in the circRNA-mRNA co-expression network. Meanwhile, circRNA-miRNA interaction network was established by miRanda software. The expression levels of mRNAs and circRNAs in control and TAO samples were determined by qRT-PCR. Among all the 16,329 circRNAs predicted from RNA sequencing data, 163 circRNAs (95 down-regulated and 68 up-regulated) were differentially expressed in TAO samples. Besides, 607 differentially expressed mRNAs were identified. The co-expression analysis showed circRNA_14940 was correlated with CCND1 and TNXB, while circRNA_10135 was correlated with PTGFR, and circRNA_14936 was correlated with TNFRSF19. The up-regulated CCND1 participated in Wnt signaling pathway. The down-regulated TNXB was involved in the ECM-receptor interaction, focal adhesion, and PI3K-Akt signaling pathway. PTGFR participated in neuroactive ligand-receptor interaction and calcium signaling pathway. TNFRSF19 was involved in cytokine-cytokine receptor interaction. In the interaction network, circRNA_14936 could interact with hsa-miR-10392-3p, and circRNA_12367 could interact with hsa-miR-1228-3p. Moreover, the expression changes of MMP2, TNXB, PTGFR, CCND1, and TNFRSF19, as well as circRNA_14936, circRNA_14940, and circRNA_12367 were validated by qRT-PCR. In conclusion, the differentially expressed circRNAs might participate in pathogenesis of TAO, and we speculated that circRNA_14940-CCND1-Wnt signaling pathway might be an important regulatory axis.

Keywords: Circular RNA; RNA sequencing; Thyroid-associated ophthalmopathy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / metabolism*
  • Computational Biology
  • Connective Tissue / metabolism*
  • Cyclin D1 / genetics
  • Gene Expression Profiling
  • Gene Expression Regulation / physiology*
  • Graves Ophthalmopathy / genetics*
  • Graves Ophthalmopathy / metabolism
  • Humans
  • Orbit / metabolism*
  • RNA, Circular / genetics*
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Receptors, Prostaglandin / genetics
  • Receptors, Tumor Necrosis Factor / genetics
  • Sequence Analysis, RNA
  • Signal Transduction
  • Tenascin / genetics
  • Up-Regulation

Substances

  • CCND1 protein, human
  • RNA, Circular
  • RNA, Messenger
  • Receptors, Prostaglandin
  • Receptors, Tumor Necrosis Factor
  • TNFRSF19 protein, human
  • Tenascin
  • prostaglandin F2alpha receptor
  • tenascin X
  • Cyclin D1