Graves ophthalmopathy (GO), a manifestation of Graves’ disease, is an organ-specific autoimmune disease. Intravenous glucocorticoid therapy (ivGCs) is the first-line treatment for moderate-to-severe and active GO. However, ivGCs is only effective in 70%–80% of GO patients. Insensitive patients who choose 12-week ivGCs not only were delayed in treatment but also took the risk of adverse reactions of glucocorticoids. At present, there is still a lack of effective indicators to predict the therapeutic effect of ivGCs. Therefore, the purpose of this study is to find biomarkers that can determine the sensitivity of ivGCs before the formulation of treatment, and to clarify the mechanism of its regulation of ivGCs sensitivity. This study first characterized the miRNA profiles of plasma exosomes by miRNA sequencing to identify miRNAs differentially expressed between GO patients with significant improvement (SI) and non-significant improvement (NSI) after ivGCs treatment. Subsequently, we analyzed the function of the predicted target genes of differential miRNAs. According to the function of the target genes, we screened 10 differentially expressed miRNAs. An expanded cohort verification showed that compared with NSI patients, mir-885-3p was upregulated and mir-4474-3p and mir-615-3p were downregulated in the exosomes of SI patients. Based on statistical difference and miRNA function, mir-885-3p was selected for follow-up study. The in vitro functional analysis of exosomes mir-885-3p showed that exosomes from SI patients (SI-exo) could transfer mir-885-3p to orbital fibroblasts (OFs), upregulate the GRE luciferase reporter gene plasmid activity and the level of glucocorticoid receptor (GR), downregulate the level of inflammatory factors, and improve the glucocorticoid sensitivity of OFs. Moreover, these effects can be inhibited by the corresponding miR inhibitor. In addition, we found that high levels of mir-885-3p could inhibit the AKT/NFκB signaling pathway, upregulate the GRE plasmid activity and GR level, and downregulate the level of inflammatory factors of OFs. Moreover, the improvement of glucocorticoid sensitivity by mir-885-3p transmitted by SI-exo can also be inhibited by the AKT/NFκB agonist. Finally, through the in vivo experiment of the GO mouse model, we further determined the relationship between exosomes’ mir-885-3p sequence, AKT/NFκB signaling pathway, and glucocorticoid sensitivity. As a conclusion, plasma exosomes deliver mir-885-3p and inhibit the AKT/NFκB signaling pathway to improve the glucocorticoid sensitivity of OFs. Exosome mir-885-3p can be used as a biomarker to determine the sensitivity of ivGCs in GO patients.
Background:
Molecular alterations have been recognized as valuable diagnostic biomarkers for papillary thyroid carcinoma (PTC).
Objectives:
This study aimed to identify an immune-related gene signatures associated with PTC progression using a computational pipeline and to develop an expression-based panel for rapid PTC risk classification.
Methods:
RNA-seq data and clinical information for PTC samples were downloaded from The Cancer Genome Atlas, followed by an analysis of differentially expressed (DE) RNAs among high-risk PTC, low-risk PTC, and normal groups. Immune cell infiltration and protein–protein interaction analyses were performed to obtain DE RNAs related to immunity. Then, a competing endogenous RNA (ceRNA) network was constructed to identify hub genes for the construction of a diagnostic model, which was evaluated by a receiver operator characteristic curve. A manually curated independent sample cohort was constructed to validate the model
Results:
By analyzing the immune cell infiltration, we found that the infiltration of plasma cells and CD8+ T cells was more abundant in the high-risk groups, and 68 DE mRNAs were found to be significantly correlated with these immune cells. Then a ceRNA network containing 10 immune-related genes were established. The ten-gene panel (including DEPDC1B, ELF3, VWA1, CXCL12, SLC16A2, C1QC, IPCEF1, ITM2A, UST, and ST6GAL1) was used to construct a diagnostic model with specificity (66.3%), sensitivity (83.3%), and area under the curve (0.762) for PTC classification. DEPDC1B and SLC16A2 were experimentally validated to be differentially expressed between high-risk and low-risk patients.
Conclusion:
The 10 immune-related gene panel can be used to evaluate the risk of PTC during point-of-care testing with high specificity and sensitivity.
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