Department of Endocrinology, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, People's Republic of China.
Department of Gynaecology and Obstetrics, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, People's Republic of China.
Am J Physiol Endocrinol Metab. 2024 Sep 1;327(3):E328-E343. doi: 10.1152/ajpendo.00123.2024. Epub 2024 Jul 24.
One of the sensitive markers for autoimmune thyroid disease (AITD) clinical identification is thyroid-stimulating hormone receptor antibodies (TRAbs). To quickly distinguish TRAb with distinct antigenic epitopes, a straightforward and uncomplicated technique has not yet been created. The objective of this study is to search for molecular diagnostic targets for different types of AITD {Graves' disease (GD), Graves' orbitopathy (GO), GD with third-degree goiter [GD(3)], hypothyroidism combined with positive TRAb [HT(TRAb+)]} as molecular diagnostic targets. Following action on thyroid cells, differential genes (DEGs) generated by TRAb with distinct antigenic epitopes were detected and identified by RNA sequencing (RNA-Seq), bioinformatics analysis, and quantitative reverse transcription-polymerase chain reaction (RT-qPCR) in the serum of patients with AITD. Using the 5-ethynyl-2'-deoxyuridine (EdU) assay, the effect of coculturing thyroid cells with different antigenic TRAb epitopes on the cells' capacity to proliferate was investigated. Bioinformatics analysis and RT-qPCR validation identified one GD key gene alpha 2-HS glycoprotein (), two GO key genes [adrenoceptor alpha 1D () and H2B clustered histone 18 ()], two GD(3) key genes [suppressor of cytokine signaling 1 ( and cytochrome b-245 beta ()], and one HT(TRAb+) key gene (MASP2). Correlation analysis and ROC curves showed that the abovementioned genes could be used as molecular diagnostic targets for different types of AITD. Finally, EdU results showed that TRAb inhibited thyroid cell proliferation in the HT(TRAb+) group compared with the normal control group, whereas the remaining three groups promoted thyroid cell proliferation, with a statistically significant difference ( < 0.05). We identified six key genes for different types of AITD, which have diagnostic value for different types of AITD. Meanwhile, we found that TRAbs with different antigenic epitopes in AITD have different biological functions. We identified six molecular targets of different types of AITD [GD, GO, GD(3), and HT(TRAb+)], which have diagnostic value for different types of AITD. Meanwhile, we found that TRAb with different antigenic epitopes extracted from the sera of patients with AITD had different biological functions, which also provided a new idea for further research on the mechanism of action of TRAb with different antigenic epitopes in AITD.
自身免疫性甲状腺疾病（AITD）临床识别的一个敏感标志物是促甲状腺激素受体抗体（TRAb）。为了快速区分具有不同抗原表位的 TRAb，目前还没有一种简单直接的方法。本研究旨在寻找不同类型 AITD（Graves 病（GD）、Graves 眼病（GO）、GD 伴三度甲状腺肿大[GD（3）]、伴阳性 TRAb 的甲状腺功能减退症[HT（TRAb+）]）的分子诊断靶标。通过对 TRAb 作用于甲状腺细胞后产生的差异基因（DEGs）进行检测，采用 RNA 测序（RNA-Seq）、生物信息学分析和定量逆转录聚合酶链反应（RT-qPCR）对 AITD 患者血清中的差异基因进行检测和鉴定。采用 5-乙炔基-2'-脱氧尿苷（EdU）检测法，研究了不同抗原性 TRAb 表位与甲状腺细胞共培养对细胞增殖能力的影响。生物信息学分析和 RT-qPCR 验证鉴定了 GD 的一个关键基因 alpha 2-HS 糖蛋白（）、GO 的两个关键基因[肾上腺素能受体 alpha 1D（）和 H2B 聚类组蛋白 18（）]、GD（3）的两个关键基因[细胞因子信号转导抑制因子 1（和细胞色素 b-245 beta（）]和 HT（TRAb+）的一个关键基因（甘露聚糖结合凝集素 2 相关丝氨酸蛋白酶 2（）。相关性分析和 ROC 曲线表明，上述基因可作为不同类型 AITD 的分子诊断靶标。最后，EdU 结果表明，与正常对照组相比，HT（TRAb+）组 TRAb 抑制了甲状腺细胞的增殖，而其余三组则促进了甲状腺细胞的增殖，差异有统计学意义（<0.05）。我们鉴定了六种不同类型 AITD 的关键基因，它们对不同类型 AITD 具有诊断价值。同时，我们发现 AITD 中不同抗原表位的 TRAb 具有不同的生物学功能。我们鉴定了六种不同类型 AITD 的分子靶标[GD、GO、GD（3）和 HT（TRAb+）]，它们对不同类型 AITD 具有诊断价值。同时，我们发现从 AITD 患者血清中提取的具有不同抗原表位的 TRAb 具有不同的生物学功能，这也为进一步研究 AITD 中不同抗原表位的 TRAb 作用机制提供了新的思路。
