Hashimoto’s thyroiditis (HT) is the most common autoimmune disease characterized by lymphocytic infiltration and thyrocyte destruction. Dissection of the interaction between the thyroidal stromal microenvironment and the infiltrating immune cells might lead to a better understanding of HT pathogenesis. Here we show, using single-cell RNA-sequencing, that three thyroidal stromal cell subsets, ACKR1⁺endothelial cells and CCL21⁺myofibroblasts and CCL21⁺fibroblasts, contribute to the thyroidal tissue microenvironment in HT. These cell types occupy distinct histological locations within the thyroid gland. Our experiments suggest that they might facilitate lymphocyte trafficking from the blood to thyroid tissues, and T cell zone CCL21⁺fibroblasts may also promote the formation of tertiary lymphoid organs characteristic to HT. Our study also demonstrates the presence of inflammatory macrophages and dendritic cells expressing high levels of IL-1β in the thyroid, which may contribute to thyrocyte destruction in HT patients. Our findings thus provide a deeper insight into the cellular interactions that might prompt the pathogenesis of HT.

Objective: Congenital hypothyroidism (CH), the most common neonatal metabolic disorder, is characterized by impaired neurodevelopment. Although several candidate genes have been associated with CH, comprehensive screening of causative genes has been limited.Design and methods: One hundred ten patients with primary CH were recruited in this study. All exons and exon–intron boundaries of 21 candidate genes for CH were analyzed by next-generation sequencing. And the inheritance pattern of causative genes was analyzed by the study of family pedigrees.Results: Our results showed that 57 patients (51.82%) carried biallelic mutations (containing compound heterozygous mutations and homozygous mutations) in six genes (DUOX2,DUOXA2,DUOXA1,TG,TPOandTSHR) involved in thyroid hormone synthesis. Autosomal recessive inheritance of CH caused by mutations inDUOX2,DUOXA2,TGandTPOwas confirmed by analysis of 22 family pedigrees. Notably, eight mutations in four genes (FOXE1,NKX2-1,PAX8andHHEX) that lead to thyroid dysgenesis were identified in eight probands. These mutations were heterozygous in all cases and hypothyroidism was not observed in parents of these probands.Conclusions: Most cases of congenital hypothyroidism in China were caused by thyroid dyshormonogenesis rather than thyroid dysgenesis. This study identified previously reported causative genes for 57/110 Chinese patients and revealedDUOX2was the most frequently mutated gene in these patients. Our study expanded the mutation spectrum of CH in Chinese patients, which was significantly different from Western countries.

Graves' disease (GD), characterized by autoantibodies targeting antigens specifically expressed in thyroid tissues causing hyperthyroidism, is triggered by a combination of genetic and environmental factors. However, only a few loci for GD risk were confirmed in the various ethnic groups, and additional genetic determinants have to be detected. In this study, we carried out a three-stage study in 9529 patients with GD and 9984 controls to identify new risk loci for GD and found genome-wide significant associations in the overall populations for five novel susceptibility loci: the GPR174-ITM2A at Xq21.1, C1QTNF6-RAC2 at 22q12.3–13.1, SLAMF6 at 1q23.2, ABO at 9q34.2 and an intergenic region harboring two non-coding RNAs at 14q32.2 and one previous indefinite locus, TG at 8q24.22 (P_combined < 5 × 10⁻⁸). The genotypes of corresponding variants at 14q32.2 and 8q24.22 were correlated with the expression levels of C14orf64 and a TG transcript skipping exon 46, respectively. This study increased the number of GD loci with compelling evidence and indicated that non-coding RNAs might be potentially involved in the pathogenesis of GD.

Thyroid nodules (TNs) are highly prevalent worldwide and have a pattern of female predominance. Bisphenol A (BPA) is an endocrine disruptor that can lead to adverse effects in human health. However, epidemiologic studies revealing the association between BPA exposure and TNs are limited and the results are inconsistent. We aimed to examine the association between urinary BPA and TNs in women who are more susceptible to TNs. We conducted a case-control study with 1416 women aged 18 years or older (705 cases, 711 controls). All participants underwent thyroid ultrasonography. Urinary total BPA (free and conjugated) concentration was quantified using the HPLC-MS/MS. We analyzed the association between urinary BPA concentration and the risk of TNs using crude and multivariable logistic regression models. Participants were further stratified into thyroid autoantibody positive group (at least one positive) and thyroid autoantibody negative group (both negative) according to the thyroglobulin antibody (TGAb) and thyroid peroxidase antibody (TPOAb) levels, and restricted cubic spline regression was also applied to determine the possible nonlinear relationship between urinary BPA and TNs. Compared with women in the first quartile, the odds of TNs was 72% (adjusted OR = 1.72, 95% CI: 1.25 to 2.35) higher for those in the second quartile, 54% (adjusted OR = 1.54, 95% CI: 1.12 to 2.12) higher for those in the third quartile, and 108% (adjusted OR = 2.08, 95% CI: 1.50 to 2.90) higher for those in the fourth quartile after adjusting for age, BMI, education, HDL-C, LDL-C, triglyceride, total cholesterol, urinary iodine, TGAb and TPOAb. When the study population was stratified into thyroid autoantibody positive group and thyroid autoantibody negative group, we found that only in the positive group, the association was significant in model 1 (crude OR = 2.80; 95% CI = 1.90 to 4.12), model 2 (adjusted OR = 2.84; 95% CI = 1.91 to 4.22), model 3 (adjusted OR = 4.01; 95% CI = 2.57 to 6.27) and model 4 (adjusted OR = 3.71; 95% CI = 2.36 to 5.83). Multivariable-adjusted restricted cubic spline analysis demonstrated a similar result that in the thyroid autoantibody positive group, the association between urinary BPA and TNs risk was near linear (P-overall <0.001; P-non-linear = 0.054). In Chinese women, higher urinary BPA concentration was associated with increased risk of TNs only in those with positive thyroid autoantibodies. Moreover, this association was near linear, indicating that any rise in BPA exposure was associated with elevated TNs risk.

5-Methylcytosine (m5C) is one of the most ubiquitous modifications of mRNA and contributes to cancer pathogenesis. Aly/REF export factor (ALYREF), an m5C reader, is associated with the prognosis of liver hepatocellular carcinoma (LIHC). However, the effects of ALYREF on the progression of LIHC and the underlying molecular mechanisms remains elusive. Through an analysis of an online database and 3 independent LIHC cohorts, we found that ALYREF was markedly elevated in human liver cancer tissues and was significantly correlated with LIHC clinicopathological parameters, including Ki67⁺ cell rate, high-grade TNM stage, and poor prognosis. Several experiments were conducted to investigate the molecular basis and functional role of ALYREF-related progression in this study. ALYREF could enhance LIHC cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) in vitro and tumor formation in vivo. Mechanistically, ALYREF promoted the progression of human LIHC through EGFR pathways. Furthermore, ALYREF could directly bind to the m5C modification site of EGFR 3' untranslated region (3' UTR) to stabilize EGFR mRNA. Collectively, ALYREF played a crucial oncogenic role in LIHC via the stabilization of EGFR mRNA and subsequent activation of the STAT3 signaling pathway. Our results may help to elucidate the potential mechanisms of ALYREF-induced m5C modification in the progression of human LIHC.