N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance

Graves’ ophthalmopathy (GO) obvious manifestation is the imbalance of Th17/Treg. N6-methyladenosine (m6A) methylation is an important regulator of Th17/Treg balance. However, few reports narrate how m6A regulators mediate the role of genes in GO progression. We explored the m6A modification of THBS1...

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Main Authors: Lin-na Li, Jie-man Wu, Zong-ji Zheng, Shu-xian Li, Meng-yi Cai, Meng-chen Zou
Format: Article
Language:English
Published: Taylor & Francis Group 2025-12-01
Series:Autoimmunity
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Online Access:https://www.tandfonline.com/doi/10.1080/08916934.2024.2433628
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author Lin-na Li
Jie-man Wu
Zong-ji Zheng
Shu-xian Li
Meng-yi Cai
Meng-chen Zou
author_facet Lin-na Li
Jie-man Wu
Zong-ji Zheng
Shu-xian Li
Meng-yi Cai
Meng-chen Zou
author_sort Lin-na Li
collection DOAJ
description Graves’ ophthalmopathy (GO) obvious manifestation is the imbalance of Th17/Treg. N6-methyladenosine (m6A) methylation is an important regulator of Th17/Treg balance. However, few reports narrate how m6A regulators mediate the role of genes in GO progression. We explored the m6A modification of THBS1 mediated by WTAP, and the mechanism by which THBS1 regulated glycolysis and Th17/Treg balance. A total of 12 peripheral blood (4 GO samples, 4 GH samples, and 4 health samples) were collected to measure the percentage of Th17/Treg in monocytes by flow cytometry. RNA sequencing (RNA-seq) combined with MeRIP sequencing (MeRIP-seq) was used to screen differentially expressed and methylated genes. MeRIP-qPCR was performed to evaluate the m6A abundance of THBS1 after WTAP silencing. Glycolysis of CD4+ T cells was reflected by the lactate content and glucose uptake. The number of Th17 cells was increased in GO peripheral blood, whereas the Treg cells decreased. RNA-seq acquired 679 differentially expressed genes (308 up-regulated, and 371 down-regulated) in the CD4+ T cells of GO compared to healthy control. MeRIP-seq identified 3277 m6A peaks between the GO group and the healthy control group, corresponding with 2744 genes (1143 hypermethylated and 1601 hypomethylated). Combined analysis of RNA-seq and MeRIP-seq showed 81 hypermethylated and up-regulated genes. Among the six candidate genes in the PI3K-signaling pathway, THBS1 was the most significantly differentially expressed and hypermethylated. THBS1 silencing resulted in decreased lactate content and glucose uptake in CD4+ T cells. WTAP was significantly upregulated in CD4+ T cells of GO, and WTAP silencing significantly reduced m6A abundance and expression of THBS1. Upregulated and hypermethylated THBS1 mediated by WTAP promoted glycolysis of CD4+ T cells, affected Th17/Treg balance, and facilitated GO progression. We provided a novel potential target for GO treatment and revealed the molecular mechanism of WTAP and THBS1 in GO under the m6A perspective.
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issn 0891-6934
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language English
publishDate 2025-12-01
publisher Taylor & Francis Group
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series Autoimmunity
spelling doaj-art-a917d4b61f5e453186f6ed78407a93fb2024-12-17T21:57:41ZengTaylor & Francis GroupAutoimmunity0891-69341607-842X2025-12-0158110.1080/08916934.2024.2433628N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balanceLin-na Li0Jie-man Wu1Zong-ji Zheng2Shu-xian Li3Meng-yi Cai4Meng-chen Zou5Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, ChinaDepartment of Health Management, Nanfang Hospital Zengcheng Campus, Guangzhou, ChinaDepartment of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, ChinaDepartment of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, ChinaDepartment of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, ChinaDepartment of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, Guangdong, ChinaGraves’ ophthalmopathy (GO) obvious manifestation is the imbalance of Th17/Treg. N6-methyladenosine (m6A) methylation is an important regulator of Th17/Treg balance. However, few reports narrate how m6A regulators mediate the role of genes in GO progression. We explored the m6A modification of THBS1 mediated by WTAP, and the mechanism by which THBS1 regulated glycolysis and Th17/Treg balance. A total of 12 peripheral blood (4 GO samples, 4 GH samples, and 4 health samples) were collected to measure the percentage of Th17/Treg in monocytes by flow cytometry. RNA sequencing (RNA-seq) combined with MeRIP sequencing (MeRIP-seq) was used to screen differentially expressed and methylated genes. MeRIP-qPCR was performed to evaluate the m6A abundance of THBS1 after WTAP silencing. Glycolysis of CD4+ T cells was reflected by the lactate content and glucose uptake. The number of Th17 cells was increased in GO peripheral blood, whereas the Treg cells decreased. RNA-seq acquired 679 differentially expressed genes (308 up-regulated, and 371 down-regulated) in the CD4+ T cells of GO compared to healthy control. MeRIP-seq identified 3277 m6A peaks between the GO group and the healthy control group, corresponding with 2744 genes (1143 hypermethylated and 1601 hypomethylated). Combined analysis of RNA-seq and MeRIP-seq showed 81 hypermethylated and up-regulated genes. Among the six candidate genes in the PI3K-signaling pathway, THBS1 was the most significantly differentially expressed and hypermethylated. THBS1 silencing resulted in decreased lactate content and glucose uptake in CD4+ T cells. WTAP was significantly upregulated in CD4+ T cells of GO, and WTAP silencing significantly reduced m6A abundance and expression of THBS1. Upregulated and hypermethylated THBS1 mediated by WTAP promoted glycolysis of CD4+ T cells, affected Th17/Treg balance, and facilitated GO progression. We provided a novel potential target for GO treatment and revealed the molecular mechanism of WTAP and THBS1 in GO under the m6A perspective.https://www.tandfonline.com/doi/10.1080/08916934.2024.2433628Graves’ ophthalmopathyWTAPTHBS1N6-methylationglycolysisPI3K-Akt signaling pathway
spellingShingle Lin-na Li
Jie-man Wu
Zong-ji Zheng
Shu-xian Li
Meng-yi Cai
Meng-chen Zou
N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
Autoimmunity
Graves’ ophthalmopathy
WTAP
THBS1
N6-methylation
glycolysis
PI3K-Akt signaling pathway
title N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
title_full N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
title_fullStr N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
title_full_unstemmed N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
title_short N6-methyladenosine modification of THBS1 induced by affluent WTAP promotes Graves’ ophthalmopathy progression through glycolysis to affect Th17/Treg balance
title_sort n6 methyladenosine modification of thbs1 induced by affluent wtap promotes graves ophthalmopathy progression through glycolysis to affect th17 treg balance
topic Graves’ ophthalmopathy
WTAP
THBS1
N6-methylation
glycolysis
PI3K-Akt signaling pathway
url https://www.tandfonline.com/doi/10.1080/08916934.2024.2433628
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