Zhang Wanyun, Zhang Pei, Pu Yanlin, Chen Zhijun, Su Guannan, Deng Yang, Zhang Yinan, Ji Yan, Huang Ziqian, Zhou Qian, Luo Xiang, Lai Yujie, Yang Peizeng
Ophthalmology Medical Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing Key Laboratory for the Prevention and Treatment of Major Blinding Eye Diseases, Chongqing Branch (Municipality Division) of National Clinical Research Centre for Ocular Diseases, Chongqing, China.
Department of Ophthalmology, Bishan Hospital of Chongqing, Bishan hospital of Chongqing medical university, Chongqing, China.
Invest Ophthalmol Vis Sci. 2025 Aug 1;66(11):28. doi: 10.1167/iovs.66.11.28.
Behçet's uveitis (BU) is an auto-inflammatory disease frequently with a poor prognosis. Here, we performed an integrated analysis of DNA 5-hydroxymethylcytosine (5-hmC) landscape and transcriptomic profiling in CD4+ T cells from active BU patients and healthy controls.
We conducted an integrated analysis of DNA 5-hmC modifications and transcriptomic data from CD4+ T cells of active BU patients and healthy individuals. Publicly available single-cell RNA-sequencing data were analyzed to validate findings across cell clusters. Functional experiments were performed to assess the effects of TUBB4B on cytokine production and T-cell frequencies.
Bioinformatics analyses identify 801 downregulated and 2489 upregulated differential hydroxymethylated genes, predominantly enriched in pathways related to pathogen defense and various immune responses. RNA sequencing reveals 958 downregulated and 572 upregulated genes, with significant enrichment in pathways related to bacterial infection. Integration of epigenetic and transcriptional data highlights 74 candidate genes and three genes including TUBB4B, SKI and ZFPM1 are validated to be downregulated in active BU patients. Further analysis of publicly available single-cell RNA-sequencing data reveals TUBB4B downregulation across multiple cell clusters, particularly in naïve CD4+ T cells. Functional experiments indicate that overexpressing TUBB4B could decrease the frequencies of IL-17+ and IFN-γ+ CD4+ T cells and reduce the production of IL-17 and IFN-γ, while upregulating the frequency of CD4+CD25+FOXP3+ T cells and enhancing secretion of IL-10.
This study underscores the pivotal role of pathogen-induced immune dysregulation in BU development and identifies TUBB4B as a potential therapeutic target for the study on prevention and treatment of this disease.
白塞氏葡萄膜炎（BU）是一种自身炎症性疾病，预后通常较差。在此，我们对活动期BU患者和健康对照者的CD4+T细胞中的DNA 5-羟甲基胞嘧啶（5-hmC）图谱和转录组分析进行了综合分析。
我们对活动期BU患者和健康个体的CD4+T细胞的DNA 5-hmC修饰和转录组数据进行了综合分析。分析公开可用的单细胞RNA测序数据以验证跨细胞簇的研究结果。进行功能实验以评估TUBB4B对细胞因子产生和T细胞频率的影响。
生物信息学分析鉴定出801个下调和2489个上调的差异羟甲基化基因，主要富集在与病原体防御和各种免疫反应相关的途径中。RNA测序揭示了958个下调和572个上调的基因，在与细菌感染相关的途径中显著富集。表观遗传和转录数据的整合突出了74个候选基因，包括TUBB4B、SKI和ZFPM1在内的三个基因被证实在活动期BU患者中下调。对公开可用的单细胞RNA测序数据的进一步分析揭示了TUBB4B在多个细胞簇中下调，特别是在初始CD4+T细胞中。功能实验表明，过表达TUBB4B可降低IL-17+和IFN-γ+CD4+T细胞的频率，减少IL-17和IFN-γ的产生，同时上调CD4+CD25+FOXP3+T细胞的频率并增强IL-10的分泌。
本研究强调了病原体诱导的免疫失调在BU发病中的关键作用，并确定TUBB4B是该疾病预防和治疗研究的潜在治疗靶点。
