Potential effects of cuproptosis-related genes in the onset of ankylosing spondylitis: a multi-omics Mendelian randomization study

doi:10.12307/2026.734

Abstract

Abstract: BACKGROUND: Cuproptosis may be involved in the pathogenesis of ankylosing spondylitis, but there is a lack of direct genetic evidence.
OBJECTIVE: To investigate the genetic relationship between cuproptosis-related genes and ankylosing spondylitis using multi-omics Mendelian randomization analysis, heterogeneity testing, and colocalization methods.
METHODS: The genome-wide association study (GWAS) data related to ankylosing spondylitis were obtained from the FinnGen database (jointly developed by Finland and the University of Helsinki), the UK Biobank (UKB) database (jointly established by the UK Medical Research Council and the UK government), and the GWAS Catalog database (developed by the European Molecular Biology Laboratory and the European Bioinformatics Institute). All these databases are open access, and the studies involved have been approved by the relevant institutional review boards. We acquired quantitative trait loci data related to ankylosing spondylitis, specifically focusing on blood methylation, gene expression, and protein abundance. We used data from FinnGen as the primary discovery set, further validating our findings with supplementary data from the UK Biobank and the GWAS Catalog. The multi-omics Mendelian randomization method helped us assess the potential molecular links between the molecular features of cuproptosis-related genes and ankylosing spondylitis. Finally, we employed colocalization analysis to determine if the detected genetic signals shared common causal variants.
RESULTS AND CONCLUSION: Our analysis revealed potential genetic causal links between ankylosing spondylitis and several genes and loci, including carcinoembryonic antigen-related cell adhesion molecule 8 (cg09422614), ferric chelate reductase 1 (cg09370016), galactose-3-O sulfotransferase 1 (cg04030848), sideroflexin 5 (cg03344820), sulfite oxidase (cg06495347, cg22580629), thymidine phosphorylase (cg11654620, cg16367976). These genes and loci showed significant associations in multi-omics Mendelian randomization analyses involving both blood mQTLs and eQTLs. Colocalization analysis further strengthened the evidence for an association between the methylation loci of galactose-3-O sulfotransferase 1, sulfite oxidase, and thymidine phosphorylase and the risk of ankylosing spondylitis (PP.H4 > 0.5). Our investigation into methylation regulatory mechanisms indicated that increased methylation levels at cg09422614, cg09370016, and cg03344820 were positively correlated with the risk of ankylosing spondylitis. The risk of ankylosing spondylitis was increased by positively regulating the expression levels of these genes. Conversely, while methylation levels at cg04030848, cg06495347, and cg22580629 also showed a positive correlation with the risk of ankylosing spondylitis, and they appeared to increase the risk of ankylosing spondylitis by positively regulating the expression levels of their respective genes. Interestingly, thymidine phosphorylase gene expression levels showed a negative correlation with the risk of ankylosing spondylitis, with methylation at cg11654620 and cg16367976 positively regulating thymidine phosphorylase gene expression. The GSE25101 dataset blood sample analysis indicated that carcinoembryonic antigen-related cell adhesion molecule 8, ferric chelate reductase 1, galactose-3-O sulfotransferase 1, sideroflexin 5, sulfite oxidase, and thymidine phosphorylase exert regulatory effects in the pathogenesis of ankylosing spondylitis and may participate in its pathological progression. This study not only sheds light on the potential pathogenic role of cuproptosis mechanisms in ankylosing spondylitis but also offers a theoretical basis for future multi-omics functional mechanism research and precision diagnosis and individualized treatment of ankylosing spondylitis, particularly within the Chinese population.

Key words: ankylosing spondylitis, Mendelian randomization, cuproptosis, colocalization, causal association, single nucleotide polymorphism

CLC Number:

Song Zhichao, Qi Wenrong, Meng Peng, Zhang Yanyan. Potential effects of cuproptosis-related genes in the onset of ankylosing spondylitis: a multi-omics Mendelian randomization study[J]. Chinese Journal of Tissue Engineering Research, 2026, 30(16): 4240-4252.

Figures/Tables 14

2.3 整合强直性脊柱炎全基因组关联分析和铜死亡相关的血液pQTL数据铜死亡相关蛋白丰度与强直性脊柱炎的因果关系见表5，共鉴定出7个铜死亡相关蛋白与强直性脊柱炎风险相关(PSMR < 0.05，PSMR multi < 0.05，PHEIDI > 0.05)。这7个蛋白在染色体上的分布情况见图6。除丙氨酸乙醛酸转氨酶(AGXT)、硫酸酯酶修饰因子1 (SUMF1)、核因子kB亚基1(NFKB1)外，其余蛋白丰度与强直性脊柱炎风险均呈负相关，见图7。共定位分析结果显示，上述鉴定到的特征信号中核因子kB亚基1和胸苷磷酸化酶(TYMP)在共定位区域窗口中具有较强的共定位证据支持(PP.H4 > 0.5)，共定位结果图见图8。验证结果显示，核因子kB亚基1(PSMR=6.67×10-3，PSMR multi=3.13×10-2，PHEIDI=0.63)、一氧化氮合酶2(NOS2)(PSMR=8.75×10-3，PSMR multi=3.24×10-2，PHEIDI=0.25)共2个编码蛋白在UKB队列中得到验证，但没有蛋白在GCST90044542队列中得到验证。 2.4 整合强直性脊柱炎全基因组关联分析、铜死亡相关的血液mQTL和eQTL数据基于铜死亡相关血液eQTL、mQTL与强直性脊柱炎全基因组关联分析的多组学数据孟德尔随机化分析中所得关键结果，作者认为铁螯合还原酶1、侧链蛋白5、SLC25A13、SLC29A2、SLC6A12、亚硫酸氧化酶、花生四烯酸-12-脂加氧酶、信号转导和转录激活因子(STAT5B)、SLC16A5、癌胚抗原相关细胞附着分子8、半乳糖-3-O-磺基转移酶1、胸苷磷酸化酶基因可能与强直性脊柱炎有因果关联。进一步以mQTL作为暴露条件，eQTL作为结局的情况下进行多组学数据孟德尔随机化分析，以探索上述交叉结果中DNA甲基化位点甲基化对所在基因表达是否具有显著的调控作用。结果发现癌胚抗原相关细胞附着分子8(cg09422614)、铁螯合还原酶1(cg09370016)、半乳糖-3-O-磺基转移酶1(cg04030848)、侧链蛋白5(cg03344820)、亚硫酸氧化酶(cg06495347、cg22580629)、胸苷磷酸化酶(cg11654620、cg16367976)在"

上述结果中均得到了显著验证(PSMR < 0.05，PSMR multi < 0.05，PHEIDI > 0.05)，见表6。 2.5 整合强直性脊柱炎全基因组关联分析、铜死亡相关的血液eQTL和pQTL数据综合考虑铜死亡相关血液pQTL与强直性脊柱炎全基因组关联分析的多组学数据孟德尔随机化分析结果，作者认为胸苷磷酸化酶基因可能与强直性脊柱炎有因果关联。进一步的在血液eQTL为暴露条件，pQTL为结局的情况下进行多组学数据孟德尔随机化分析，以探索上述交叉结果中基因对所在蛋白丰度是否具有显著的调控作用。结果显示多组学数据孟德尔随机化基因未得到显著验证。 2.6 整合多组学水平证据基于以上分析结果，作者认为癌胚抗原相关细胞附着分子8(cg09422614)、铁螯合还原酶1(cg09370016)、半乳糖-3-O-磺基转移酶1(GAL3ST1，cg04030848)、侧链蛋白5(cg03344820)、亚硫酸氧化酶(cg06495347、cg22580629)、胸苷磷酸化酶(cg11654620、cg16367976)可能与强直性脊柱炎有因果关联，所述关键结果位点、基因在染色体上的分布情况见图9。上述基因在血液mQTL、eQTL的多组学数据孟德尔随机化分析中均验证显著。半乳糖-3-O-磺基转移酶1的甲基化位点cg04030848在mQTL与强直性脊柱炎全基因组关联分析的共定位分析中得到了较强共定位证据支持(PP.H4 > 0.5)，亚硫酸氧化酶和胸苷磷酸化酶则在eQTL与强直性脊柱炎全基因组关联分析的共定位分析中得到了较强共定位证据支持(PP.H4 > 0.5)。上述基因及位点在mQTL-eQTL分析中均得到显著性验证。通过OR值对风险相关性及调控方向进行判断，结果发现cg09422614、cg09370016和cg03344820位点的甲基化水平与强直性脊柱炎风险呈正相关，癌胚抗原相关细胞附着分子8、铁螯合还原酶1和侧链蛋白5基因的表达水平与强直性脊柱炎风险呈负相关，因此，cg09422614、cg09370016和cg03344820位点甲基化分别负向调控各自对应的基因表达水平。而cg04030848、cg06495347和cg22580629位点的甲基化水平与强直性脊柱炎风险呈正相关，半乳糖-3-O-磺基转移酶1和亚硫酸氧化酶基因的表达水平与强直性脊柱炎风险呈正相关，因此，cg04030848、cg06495347和cg22580629位点甲基化分别正向调控所在基因的表达水平。此外，还发现cg11654620和cg16367976位点的甲基化水平与强直性脊柱炎风险呈负相关，而胸苷磷酸化酶基因的表达水平则与强直性脊柱炎风险呈负相关，因此，cg11654620和cg16367976位点甲基化正向调控胸苷磷酸化酶基因的表达水平。综上，可能存在假定的机制：cg09422614、cg09370016和cg03344820位点较高的甲基化水平分别抑制了癌胚抗原相关细胞附着分子8、铁螯合还原酶1和侧链蛋白5基"

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