Department of Medical Biology, Faculty of Medicine, Hacettepe University, Ankara, Turkey.
Division of Rheumatology, Department of Pediatrics, Faculty of Medicine, Hacettepe University, Ankara, Turkey.
Int Immunopharmacol. 2023 Nov;124(Pt B):111011. doi: 10.1016/j.intimp.2023.111011. Epub 2023 Oct 14.
Colchicine is the primary treatment for familial Mediterranean fever (FMF). Although colchicine is safe and effective in FMF patients, around 5-10% of patients show resistance to the drug. This study investigates the possibility of a link between colchicine resistance and the distinct miRNA profiles in colchicine resistant FMF patients.
Differentially expressed miRNAs in colchicine resistant FMF patients were detected by Affymetrix 4.0 miRNA array analysis. These miRNAs were then categorized based on the role of their target genes in drug metabolism and inflammation related pathways. qRT-PCR was used to validate candidate miRNAs selected by Enrichr, a gene enrichment analysis system based on the relevance of possible target genes in drug metabolism pathways. Expression levels of these miRNAs' potential target genes were investigated by qRT-PCR. Then, a colchicine resistant hepatoblastoma cell line (HEPG2) was established, and the differentially expressed miRNAs and genes identified in patients were also analyzed in this colchicine-resistant cell line.
25 differentially expressed miRNAs were detected in colchicine resistant FMF patients. miR-183-5p, miR-15b-5p, miR-505-5p, and miR-125a-5p were identified to be associated with drug resistance and inflammatory pathways and thus chosen for further validation. miR-183-5p, miR-15b-5p, miR-505-5p miRNAs showed significantly differential expression in qRT-PCR. NFKB1, NR3C1, PPARα - drug absorption, distribution, metabolism, and excretion (ADME) genes were predicted to be targeted by these miRNAs. Among these targets, NFKB1 and NR3C1 were differentially over expressed in colchicine resistant FMF patients. These findings were validated in the colchicine resistant hepatoblastoma cell line (HEPG2).
This is the first study evaluating the role of miRNAs in colchicine resistant patients with FMF. Their differential expression may result in resistance to standard colchicine treatment by affecting the expression of genes that take place in drug absorption, distribution, metabolism, and excretion (ADME) or nuclear receptors that regulate ADME genes, thus potentially playing a role in both drug metabolism and inflammation.
秋水仙碱是家族性地中海热（FMF）的主要治疗药物。尽管秋水仙碱在 FMF 患者中安全有效，但约有 5-10%的患者对该药物产生耐药性。本研究探讨了秋水仙碱耐药与秋水仙碱耐药 FMF 患者不同 miRNA 谱之间可能存在联系的可能性。
通过 Affymetrix 4.0 miRNA 阵列分析检测秋水仙碱耐药 FMF 患者中差异表达的 miRNA。然后根据其靶基因在药物代谢和炎症相关途径中的作用对这些 miRNA 进行分类。使用基于可能靶基因在药物代谢途径中的相关性的基因富集分析系统 Enrichr 对候选 miRNA 进行 qRT-PCR 验证。通过 qRT-PCR 研究这些 miRNA 潜在靶基因的表达水平。然后，建立了秋水仙碱耐药肝癌细胞系（HEPG2），并对患者中鉴定出的差异表达 miRNA 和基因进行了分析。
在秋水仙碱耐药 FMF 患者中检测到 25 个差异表达的 miRNA。miR-183-5p、miR-15b-5p、miR-505-5p 和 miR-125a-5p 与药物耐药和炎症途径相关，因此被选为进一步验证的对象。miR-183-5p、miR-15b-5p、miR-505-5p 在 qRT-PCR 中显示出明显的差异表达。NFKB1、NR3C1、PPARα-药物吸收、分布、代谢和排泄（ADME）基因被预测为这些 miRNA 的靶标。在这些靶标中，NFKB1 和 NR3C1 在秋水仙碱耐药 FMF 患者中差异过表达。这些发现在秋水仙碱耐药肝癌细胞系（HEPG2）中得到了验证。
这是第一项评估 miRNA 在 FMF 中对秋水仙碱耐药患者作用的研究。它们的差异表达可能通过影响药物吸收、分布、代谢和排泄（ADME）或调节 ADME 基因的核受体的基因表达，从而在药物代谢和炎症中发挥作用，导致对标准秋水仙碱治疗的耐药。
