The calcineurin regulatory subunit polymorphism and the treatment efficacy of tacrolimus for idiopathic membranous nephropathy
The calcineurin regulatory subunit polymorphism and the treatment efficacy of tacrolimus for idiopathic membranous nephropathy
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钙调磷酸酶调节亚基多态性及他克莫司治疗特发性膜性肾病的疗效
DOI：
10.1016/j.intimp.2018.10.038
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发表时间：
2018-12-01
影响因子：
5.6
通讯作者：
Lin, Shanyan
中科院分区：
文献类型：
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作者：
Zhu, Ying;Zhang, Min;Lin, Shanyan
他克莫司被认为是特发性膜性肾病(IMN)的主要治疗选择之一。本研究旨在探讨他克莫司结合蛋白和药物靶点(钙调神经磷酸酶)基因变异与IMN患者疗效的关系及其可能机制。对67例接受他克莫司治疗的IMN患者进行了回顾性研究。对8例IMN患者进行Sanger测序，寻找所有基因外显子的变异，并对59例IMN患者检测到的变异进行基因分型。在人类外周血单核细胞(PBMCs)和其他细胞系中，探索了变异与疗效之间关系的分子机制。编码钙调神经磷酸酶亚单位的PPP3R1基因3‘非翻译区的单核苷酸多态rs875(T>；C)与他克莫司治疗IMN的疗效相关。携带TT基因的患者缓解率(83%)明显高于携带TC/CC基因患者的缓解率(47%，P=0.008)。Western印迹显示TT型携带者外周血中PPP3R1蛋白水平降低(P=0.02)。荧光素酶报告基因分析显示，与C等位基因相比，T等位基因与miR582-5p的结合亲和力增强(P<；0.001)。此外，PPP3R1在Jurkat T细胞系中的敲除增强了他克莫司的免疫抑制作用。我们的研究揭示了PPP3R13‘非编码区rs875基因多态性与IMN患者他克莫司疗效的相关性。该功能多态性可能通过调节miR-582-5p与PPP3R1的相互作用而改变PPP3R1的表达，进而影响他克莫司的免疫抑制作用。
Tacrolimus is considered to be one of the main therapeutic options for idiopathic membranous nephropathy (IMN). This study aimed to investigate the association of variants in genes encoding the binding protein and the drug target (calcineurin) of tacrolimus with the efficacy in IMN patients and the potential mechanism. Sixty-seven IMN patients treated with tacrolimus were enrolled retrospectively. Sanger sequencing was performed to search for variants in all exons of the genes in 8 IMN patients and genotype for the detected variants in the other 59 patients. The molecular mechanism underlying the relationship between the variants and the efficacy was explored in human peripheral blood mononuclear cells (PBMCs) and other cell lines. Single nucleotide polymorphism rs875 (T > C) in the 3'untranslated region (3'UTR) of PPP3R1 encoding calcineurin regulatory subunit was found to be associated with the treatment efficacy of tacrolimus for IMN. Patients carrying TT genotype had a significantly higher remission rate than those carrying TC/CC genotype (83% vs. 47%, P = 0.008). Western blot showed that the TT genotype carriers exhibited reduced PPP3R1 protein levels in PBMCs (P = 0.02). Compared with C allele, T allele displayed increased binding affinity for miR-582-5p in the luciferase reporter assay (P < 0.001). Moreover, knockdown of PPP3R1 in Jurkat T cell line enhanced the immunosuppressive effect of tacrolimus. Our study revealed the association of PPP3R1 3'UTR polymorphism rs875 with the efficacy of tacrolimus in IMN patients. The functional polymorphism might alter PPP3R1 expression via modulating the interaction of miR-582-5p with PPP3R1, which further affected the immunosuppressive effect of tacrolimus.