School of Public Health and Graduate institute of Life Sciences, National Defense Medical Center, No.161, Sec. 6, Minquan E. Rd., Neihu Dist., Taipei City, 114, Taiwan, Republic of China.
Division of Nephrology, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, Taipei City, Taiwan, Republic of China.
BMC Nephrol. 2019 Aug 5;20(1):300. doi: 10.1186/s12882-019-1471-2.
A chronic inflammatory state is a prominent feature in patients with end-stage renal disease (ESRD). Nuclear factor-kappa B (NF-κB) is a transcription factor that regulates the expression of genes involved in inflammation. Some genetic studies have demonstrated that the NF-κB genetic mutation could cause kidney injury and kidney disease progression. However, the association of a gene polymorphism in the transcription factor binding site of NF-κB with kidney disease is not clear.
We used the Taiwan Biobank database, the University of California, Santa Cruz, reference genome, and a chromatin immunoprecipitation sequencing database to find single nucleotide polymorphisms (SNPs) at potential binding sites of NF-κB. In addition, we performed a case-control study and genotyped 847 patients with ESRD and 846 healthy controls at Tri-Service General Hospital from 2015 to 2016. Furthermore, we used the ChIP assay to identify the binding activity of different genotypes and used Luciferase reporter assay to examine the function of the rs9395890 polymorphism.
The results of biometric screening in the databases revealed 15 SNPs with the potential binding site of NF-κB. Genotype distributions of rs9395890 were significantly different in ESRD cases and healthy controls (P = 0.049). The ChIP assay revealed an approximately 1.49-fold enrichment of NF-κB of the variant type TT when compared to that of the wild-type GG in rs9395890 (P = 0.027; TT = 3.20 ± 0.16, GT = 2.81 ± 0.20, GG = 1.71 ± 0.18). The luciferase reporter assay showed that the NF-κB binding site activity in T allele was slightly higher than that in G allele, though it is not significant.
Our findings indicate that rs9395890 is associated with susceptibility to ESRD in Taiwan population.
慢性炎症状态是终末期肾病（ESRD）患者的一个显著特征。核因子-κB（NF-κB）是一种转录因子，可调节参与炎症的基因表达。一些遗传研究表明，NF-κB 基因突变可导致肾脏损伤和肾脏疾病进展。然而，NF-κB 转录因子结合位点的基因多态性与肾脏疾病的关系尚不清楚。
我们使用台湾生物银行数据库、加利福尼亚大学圣克鲁兹分校参考基因组和染色质免疫沉淀测序数据库，在 NF-κB 的潜在结合位点处寻找单核苷酸多态性（SNP）。此外，我们进行了病例对照研究，在 2015 年至 2016 年期间，对三军总医院的 847 名 ESRD 患者和 846 名健康对照者进行了基因分型。此外，我们使用 ChIP 测定法来鉴定不同基因型的结合活性，并使用荧光素酶报告基因测定法来检测 rs9395890 多态性的功能。
数据库中的生物特征筛选结果显示，有 15 个具有 NF-κB 潜在结合位点的 SNP。rs9395890 的基因型分布在 ESRD 病例和健康对照组之间存在显著差异（P=0.049）。ChIP 测定显示，与 rs9395890 中的野生型 GG 相比，变异型 TT 的 NF-κB 富集约 1.49 倍（P=0.027；TT=3.20±0.16，GT=2.81±0.20，GG=1.71±0.18）。荧光素酶报告基因测定显示，T 等位基因的 NF-κB 结合位点活性略高于 G 等位基因，但无统计学意义。
我们的研究结果表明，rs9395890 与台湾人群 ESRD 的易感性有关。
