Jaswani Pradeep, Prakash Swayam, Agrawal Suraksha, Prasad Narayan, Sharma Raj Kumar
Department of Nephrology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Raibareily Road, Lucknow- 226014, U.P, India.
Department of Hematology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Raibareily Road, Lucknow-226014, U.P, India.
Microrna. 2017 Dec 6;6(3):213-221. doi: 10.2174/2211536606666171016151846.
Translational research on miRNAs develops reliable biomarkers for diagnosis and prognosis of renal diseases. Bioinformatic analyses and systems biology could drive the research for knowing new informative miRNA targets.
This study proposes an approach to identify miRNA specific significant target genes, and single nucleotide polymorphisms (SNPs) associated with renal pathophysiology.
miRNAs were selected after removing duplicity, on the basis of techniques used, and disease spectrum width score. Target genes were predicted from different databases like miRWalk, miRTarBase, and DIANA-TarBase. SNPs were prioritized on the basis of target score and conserved energy score available in MirSNP database. miRNAs were characterized as "specific", "strong", "likely", "unlikely", and "irrelevant" biomarkers. PCR-SSP based genotyping was carried out to access the molecular profiling of hsa-miR-192 and TGF-β1 followed by quantitative real time PCR to analyze expression level of TGF-β1. The relative expression levels of mRNAs were analyzed by 2-ΔΔCt method.
170 renal associated miRNAs were found to be up-regulated, down-regulated or differentially expressed. Noticeably hsa-miR-192-3p expression was reported in nine diseases. 117 genes were associated with basic kidney diseases and end stage renal disease (ESRD). Threshold > 80% for 93 target genes was observed from mirSVR. Mutant genotypes for hsa-miR-192 (OR=4.64, p-value ≤ 0.0001) and its corresponding target gene TGF-β1 (OR = 0.70, p-value = 0.0351) showed susceptible association with ESRD. More so, patients possessing mutant allele of TGF-β1 showed elevated mRNA expression (Fold change = 9.83).
Study proposed a new approach to identify specific miRNA biomarkers for particular diseases with corresponding target genes and SNPs and also highlighted the importance of hsa-miR-192 in renal diseases.
微小RNA（miRNA）的转化研究为肾脏疾病的诊断和预后开发可靠的生物标志物。生物信息学分析和系统生物学可推动了解新的信息丰富的miRNA靶标的研究。
本研究提出一种方法来鉴定miRNA特异性的重要靶基因以及与肾脏病理生理学相关的单核苷酸多态性（SNP）。
根据所使用的技术和疾病谱宽度评分，去除重复后选择miRNA。从不同数据库如miRWalk、miRTarBase和DIANA-TarBase预测靶基因。根据MirSNP数据库中可用的靶标分数和保守能量分数对SNP进行优先级排序。将miRNA表征为“特异性”、“强”、“可能”、“不太可能”和“不相关”生物标志物。进行基于PCR-SSP的基因分型以获取hsa-miR-192和转化生长因子-β1（TGF-β1）的分子谱，随后进行定量实时PCR以分析TGF-β1的表达水平。通过2-ΔΔCt法分析mRNA的相对表达水平。
发现170个与肾脏相关的miRNA上调、下调或差异表达。值得注意的是，hsa-miR-192-3p在9种疾病中被报道有表达。117个基因与基础肾脏疾病和终末期肾病（ESRD）相关。从mirSVR观察到93个靶基因的阈值>80%。hsa-miR-192的突变基因型（OR=4.64，p值≤0.0001）及其相应的靶基因TGF-β1（OR = 0.70，p值 = 0.0351）显示与ESRD有易感性关联。更重要的是，携带TGF-β1突变等位基因的患者显示mRNA表达升高（倍数变化 = 9.83）。
本研究提出了一种新方法来鉴定特定疾病的特异性miRNA生物标志物及其相应的靶基因和SNP，同时强调了hsa-miR-192在肾脏疾病中的重要性。
