Related Experiment Videos
Predicting miRNA Association with Corresponding Target Genes and Single Nucleotide Polymorphisms in Altered Renal
Pradeep Jaswani1, Swayam Prakash1, Suraksha Agrawal2
1Department of Nephrology, Sanjay Gandhi Postgraduate Institute of Medical Sciences, Raibareily Road, Lucknow- 226014, U.P, India.
Background:
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.
Objectives:
This study proposes an approach to identify miRNA specific significant target genes, and single nucleotide polymorphisms (SNPs) associated with renal pathophysiology.
Methods:
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.
Results:
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).
Conclusion:
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.