MicroRNAs as a therapeutic target in IgA nephropathy in Indian population
Immunoglobulin A Nephropathy (IgAN) is the most occurring glomerular disease with rapid development to End Stage Renal Disease requiring renal replacement therapy. Genome-wide studies suggest geographical variations in genetic susceptibility to IgAN and disease progression have been shown to play a vital role in physio pathological state. In this study, we have identified new microRNAs allied with IgAN in Indian population. The miRNA was isolated from kidney biopsies of IgAN patients and healthy control (healthy tissue from patients with renal cell carcinoma). Small RNA library preparation was performed, followed by Next Generation Sequencing on the ION S5 Torrent at 1x50 base pairs. Later the miRNA expression analyses were carried and obtained high-quality reads from BAM file. The sequence of miRNA reads was mapped to reference Human genome (hg38). The Partek Flow Genomic Analysis Software was used to compare the miRNAs expression between healthy participants and patients with IgAN.
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It is a single centre hospital-based case control, retrospective study which was conducted at AIG (Mayo Clinic Care Network) hospital, Hyderabad, India from February 2021 to April 2022 that includes the kidney biopsy blocks of patients with IgAN (n=6) and controls (n=6) from patients with renal cell carcinoma (healthy tissue). The clinical features of IgAN patients after diagnosed by experienced pathologists, are shown in the manuscript. Following steps were involved in the process of samples and data analysis 1. FFPE blocks of the biopsy specimens were collected and miRNA was isolated. 2. Library preparation for small RNA sequencing: CleanTag™ Small RNA Library Preparation Kit was used. Purified library products were evaluated using the High Sensitivity DNA Chip for Agilent 2100 Tape Station (Agilent Technologies, Inc.), followed by Next Generation Sequencing on the ION S5 Torrent at 1x50 base pairs. 3. Quality control and reads mapping to the reference genome: BAM file of raw reads were processed and high-quality reads were obtained. The sequence of miRNA reads was mapped to reference Human genome (hg38) by Bowtie2 (v2.2) using Partek Flow Genomic Analysis Software.