MicroRNA Expression Profiling In Serum Of Acute Renal Allograft Rjection

Background:Renal transplantation is the most ideal and oecumenical treatment of choice for end-stage renal disease (ESRD) patients.But transplantation rejection is still a strong risk factor for recipients of renal grafts. Acute rejection (AR),especially,is a significant cause of allograft loss and frequently contributes to chronic allograft dysfunction.Rejection may be either cellular or antibody mediated.The detection of acute rejection,regardless of etiology,is critically dependent on measurement of serum creatinine,an insensitive measure of renal injury.Ultimately,a kidney biopsy is the gold standard but is invasive, is limited by sampling error.Thus,sensitive and less invasive methods would be clinically useful for detection of AR.Moreover, while such methods may be diagnostic,it would also be beneficial if they can also be used tomonitor immunologic activity prior to established disease.MicroRNAs (miRNAs) are short (usually 18-22 nucleotides),endogenous non-coding RNAs (ncRNAs) that inhibit gene expression post-transcriptionally in a sequence-specific mode.miRNAs are epigenetic regulators of gene expression that are able to modulate several cellular processes, from development to disease conditions. Our study objective was to compare the levels of miRNAs expression in serum of the group with AR between the group without AR.In order to find a new biological markers for clinical diagnosis AR.Objectives:The microarray analysis was used to identify the microRNAs profile related with acute rejection(AR) after renal transplantation,The microRNAs screened by microarray analysis were validated by RT-PCR using the serum sample.Methods:l.Screening microRNA makers correlated with Acute Renal Allograft RjectionThe total RNA from serum of 3 pairs of renal transplantation patients with or without acute rejection.The different profiles of microRNA between two groups were acquired by the Agilent microarray to identify the microRNA makers correlated with acute renal allograft rejection.2.Clinical validation of screened microRNAs correlated with Acute Renal Allograft RjectionCombining the result of microarray analysis and conclusions of published studies,three microRNA(miR-142-5p, miR-144-5p, miR-130a-3p) were validated by quantity RT-PCR using serum samples of 30 patients:10 of them with AR,10 of them without AR and the rest were nomal control group.The impact of above listed microRNAs on the occurrence of acute renal allograft rejection were analyzed.Results:The analysis of microarray showed that 20 microRNAs had significant different expression between with and without AR groups.Furthermore,the upregulation of miR-142-5p, miR-144-5p and miR-130a-3p in AR group were validated by RT-PCR using serum sample.Conclusions:1.The use of microarray technology to screen, combined with qPCR technology to verify the experimental results, is the effective approach for the analysis of miRNAs expression profile in acute rejection of renal transplantation2.3 miRNAs were selected for relative quantification by RT-PCR.All results was conformable to the result of array which confirmed the reliability of Microarray chip results.miR-142-5p, miR-144-5p and miR-130a-3p are correlated with acute renal allograft rejection.3.There were 20 miRNAs significant differential expression in acute rejection after renal transplantation by Microarray chip,13 of them were upregulated and 7 of them were downregulated.