Cell-free MicroRNA miR-505-3p in Graft Preservation Fluid Is an Independent Predictor of Delayed Graft Function After Kidney Transplantation

Abstract

Background Delayed graft function (DGF), a common complication after transplantation of deceased donor kidneys, affects both short- and long-term outcomes. Currently available biomarkers during graft preservation lack sensitivity in predicting risk for DGF. The aim of this study is to identify cell-free micro ribonucleic acid (miRNA) biomarkers in graft preservation fluid predictive of DGF after kidney transplantation. Methods Vascular bed preservation fluid was collected from 48 kidney grafts from donation after circulatory death (DCD) or donation after brain death (DBD) donors. miRNA profiles were determined by polymerase chain reaction (PCR) array (n = 8) and validated by reverse transcription and quantitative PCR (n = 40). Graft function posttransplantation was defined as immediate good function (IF) or DGF. Results A total of 223 miRNAs fulfilled the preset parameters (Ct < 40 in 3 or more samples) and were included in the analysis. Thirty-two miRNAs were significantly different between DGF and IF kidney grafts (P < 0.05) but, after correction for multiple testing, only miR-505-3p remained significant. The significant association of high miR-505-3p levels with DGF was confirmed in an independent validation cohort using conventional reverse transcription and quantitative PCR detection. Multivariate analyses showed miR-505-3p as an independent predictor for DGF (odds ratio, 1.12; P = 0.028). If stratified for donor type, miR-505-3p levels remained significantly different between IF and DGF in DCD grafts (P < 0.01), but not in DBD grafts. Receiver operating characteristic curve analysis showed a high sensitivity and specificity (area under the curve, 0.833). Conclusions In DCD grafts, high levels of miR-505-3p in preservation fluid are associated with increased risk of DGF after kidney transplantation. Further study is required to confirm the utility of cell-free miR-505-3p as prognostic biomarker for DGF.

Identification and Validation Model for Informative Liquid Biopsy-Based microRNA Biomarkers: Insights from Germ Cell Tumor In Vitro, In Vivo and Patient-Derived Data

“…Importantly, we believe the major reason for the little impact of hemolysis on specific targets in our study is the distinct microRNA isolation procedure: we have performed a magnetic bead capture of microRNAs, which are isolated and purified from the serum contents, eliminating the potentially harmful effects of molecules such as hemoglobin or heparin, which are known to inhibit the PCR reaction [60][61][62][63]. Indeed, we have witnessed this effect in RNA isolated from kidney preservation fluids (described in [54]) containing heparin contamination, a commonly used anticoagulant. Our new results again demonstrate that in the bead-based capture no inhibitory effect of heparin was observed and heparinase 1 digestion did not reduce the Ct values of both endogenous and spiked-in microRNAs (Supplementary Figure S12).…”

“…For exploring the impact of heparin contamination in microRNA quantification, a subset of graft preservation fluids (n = 8) from patients undergoing kidney transplantation (described by us in [54]) that were shown to be contaminated with heparin (n = 4) was selected, and subsequently quantified after microRNA isolation using miRNEAsy spin columns or after bead capture followed by pre-amplification. Ct values were determined and compared with or without heparinase 1 treatment.…”

Recent Advances on Biomarkers of Early and Late Kidney Graft Dysfunction

“…They can be isolated, quantified and profiled by multiple platforms which can also characterize their target genes [ 32 ]. They have been studied in graft preservation fluid and proposed as viability biomarkers (miR-486-5p, miR-144-3p, miR-142-5p, and miR-144-5p) [ 12 ]; however, only miR-505-3p has been demonstrated to be an independent predictor of DGF in DCD grafts with high accuracy (AUC = 0.83) and was confirmed in a validation cohort [ 13 ].…”

Diagnostic, Prognostic, and Therapeutic Value of Non-Coding RNA Expression Profiles in Renal Transplantation

“…Interestingly, cell-free miRNAs in graft preservation fluid are predictive of DGF post-transplantation. Roest HP et al identified an association between high levels of miR-505-3p in the preservation fluid of kidney grafts donated after circulatory death and an increased risk of DGF after transplantation [43]. Another prospective cohort study that analyzed graft dysfunction in transplant recipients of expanded criteria donor organs confirmed the significance of a subset of four miRNAs (miR-486-5p, miR-144-3p, miR-142-5p, and miR-144-5p) previously identified in DGF development [44].…”