Expression and 7-day time course of circulating microRNAs in septic patients treated with nephrotoxic antibiotic agents

Abstract

Background: Through regulation of signaling pathways, microRNAs (miRNAs) can be involved in sepsis and associated organ dysfunction. The aims of this study were to track the 7-day time course of blood miRNAs in patients with sepsis treated with vancomycin, gentamicin, or a non-nephrotoxic antibiotic and miRNA associations with neutrophil gelatinase-associated lipokalin (NGAL), creatinine, procalcitonin, interleukin-6, and acute kidney injury (AKI) stage.

Methods: Of 46 adult patients, 7 were on vancomycin, 20 on gentamicin, and 19 on another antibiotic. Blood samples were collected on days 1, 4, and 7 of treatment, and miRNAs were identified using quantitative reverse transcription PCR.

Results: The results showed no relationship between miRNA levels and biochemical variables on day 1. By day 7 of gentamicin treatment miR-15a-5p provided good discrimination between AKI and non-AKI (area under curve, 0.828). In patients taking vancomycin, miR-155-5p and miR-192-5p positively correlated with creatinine and NGAL values, and miR-192-5p and miR-423-5p positively correlated with procalcitonin and interleukin-6 in patients treated with a non-nephrotoxic antibiotic. In patients together we found positive correlation between miR-155-5p and miR-423-5p and all biochemical markers.

Conclusion: The results suggest that these four miRNAs may serve as diagnostic or therapeutic tool in sepsis, renal injury and nephrotoxic treatment.

Trial registration: ClinicalTrials.gov , ID: NCT04991376 . Registered on 27 July 2021.

Keywords: Acute kidney injury; Gentamicin; Nephrotoxicity; Sepsis; Vancomycin; microRNA.

Conflict of interest statement

The authors declare that they have no competing interests related to this manuscript or project.

© 2022. The Author(s).

Figures

Fig. 1

Similar pattern of individual expression of four miRNAs in each treatment group on days 1, 4, and 7 of treatment. a Vancomycin group, b gentamicin group, and c other ATB group. The figures show the log 2-transformed label-free quantification of the four investigated miRNAs during the treatment period. P values are from Kruskal–Wallis test, and **** indicates p < 0.001 between the miRNAs themselves. Post-hoc analysis was performed using Dunn’s method with Bonferroni correction. Each investigated miRNA presented a similar respective pattern of expression across all groups

Fig. 2

Time-dependent change in expression of miR-15-5p and miR-423-5p in the other-antibiotic group and the gentamicin group (vancomycin not shown). Log 2–transformed label-free quantification of differential expression of two miRNAs. amiR-15-5p in the other-ATB group. The figure shows a significant change in expression of miR-15a-5p between days 1 and 4 and days 4 and 7. bmiR-423-5p in the gentamicin group, in which a statistically significant change was detected between days 4 and 7. Data are presented as medians and interquartile ranges. P values are the result of the Friedman test and post-hoc Wilcoxon test

Fig. 3

Principal component analysis of the relationship between miRNAs and biochemical variables on study day 1. Principal component analysis biplot shows log 2–transformed label-free quantification of the concentrations of variables and the difference between miRNA expression and renal injury or inflammation markers on day 1 in all patients, regardless of subsequent treatment. No relationship was identified between miRNAs and biochemical variables

Fig. 4

miRNA expression in patients with and without acute kidney injury (AKI) on day 7. Log 2–transformed label-free quantification of differential expression of miRNAs in patients with and without acute kidney injury in all three groups (vancomycin, gentamicin, other ATB) on day 7 is presented on the figure. The statistically significant difference between patients with and without acute kidney injury is observed in miR-15a-5p in gentamicin group. * - p value of Mann–Whitney U test (p = 0.013)

Fig. 5

miR-15a-5p may be a biomarker for acute kidney injury (AKI) in patients with sepsis treated by gentamicin. Data are presented as log 2–transformed label-free quantification of miRNA serum concentrations and expression. a Serum concentrations of miR-15a-5p in patients with AKI versus non-AKI in the gentamicin group; p values are from the Mann–Whitney U test. b Relative receiver operating characteristic (ROC) curve with the area under the curve obtained from by simple logistic regression; ** log-likelihood ratio test, p = 0.008

Fig. 6

Simplified graphical presented associations between sepsis, nephrotoxic antibiotics, investigated miRNAs and their target regulatory genes in sepsis induced acute kidney injury. On the figure are presented simplified associations between gentamicin, vancomycin, endotoxin, ischemia/reperfusion injury and miR-15a-5p, miR-423-5p, miR-192-5p and miR-155-5p with their previously established target genes. Acute kidney injury in sepsis is a complex process involving combination of risk factors as ischemia, inflammation and possible nephrotoxicity. Abbreviations: AKI – acute kidney injury, Bax – B-cell lymphoma 2 Asociated X, Apoptosis Regulator, DDIT4 – DNA damage–inducible transcript 4, FOXA1 – forkhead box protein A1, GSTM1 – glutathione-S-transferase M1, IFR2BP2-NFAT1 – interferon regulatory factor 2 binding protein 2–nuclear factor of activated T cells 1, MAFF – MAF basic leucine zipper transcription factor F, miR – micro RNA, MYLK – myosin light chain kinase, PRG2 – proteoglycan 2, pro eosinophil, major basic protein, p21– p21 gene, p53 – p53 gene, TNIP2 –TNFAIP3 interacting protein 2 gene, VEGFA – vascular endothelial growth factor A gene, VEGFC – vascular endothelial growth factor C gene, XIAP – X-linked inhibitor of apoptosis, XIST-CUL3 – X inactive specific transcript - cullin 3 gene