Assessment of a Urinary Kidney MicroRNA Panel as Potential Nephron Segment-Specific Biomarkers of Subacute Renal Toxicity in Preclinical Rat Models

Abstract

Drug-induced kidney injury (DIKI) remains a significant concern during drug development. Whereas FDA-endorsed urinary protein biomarkers encounter limitations including the lack of translatability, there is a considerable interest surrounding the application of microRNAs (miRNAs) in the renal biomarker space. Current knowledge about the value of these novel biomarkers for subacute preclinical rodent studies is still sparse. In this work, Wistar rats were treated with three nephrotoxic compounds-cisplatin (CIS, proximal tubule, 2.5 mg/kg, intraperitoneal [i.p.]), puromycin (PUR, glomerulus, 20/10 mg/kg, i.p.) and N-phenylanthranylic acid (NPAA, collecting ducts, 500 mg/kg, per os)-for up to 28 days to evaluate the performance of a panel of 68 urinary miRNAs as potential nephron segment-specific biomarkers. Out of these 68 kidney injury associated-miRNAs, our selection strategy ultimately revealed rno-miR-34c-5p significantly dysregulated after CIS single administration, and rno-miR-335 and rno-miR-155-5p significantly dysregulated after PUR treatment. In contrast, NPAA daily administration strongly altered the expression profile of 28 miRNAs, with rno-miR-210-3p displaying the most robust changes. A thorough evaluation showed that these miRNA candidates could complement urinary protein biomarkers to detect CIS- or PUR-induced kidney injury in a subacute setting, with a mechanistic (based on rno-miR-34c-5p) and/or a kidney injury detection potential. Our results also provide the first evidence that urinary miRNAs could enhance the detection of collecting duct damage. Overall, these data improve our understanding of the utility of urinary miRNAs as DIKI biomarkers in a subacute DIKI preclinical setting and support the value of using urinary biomarker panels comprising proteins and miRNAs.

Urinary miRNA biomarkers of drug-induced kidney injury and their site specificity within the nephron

“…Additional miRNAs were increased to a greater extent in urine from rats treated with TAL versus proximal tubule toxicants (eg,, which is in agreement with data from LCM experiments showing that these 2 latter miRNAs were overall preferentially enriched in the TAL and/or CD fractions versus the proximal tubules. Interestingly, miR-210-3p was clearly increased in urine from rats treated with TAL toxicants and was also previously reported to be increased in urine from rats treated with the CD toxicant NPAA (Glineur et al, 2018). This is in accordance with results from our localization experiments with LCM samples showing that miR-210-3p was enriched in the TAL and/or CD fractions.…”

“…Six miRNAs were found significantly increased in urine from rats treated with the CD toxicant NPAA in this study. On the other hand, Glineur et al (2018) reported that the expression profiles of 36 different urinary miRNAs were significantly altered in male Wistar rats treated with NPAA at 500 mg/kg/day for up to 28 days, with miR-210-3p displaying the most robust changes. These discrepancies may be explained at least in part by a higher susceptibility of Wistar rats to the kidney lesions induced by NPAA (Betton et al, 2012) as compared with SD rats which were used in this study.…”

MicroRNAs in toxic acute kidney injury: Systematic scoping review of the current status

“…We then performed a detailed evaluation of each title and abstract and 1343 were excluded due to being other forms of AKI, miRNA studied in cells and tissues, review articles, conference abstracts or editorial/commentary, and book chapters. Finally, 35 articles were eligible for full‐text screening and we selected 21 for final analysis 12‐32 . The application of the screening process and selection criteria is summarized in Figure 1.…”

“…The numbers add up to more than 21 as three studies examined both humans and animals 21,24,27 . and there were multiple agents studied in two publications 13,15 …”

“…We found only a few miRNAs that were shown multiple times to detect AKI after nephrotoxic agent exposure. Notably, miR‐218 was associated with cisplatin, gentamicin, and puromycin‐induced AKI 14,15,20,25,29,30 . MiR‐218 regulates cell migration via the Slit‐Robo pathway and promotes the motility of renal tubular epithelial cells that undergo epithelial mesenchymal transition 39 .…”