An Exploratory Evaluation of the Utility of Transcriptional and Urinary Kidney Injury Biomarkers for the Prediction of Aristolochic Acid–Induced Renal Injury in Male Rats

Abstract

The predictive value of different urinary and transcriptional biomarkers was evaluated in a proof-of-principle toxicology study in rats using aristolochic acid (AA), a known nephrotoxic agent. Male Wistar rats were orally dosed with 0.1, 1, or 10 mg/kg for 12 days. Urine was collected on days 1, 5, and 12 over 24 hours. Gene expression analysis was also conducted using quantitative realtime polymerase chain reaction and Illumina whole-genome chips. Protein biomarkers (Kim-1, Timp-1, vascular endothelial growth factor, osteopontin, clusterin, cystatin C, calbindin D-28K, b2-microglobulin, a-glutathione S-transferase, GSTY1b, RPA-1, and neutrophil gelatinase-associated lipocalin) were measured in these urine samples. Treatment with AA resulted in a slight dose-and/or time-dependent increase in urinary b2-microglobulin, lipocalin 2, and osteopontin before an increase in serum creatinine or serum urea nitrogen was observed. A strong decrease in urinary calbindin D-28K was also detected. The Compugen Ltd. prediction model scored both the 1-and 10-mg/kg AA dose groups as positive for nephrotoxicity despite the absence of renal histopathological changes. In addition, several previously described transcriptional biomarkers were identified as early predictors of renal toxicity as they were detected before morphological alterations had occurred. Altogether, these findings demonstrated the predictive values of renal biomarkers approved by the Food and Drug Administration, European Medicines Agency, and Pharmaceuticals & Medical Devices Agency in AA-induced renal injury in rats and confirmed the utility of renal transcriptional biomarkers for detecting progression of compound-induced renal injury in rats. In addition, several transcriptional biomarkers identified in this exploratory study could present early predictors of renal tubular epithelium injury in rats.Keywords aristolochic acid, nephrotoxicity biomarker, lipocalin 2/NGAL, osteopontin, prediction model, Nrf-2 signalingThe early detection of renal damage has been an important challenge in pharmaceutical drug development. Serum creatinine or serum urea nitrogen (SUN), the major clinicopathological markers currently employed to identify changes in kidney function, are not suitable for the early detection of acute kidney injury (AKI). Due to the functional reserve of the kidney, changes in these clinical markers appear late, when as much as two-thirds of the renal biomass has been lost. 33In the past few decades, intensive research has been conducted to identify additional more accurate biomarkers for clinical and preclinical safety assessment. 8,12 This whole process has been driven by the Predictive Safety Testing Consortium and the International Life Science Institute/Health and Environmental Sciences Institute. In the study reported here, our objective was to evaluate the predictive (observation of adverse events on a molecular basis before morphological and/or physiological alterations are established) performance of several novel urinary and transcription...

Effects of arginine vasopressin on the urine proteome in rats

“…Osteopontin can be used as a biomarker for ovarian cancer ( Rainczuk et al, 2013 ), kidney stones ( Bautista et al, 1996 ), bladder cancer ( Yang et al, 2011 ) and drug-induced kidney injury ( Phillips et al, 2016 ). Calbindin can serve as a biomarker for distal nephron segment injuries ( Iida et al, 2014 ), nephrotoxicity ( Hoffmann et al, 2010 ), acute kidney injury ( Togashi et al, 2012 ) and drug-induced renal injury ( Fuchs et al, 2014 ). Complement C3 can act as a biomarker of IgA nephropathy ( Liu et al, 2014 ) and glomerulonephritis ( Cumming et al, 1976 ).…”

The Impact of p53 on Aristolochic Acid I-Induced Gene Expression In Vivo

“…NGAL is also over-expressed in human cancers [45]. In terms of AAI treatment, previous in vivo work demonstrated that Ngal can be used as a biomarker of exposure [46]. Moreover, LCN2 was upregulated in AAI-exposed TP53(+/+) and TP53(-/-) HCT116 cells [28].…”

Peer Review #2 of "Effects of arginine vasopressin on the urine proteome in rats (v0.1)"

“…Osteopontin can be used as a biomarker for ovarian cancer (Rainczuk et al 2013), kidney stones (Bautista et al 1996), bladder cancer (Yang et al 2011) and drug-induced kidney injury (Phillips et al 2016). Calbindin can serve as a biomarker for distal nephron segment injuries (Iida et al 2014), nephrotoxicity (Hoffmann et al 2010), acute kidney injury (Togashi et al 2012)and drug-induced renal injury (Fuchs et al 2014).…”