Identification of Noninvasive Biomarkers for Nephrotoxicity Using HK-2 Human Kidney Epithelial Cells

Abstract

The kidney is an important site of xenobiotic-induced toxicity. Because the traditional markers of renal injury indicate only severe renal damage, new biomarkers are needed for a more sensitive and reliable evaluation of renal toxicity. This study was designed to identify in vitro noninvasive biomarkers for efficient assessment of nephrotoxicity by using cisplatin as a model of nephrotoxic compounds. To this end, a comparative proteomic analysis of conditioned media from HK-2 human kidney epithelial cells treated with cisplatin was performed. Here, we identified pyruvate kinase M1/M2 isoform M2 (PKM2) and eukaryotic translation elongation factor 1 gamma (EF-1γ) as potential biomarker candidates for evaluation of nephrotoxicity. PKM2 and EF-1γ were increased by cisplatin in a kidney cell-specific manner, most likely due to cisplatin-induced apoptosis. The increase of PKM2 and EF-1γ levels in conditioned media was also observed in the presence of other nephrotoxic agents with different cytotoxic mechanisms such as CdCl2, HgCl2, and cyclosporine A. Rats treated with cisplatin, CdCl2, or HgCl2 presented increased levels of PKM2 and EF-1γ in the urine and kidney tissue. Taken together, this study identified two noninvasive biomarker candidates, PKM2 and EF-1γ, by comparative proteomic analysis. These new biomarkers may offer an alternative to traditional renal markers for efficient evaluation of nephrotoxicity.

Mechanism of Cisplatin-Induced Cytotoxicity Is Correlated to Impaired Metabolism Due to Mitochondrial ROS Generation

“…Each animal was kept in a rat metabolic cage overnight and 24-h urine samples were collected in the morning. Animals were monitored daily by the disease activity index that includes loss of body weight, serum levels of blood urea nitrogen (BUN) and creatinine, and hair loss as described previously [ 24 – 26 ]. Animals were euthanized under anesthesia by cervical dislocation when they lost more than 20% of starting bodyweight.…”

An integrative view of cisplatin-induced renal and cardiac toxicities: Molecular mechanisms, current treatment challenges and potential protective measures

“…Further, cisplatin treatment significantly increased mRNA levels of KIM-1, TIMP-1 and calbindin, suggesting involvement of transcriptional activation (Sohn et al 2013). Other authors also suggested pyruvate kinase M2 (PKM2) and eukaryotic translation elongation factor 1 gamma (EF-1γ) as biomarker candidates for detection and evaluation of cisplatin-induced renal toxicity and nephrotoxicity in general (Kim et al, 2014). They reported high levels of PKM2 and EF-1γ in conditioned media of HK-2 cells and in the urine and kidney tissue of rats upon 24 hours and 72 hours of cisplatin administration (Kim et al, 2014).…”

“…Other authors also suggested pyruvate kinase M2 (PKM2) and eukaryotic translation elongation factor 1 gamma (EF-1γ) as biomarker candidates for detection and evaluation of cisplatin-induced renal toxicity and nephrotoxicity in general (Kim et al, 2014). They reported high levels of PKM2 and EF-1γ in conditioned media of HK-2 cells and in the urine and kidney tissue of rats upon 24 hours and 72 hours of cisplatin administration (Kim et al, 2014). These may not have been the best study designs, as HK-2 cells are cancer cells and are not contact inhibited and thus keep proliferating.…”

Proteomic analysis of rat kidney under maleic acid treatment by SWATH‐MS technology

“…The differentially expressed proteins were involved in three related pathways: the glycolysis/gluconeogenesis, fructose and mannose metabolism, and pentose phosphate pathways. In previous studies, both Song et al and Kim et al pointed out that damage to the kidneys would inhibit glycolytic effects . Basile et al further demonstrated that the inhibition of glycolysis leads to apoptosis and cell necrosis .…”