Toxicity of ifosfamide and its metabolite chloroacetaldehyde in cultured renal tubule cells

Abstract

Renal injury is a common side effect of the chemotherapeutic agent ifosfamide. Current evidence suggests that the ifosfamide metabolite chloroacetaldehyde may contribute to this nephrotoxicity. The present study examined the effects of ifosfamide and chloroacetaldehyde on rabbit proximal renal tubule cells in primary culture. The ability of the uroprotectant medication sodium 2-mercaptoethanesulfonate (mesna) to prevent chloroacetaldehyde-induced renal cell injury was also assessed. Chloroacetaldehyde (12.5-150 microM) produced dose-dependent declines in neutral red dye uptake, ATP levels, glutathione content, and cell growth. Coadministration of mesna prevented chloroacetaldehyde toxicity while pretreatment of cells with the glutathione-depleting agent buthionine sulfoximine enhanced the toxicity of chloroacetaldehyde. Ifosfamide (1000-10,000 microM) toxicity was detected only at concentrations of 4000 microM or greater. Analysis of media collected from ifosfamide-treated cell cultures revealed the presence of several ifosfamide metabolites, demonstrating that renal proximal tubule cells are capable of biotransforming this chemotherapeutic agent. This primary renal cell culture system should prove useful in studying the cause and prevention of ifosfamide nephrotoxicity.

Alkylation of Adenosine Deaminase and Thioredoxin by Acrylamide in Human Cell Cultures

“…The concentrations that caused serious cytotoxicity (2 -4 mM) were comparable to those of other alkylating reagents tested in cultured human and rat hepatocytes (Boot, 1996). They were also similar to the IC 50 value of ifosfamide (a mustard alkylating agent) in rabbit proximal renal tubule cells (Springate et al, 1999). Hence, our data confi rms that acrylamide causes cytotoxicity effects in cell cultures and this cytotoxicity is most likely mediated by protein alkylation.…”

Mesna or cysteine prevents chloroacetaldehyde-induced cell death of human proximal tubule cells

“…However, chemotherapy using IFO is limited by severe cytotoxic side effects, including hemorrhagic cystitis and nephrotoxicity [3,4]. This is due to formation of the metabolites acrolein and chloroacetaldehyde (CAA) and possibly by ifosforamide mustard [5,6].…”

CONTRIBUTION OF CYP3A5 TO HEPATIC AND RENAL IFOSFAMIDEN-DECHLOROETHYLATION

“…Ifosfamide itself is not cytotoxic in cultured renal epithelial cells or isolated perfused rat kidneys exposed to ifosfamide concentrations that have been observed in cancer patients receiving ifosfamide (i.e., 160 -650 M) (Kurowski and Wagner, 1993;Mohrmann et al, 1993;Zamlauski-Tucker et al, 1994;Springate et al, 1999;Boddy and Yule, 2000). This raises the possibility that ifosfamide metabolites are nephrotoxic.…”

“…Both acrolein and chloroacetaldehyde cause toxicity in LLC-PK1 cells (Mohrmann et al, 1992(Mohrmann et al, , 1993; however, acrolein does not impair the function of isolated perfused rat kidneys or after long-term exposure in animal models (Parent et al, 1992;Zamlauski-Tucker et al, 1994). Chloroacetaldehyde has consistently shown a concentration-dependent cytotoxic effect in several in vitro models (i.e., porcine or rabbit cultured renal tubules and isolated perfused rat kidneys) with a minimum toxic concentration that ranges from 12.5 to 64 M (Mohrmann et al, 1993;Springate, 1997;Springate et al, 1999). In cultured primary human proximal tubules cells, the minimum chloroacetaldehyde concentration for toxicity was reported to be 500 M, which is well above the observed peak plasma concentrations (i.e., 0.5-109 M) in patients receiving standard doses of ifosfamide (Goren et al, 1986;Kurowski and Wagner, 1993;Dubourg et al, 2001).…”