Ifosfamide toxicity in cultured proximal renal tubule cells

Abstract

Renal injury is a common side effect of the chemotherapeutic agent ifosfamide. Current evidence suggests that ifosfamide metabolites, particularly chloroacetaldehyde, produced within the kidney contribute to nephrotoxicity. The present study examined the effects of ifosfamide and its metabolites, chloroacetaldehyde and acrolein, on rabbit proximal renal tubule cells in primary culture, using a transwell culture system that allows separate access to apical and basolateral cell surfaces. The ability of the uroprotectant medications sodium 2-mercaptoethanesulfonate (mesna) and amifostine to prevent chloroacetaldehyde-and acrolein-induced renal cell injury was also assessed. Ifosfamide (2,000-4,000 microM) did not affect transcellular inulin diffusion but caused a modest but significant impairment in organic ion transport; this impairment was greater when ifosfamide was added to the basolateral compartment of the transwell. Chloroacetaldehyde and acrolein (6.25-100 microM) produced dose-dependent impairments in transcellular inulin diffusion and organic ion transport. Chloroacetaldehyde was a more potent toxin than acrolein. Co-administration of mesna or amifostine prevented metabolite toxicity. Amifostine was only protective when added to the apical compartment of transwells. These results show that ifosfamide is taken up by renal tubule cells preferentially through their basolateral surfaces, and supports the hypothesis that chloroacetaldehyde is primarily responsible for ifosfamide-induced nephrotoxicity. The protective effect of mesna and amifostine in vitro contrasts with clinical experience showing that these medications do not eliminate ifosfamide nephrotoxicity in vivo.

In vivo mesna and amifostine do not prevent chloroacetaldehyde nephrotoxicity in vitro

“…This is why, in our study, the long-term toxicity of very low concentrations of CAA was studied in precision-cut cortical slices. In the sham group, our observation that the 24-h incubation of kidney slices with low concentration of CAA led to significant decreases in cellular levels of ATP, total glutathione and coenzyme A + acetyl-coenzyme A is in accord with previously published studies in cultured cells [30,32,33,42,43]. In contrast to in vitro addition, in vivo mesna or amifostine administration did not prevent CAA toxicity.…”

“…We also demonstrated that the nephrotoxic effects of 0.5 mM CAA were prevented by addition to the incubation medium of an equimolar concentration of mesna or a tenfold higher concentration of amifostine [12]. These results were confirmed by several studies conducted by other authors, showing that mesna and amifostine could prevent toxicity of CAA in vitro [30][31][32][33]. Despite the fact that the protecting concentration of thiols compounds were quite different among the various studies (from 200 to 800 μM), it must be emphasised that the protecting effect obtained under in vitro conditions was very efficacious and was promising to also prevent CAA toxicity in clinical practice.…”

Phylogenetic Fate Mapping: Theoretical and Experimental Studies Applied to the Development of Mouse Fibroblasts

“…Here we demonstrate an alternative to wholegenome amplification, in which we have cultured single fibroblasts briefly as clonal isolates for a few generations to extract enough genomic DNA to complete multiple PCR genotyping reactions. Many types of cells, including hepatocytes (Tateno and Yoshizato 1999), adipocytes (Sugihara et al 1987), astrocytes (Mbarek et al 1998), renal tubular cells (Springate and Taub 2007), and epithelial cells from colon (Follmann et al 2000), prostate (McKeehan et al 1984), and mammary glands (Soule and McGrath 1986) can grow in primary cell culture, if not immortally, then for at least a few generations. And for those cell types that may be terminally differentiated or otherwise incapable of reentry into the cell cycle, another possibility involves expression of a conditionally immortalizing gene from a DNA tumor virus, such as the SV40 T-antigen.…”

Ifosfamide nephrotoxicity in children: a mechanistic base for pharmacological prevention

“…Woodland et al [14] then hypothesized that local renal metabolism of IFO to CAA -not systemic exposure -produces renal tubular damage. Although other toxic species such as acrolein are produced from IFO, metabolism studies by Springate and Taub [34] have demonstrated CAA to be the main nephrotoxic IFO metabolite.…”