Cytotoxicity of antiviral nucleotides adefovir and cidofovir is induced by the expression of human renal organic anion transporter 1.
The transport of organic anions in proximal convoluted tubules plays an essential role in the active secretion of a variety of small molecules by the kidney. In addition to other anionic substrates, the human renal organic anion transporter 1 (hOATI) is capable of transporting the nucleotide analogs adefovir and cidofovir. To investigate the involvement of hOATI in the mechanism of nephrotoxicity associated with these two clinically important antiviral agents, Chinese hamster ovary (CHO) cells were stably transfected with hOATI cDNA. The resulting CHOhOAT cells showed probenecid-sensitive and pH-dependent uptake of p-aminohippurate (Km = 15.4 FtM, V,,, ..ax = 20.6 pmol/106 cells min), a prototypical organic anion substrate. In addition, the stably expressed hOATI mediated efficient transport of adefovir (Km, = 23.8 tLM, V, a,, = 46.0 pmol/106 cells min) and cidofovir (K, = 58.0 /iM, Vt,ax = 103 pmol/106 cells * min) such that the levels of intracellular metabolites of both nucleotides were > 1 00-fold higher in CHOh OAT cells than in parental CHO. Consequently, adefovir and cidofovir were approximately 500-fold and 400-fold more cytotoxic, respectively, in CHOh OAT cells compared to CHO. The cytotoxicity of both drugs in CHOh OAT cells was markedly reduced in the presence of hOATI inhibitors. The cyclic prodrug of cidofovir, which exhibits reduced in vivo nephrotoxicity, was a poor substrate for hOATI and showed only marginally increased cytotoxicity in CHOh OAT cells. In conclusion, these studies demonstrate that hOATI plays a critical role in the organ-specific toxicity of adefovir and cidofovir, and indicates that CHOh OAT cells may represent a useful in vitro model to investigate the potential nephrotoxicity of clinically relevant organic anion agents.
Ho
Edmund S
ES
Gilead Sciences, Foster City, California.
Lin
Deborah C
DC
Gilead Sciences, Foster City, California.
Mendel
Dirk B
DB
Gilead Sciences, Foster City, California.
Cihlar
Tomas
T
Gilead Sciences, Foster City, California.
D000225:Adenine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000493:pharmacokinetics / Q000506:poisoning; D000409:Alanine / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D027321:Anion Transport Proteins; D000998:Antiviral Agents / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000493:pharmacokinetics / Q000506:poisoning*; D001692:Biological Transport / Q000502:physiology; D016466:CHO Cells; D002352:Carrier Proteins / Q000378:metabolism / Q000502:physiology*; D000077404:Cidofovir; D006224:Cricetinae; D003596:Cytosine / Q000031:analogs & derivatives* / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000493:pharmacokinetics / Q000506:poisoning; D006801:Humans; D007668:Kidney / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D062025:Organophosphate Poisoning; D063065:Organophosphonates; D009943:Organophosphorus Compounds / Q000037:antagonists & inhibitors / Q000378:metabolism / Q000493:pharmacokinetics; D011339:Probenecid / Q000494:pharmacology
D027321:Anion Transport Proteins; D000998:Antiviral Agents; D002352:Carrier Proteins; D063065:Organophosphonates; D009943:Organophosphorus Compounds; C047207:N-benzoylalanine; C053001:adefovir; D003596:Cytosine; D000225:Adenine; D000077404:Cidofovir; D000409:Alanine; D011339:Probenecid