Transporter-dependent cytotoxicity of antiviral drugs in primary cultures of human proximal tubular cells

The role of plasma membrane transporters in the nephrotoxicity of two antiretroviral drugs, cidofovir and tenofovir, was studied in primary cultures of human proximal tubular (hPT) cells. Cells were grown on Transwell filter inserts to maintain epithelial polarity and access to either the apical or basolateral plasma membrane. The function of relevant membrane transporters, organic anion transporter 1 and 3 (OAT1/3), P-glycoprotein (multidrug resistance protein-1; P-gp or MDR1), and organic cation transporter 2 (OCT2), was validated by measurements of apical-to-basolateral and basolateral-to-apical fluxes of furosemide, digoxin, and metformin, respectively. Acute cytotoxicity of cidofovir (0, 10, 50, 150, or 300 mu M) in the absence or presence of 500 mu M probenecid, tenofovir disoproxil fumarate (0, 20, 90, 180, or 360 mu M) in the absence or presence of 500 mu M probenecid, or cisplatin (0, 20, 90, 180, or 360 mu M) as a positive control in the absence or presence of 500 mu M cimetidine, was assessed after 4-h incubations by determinations of release of lactate dehydrogenase (LDH), gamma-glutamyltransferase (GGT), N-acetyl-beta-D-glucosaminidase (NAG), or Kidney Injury Molecule(-1) (KIM-1). Cell death generally agreed with each of the four biomarkers, was generally greater when cidofovir or tenofovir was added to the upper compartment, and was markedly diminished in the presence of the appropriate transport inhibitor. Additionally, the extent of cytotoxicity caused by the two antiviral drugs was similar to that caused by cisplatin. The results demonstrate the importance of plasma membrane transport of antiviral drugs to elicit cytotoxicity in the hPT cell.