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MRP4: A previously unidentified factor in resistance to nucleoside-based antiviral drugs

Abstract

Dideoxynucleosides, which are potent inhibitors of HIV reverse transcriptase and other viral DNA polymerases, are a common component of highly active anti-retroviral therapy (HAART) (ref. 1). Six reverse transcriptase inhibitors have been approved for human use: azidothymidine; 2'3'-dideoxycytidine; 2'3'-dideoxyinosine; 2',3'-didehydro-3'deoxythymidine; 2',3'-dideoxy-3'-thiacytidine; and 4-[2-amino-6-(cyclopropylamino) -9H-purin-9-yl]-2-cyclopentene-1-methanol. Although drug-resistant HIV strains resulting from genetic mutation have emerged in patients treated with HAART (ref. 1), some patients show signs of drug resistance in the absence of drug-resistant viruses2,3. In our study of alternative or additional mechanisms of resistance operating during antiviral therapy, overexpression and amplification of the MRP4 gene correlated with ATP-dependent efflux of PMEA (9-(2-phosphonylmethoxyethyl)adenine) and azidothymidine monophosphate from cells and, thus, with resistance to these drugs. Overexpression of MRP4 mRNA and MRP4 protein severely impaired the antiviral efficacy of PMEA, azidothymidine and other nucleoside analogs. Increased resistance to PMEA and amplification of the MRP4 gene correlated with enhanced drug efflux; transfer of chromosome 13 containing the amplified MRP4 gene conferred resistance to PMEA. MRP4 is the first transporter, to our knowledge, directly linked to the efflux of nucleoside monophosphate analogs from mammalian cells.

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Figure 1: Selective MRP4 gene amplification in PMEA-resistant cells.
Figure 2: Amplification and overexpression of MRP4 in CEMss cells selected for PMEA resistance.

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Acknowledgements

We thank W. Evans and E. Tuomanen and members of the Schuetz laboratory for their comments; A. Gandhi and A. Tatum for their technical assistance; F. Witte for scientific editing; and T. Look and V. Valentine for the FISH studies. This work was supported by NIH Grants ES/GM 5851 and 8568, AI27652, and CA21765 and by the American Lebanese Syrian Associated Charities (ALSAC).

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Correspondence to John D. Schuetz.

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Schuetz, J., Connelly, M., Sun, D. et al. MRP4: A previously unidentified factor in resistance to nucleoside-based antiviral drugs. Nat Med 5, 1048–1051 (1999). https://doi.org/10.1038/12487

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