REverSe TRanscrIptase chain termination (RESTRICT) for selective measurement of nucleotide analogs used in HIV care and prevention

Ayokunle O Olanrewaju; Benjamin P Sullivan; Alicia H Gim; Cosette A Craig; Derin Sevenler; Andrew T Bender; Paul K Drain; Jonathan D Posner

doi:10.1002/btm2.10369

REverSe TRanscrIptase chain termination (RESTRICT) for selective measurement of nucleotide analogs used in HIV care and prevention

Bioeng Transl Med. 2022 Jul 14;8(1):e10369. doi: 10.1002/btm2.10369. eCollection 2023 Jan.

Authors

Ayokunle O Olanrewaju^{1

2}, Benjamin P Sullivan¹, Alicia H Gim³, Cosette A Craig¹, Derin Sevenler⁴, Andrew T Bender¹, Paul K Drain^{5

6

7}, Jonathan D Posner^{1

3

8}

Affiliations

¹ Department of Mechanical Engineering University of Washington Seattle Washington USA.
² Department of Bioengineering University of Washington Seattle Washington USA.
³ Department of Chemical Engineering University of Washington Seattle Washington USA.
⁴ Center for Engineering in Medicine and Surgery Massachusetts General Hospital Boston Massachusetts USA.
⁵ Department of Epidemiology University of Washington Seattle Washington USA.
⁶ Department of Global Health University of Washington Seattle Washington USA.
⁷ Department of Medicine University of Washington Seattle Washington USA.
⁸ Department of Family Medicine University of Washington Seattle Washington USA.

Abstract

Sufficient drug concentrations are required for efficacy of antiretroviral drugs used in HIV care and prevention. Measurement of nucleotide analogs, included in most HIV medication regimens, enables monitoring of short- and long-term adherence and the risk of treatment failure. The REverSe TRanscrIptase Chain Termination (RESTRICT) assay rapidly infers the concentration of intracellular nucleotide analogs based on the inhibition of DNA synthesis by HIV reverse transcriptase enzyme. Here, we introduce a probabilistic model for RESTRICT and demonstrate selective measurement of multiple nucleotide analogs using DNA templates designed according to the chemical structure of each drug. We measure clinically relevant concentrations of tenofovir diphosphate, emtricitabine triphosphate, lamivudine triphosphate, and azidothymidine triphosphate with agreement between experiment and theory. RESTRICT represents a new class of activity-based assays for therapeutic drug monitoring in HIV care and could be extended to other diseases treated with nucleotide analogs.

Keywords: adherence monitoring; enzymatic assay; nucleotide analog; nucleotide reverse transcriptase inhibitor; reverse transcriptase inhibitor; tenofovir diphosphate; therapeutic drug monitoring.

Abstract

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