Genetic Variation of the Kinases That Phosphorylate Tenofovir and Emtricitabine in Peripheral Blood Mononuclear Cells

Dominique B. Figueroa, Erin P. Madeen, Joseph Tillotson, Paul Richardson, Leslie Cottle, Marybeth McCauley, Raphael J. Landovitz, Adriana Andrade, Craig Hendrix, Kenneth H. Mayer, Timothy Wilkin, Roy M. Gulick, Namandje N Bumpus

Research output: Contribution to journalArticle

Abstract

Tenofovir (TFV) disoproxil fumarate and emtricitabine (FTC) are used in combination for HIV treatment and pre-exposure prophylaxis (PrEP). TFV disoproxil fumarate is a prodrug that undergoes diester hydrolysis to TFV. FTC and TFV are nucleoside/nucleotide reverse transcriptase inhibitors that upon phosphorylation to nucleotide triphosphate analogs competitively inhibit HIV reverse transcriptase. We previously demonstrated that adenylate kinase 2, pyruvate kinase, muscle and pyruvate kinase, liver and red blood cell phosphorylate TFV in peripheral blood mononuclear cells (PBMC). To identify the kinases that phosphorylate FTC in PBMC, siRNAs targeted toward kinases that phosphorylate compounds structurally similar to FTC were delivered to PBMC, followed by incubation with FTC and the application of a matrix-assisted laser desorption ionization-mass spectrometry method and ultra high performance liquid chromatography-UV to detect the formation of FTC phosphates. Knockdown of deoxycytidine kinase decreased the formation of FTC-monophosphate, while siRNA targeted toward thymidine kinase 1 decreased the abundance of FTC-diphosphate. Knockdown of either cytidine monophosphate kinase 1 or phosphoglycerate kinase 1 decreased the abundance of FTC-triphosphate. Next-generation sequencing of genomic DNA isolated from 498 HIV-uninfected participants in the HIV Prevention Trials Network 069/AIDS Clinical Trials Group A5305 clinical study, revealed 17 previously unreported genetic variants of TFV or FTC phosphorylating kinases. Of note, four individuals were identified as simultaneous carriers of variants of both TFV and FTC activating kinases. These results identify the specific kinases that activate FTC in PBMC, while also providing further insight into the potential for genetic variation to impact TFV and FTC activation.

Original languageEnglish (US)
Pages (from-to)421-429
Number of pages9
JournalAIDS Research and Human Retroviruses
Volume34
Issue number5
DOIs
StatePublished - May 1 2018

Fingerprint

Tenofovir
Blood Cells
Phosphotransferases
Pyruvate Kinase
HIV
Nucleotides
Deoxycytidine Kinase
Phosphoglycerate Kinase
HIV Reverse Transcriptase
Reverse Transcriptase Inhibitors
Emtricitabine
Diphosphates
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Prodrugs
DNA Sequence Analysis
Nucleosides

Keywords

  • emtricitabine
  • kinase
  • nucleotide reverse transcriptase inhibitor
  • pharmacogenetics
  • tenofovir

ASJC Scopus subject areas

  • Immunology
  • Virology
  • Infectious Diseases

Cite this

Genetic Variation of the Kinases That Phosphorylate Tenofovir and Emtricitabine in Peripheral Blood Mononuclear Cells. / Figueroa, Dominique B.; Madeen, Erin P.; Tillotson, Joseph; Richardson, Paul; Cottle, Leslie; McCauley, Marybeth; Landovitz, Raphael J.; Andrade, Adriana; Hendrix, Craig; Mayer, Kenneth H.; Wilkin, Timothy; Gulick, Roy M.; Bumpus, Namandje N.

In: AIDS Research and Human Retroviruses, Vol. 34, No. 5, 01.05.2018, p. 421-429.

Research output: Contribution to journalArticle

Figueroa, DB, Madeen, EP, Tillotson, J, Richardson, P, Cottle, L, McCauley, M, Landovitz, RJ, Andrade, A, Hendrix, C, Mayer, KH, Wilkin, T, Gulick, RM & Bumpus, NN 2018, 'Genetic Variation of the Kinases That Phosphorylate Tenofovir and Emtricitabine in Peripheral Blood Mononuclear Cells', AIDS Research and Human Retroviruses, vol. 34, no. 5, pp. 421-429. https://doi.org/10.1089/aid.2017.0243
Figueroa, Dominique B. ; Madeen, Erin P. ; Tillotson, Joseph ; Richardson, Paul ; Cottle, Leslie ; McCauley, Marybeth ; Landovitz, Raphael J. ; Andrade, Adriana ; Hendrix, Craig ; Mayer, Kenneth H. ; Wilkin, Timothy ; Gulick, Roy M. ; Bumpus, Namandje N. / Genetic Variation of the Kinases That Phosphorylate Tenofovir and Emtricitabine in Peripheral Blood Mononuclear Cells. In: AIDS Research and Human Retroviruses. 2018 ; Vol. 34, No. 5. pp. 421-429.
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