Discovery of Genetic Variants of the Kinases That Activate Tenofovir in a Compartment-specific Manner.
Tenofovir (TFV) is used in combination with other antiretroviral drugs for human immunodeficiency virus (HIV) treatment and prevention. TFV requires two phosphorylation steps to become pharmacologically active; however, the kinases that activate TFV in cells and tissues susceptible to HIV infection have yet to be identified. Peripheral blood mononuclear cells (PBMC), vaginal, and colorectal tissues were transfected with siRNA targeting nucleotide kinases, incubated with TFV, and TFV-monophosphate (TFV-MP) and TFV-diphosphate (TFV-DP) were measured using mass spectrometry-liquid chromatography. Adenylate kinase 2 (AK2) performed the first TFV phosphorylation step in PBMC, vaginal, and colorectal tissues. Interestingly, both pyruvate kinase isozymes, muscle (PKM) or liver and red blood cell (PKLR), were able to phosphorylate TFV-MP to TFV-DP in PBMC and vaginal tissue, while creatine kinase, muscle (CKM) catalyzed this conversion in colorectal tissue. In addition, next-generation sequencing of the Microbicide Trials Network MTN-001 clinical samples detected 71 previously unreported genetic variants in the genes encoding these kinases. In conclusion, our results demonstrate that TFV is activated in a compartment-specific manner. Further, genetic variants have been identified that could negatively impact TFV activation, thereby compromising TFV efficacy in HIV treatment and prevention.
Department of Pharmacology & Molecular Sciences, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA ; Department of Medicine (Division of Clinical Pharmacology), Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA.
Julie M
JM
Lade
Department of Pharmacology & Molecular Sciences, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA ; Department of Medicine (Division of Clinical Pharmacology), Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA.
Elaine E
EE
To
Department of Pharmacology & Molecular Sciences, Johns Hopkins University School of Medicine, 600 North Wolfe Street, Blalock 569, Baltimore, MD 21287, USA ; Department of Medicine (Division of Clinical Pharmacology), Johns Hopkins University School of Medicine, 600 North Wolfe Street, Blalock 569, Baltimore, MD 21287, USA.
Craig W
CW
Hendrix
Department of Pharmacology & Molecular Sciences, Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA ; Department of Medicine (Division of Clinical Pharmacology), Johns Hopkins University School of Medicine, 725 North Wolfe Street, Biophysics 307, Baltimore, MD 21205, USA.
Namandjé N
NN
Bumpus
D000328:Adult; D019380:Anti-HIV Agents / Q000494:pharmacology / Q000627:therapeutic use; D002451:Cell Compartmentation; D003710:Demography; D005006:Ethnicity; D005260:Female; D055785:Gene Knockdown Techniques; D014644:Genetic Variation; D015658:HIV Infections / Q000188:drug therapy; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D008297:Male; D008875:Middle Aged; D020125:Mutation, Missense / Q000235:genetics; D011494:Protein Kinases / Q000235:genetics*; D034741:RNA, Small Interfering / Q000378:metabolism; D000068698:Tenofovir / Q000494:pharmacology* / Q000627:therapeutic use; D055815:Young Adult
D019380:Anti-HIV Agents; D034741:RNA, Small Interfering; D000068698:Tenofovir; D011494:Protein Kinases
AK2, adenylate kinase 2; CKM, creatine kinase, muscle; GUK1, guanylate kinase 1; HIV; HIV pre-exposure prophylaxis; HIV, human immunodeficiency virus; MTN-001, Microbicide Trials Network Study MTN-001; Microbicide Trials Network study MTN-001; NME1, NME/NM23 nucleoside diphosphate kinase 1; Nucleotide kinases; PBMC, peripheral blood mononuclear cells; PKLR, pyruvate kinase, liver and red blood cell; PKM, pyruvate kinase, muscle; PrEP, pre-exposure prophylaxis; SNV, single-nucleotide variant; TFV, tenofovir; TFV-DP, tenofovir-diphosphate; TFV-MP, tenofovir-monophosphate; Targeted next-generation sequencing; Tenofovir activation