Evidence map›Paper›PMID 27557296›Full record

ArticleBiomedical chromatography : BMC2017

Development and validation of an LC-MS/MS quantitative method for endogenous deoxynucleoside triphosphates in cellular lysate.

Xinhui Chen, Kevin J McAllister, Brandon Klein, Lane R Bushman, Peter L Anderson

Abstract read
In one paragraph

Article in Biomedical chromatography : BMC, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 2 papers.

0numbers the graph read from it
0cells of the map it votes in
2citing papers in PubMed
–field-weighted citation impact
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

2 citing papers in PubMed.

  1. Article
  2. Observational
4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

5 authors.

Xinhui ChenUniversity of Colorado, Skaggs School of Pharmacy and Pharmaceutical Sciences, Aurora, CO, USA.
Kevin J McAllisterUniversity of Colorado, School of Medicine, Aurora, CO, USA.
Brandon KleinUniversity of Colorado, Skaggs School of Pharmacy and Pharmaceutical Sciences, Aurora, CO, USA.
Lane R BushmanUniversity of Colorado, Skaggs School of Pharmacy and Pharmaceutical Sciences, Aurora, CO, USA.
Peter L AndersonUniversity of Colorado, Skaggs School of Pharmacy and Pharmaceutical Sciences, Aurora, CO, USA.

Funding

Colorado Clinical and Translational Sciences InstituteUL1TR001082 · NCATS · UNIVERSITY OF COLORADO DENVER · PI SOKOL, RONALD J. · 2013 to 2017
$48.0M
Cellular pharmacology of tenofovir and emtricitabine for HIV prophylaxisU01AI084735 · NIAID · UNIVERSITY OF COLORADO DENVER · PI ANDERSON, PETER L. · 2009 to 2012
$4.2M
PrEP and ART adherence monitoring using dried blood spotsU01AI106499 · NIAID · UNIVERSITY OF COLORADO DENVER · PI ANDERSON, PETER L. · 2013 to 2016
$2.9M
Colorado Clinical and Translational Sciences InstituteTL1TR001081 · NCATS · UNIVERSITY OF COLORADO DENVER · PI SOKOL, RONALD J. · 2013 to 2017
$1.3M
NCATS NIH HHS TL1 TR001081NCATS NIH HHS UL1 TR001082NIAID NIH HHS U01 AI084735NIAID NIH HHS U01 AI106499
6 · The paper itself

Abstract

The endogenous deoxynucleoside triphosphate (dNTP) pool includes deoxyadenosine triphosphate (dATP), deoxycytidine triphosphate (dCTP), deoxyguanosine triphosphate (dGTP) and thymidine triphosphate (TTP). The endogenous dNTP pool is regulated by complex enzymatic pathways that can be targeted by drugs, such as antimetabolites. In addition, these components compete with antiviral nucleos(t)ide analog triphosphates, contributing to the mechanism of pharmacologic action. This communication describes the development and validation of a sensitive method to quantify the intracellular dNTP pool in cellular lysates. The extraction process was optimized for dNTPs using an indirect strategy - the separation of mono-, di- and triphosphate moieties by strong anion exchange, dephosphorylation of target fractions to molar equivalent nucleosides - followed by sensitive quantitation using liquid chromatography-tandem mass spectrometry. The validated analytical range was 50-2500 fmol/sample for each dNTP. The assay was used to quantify dNTPs in peripheral blood mononuclear cells from 40 clinical research participants (n = 279 samples). Median (interquartile range) concentrations were 143 (116, 169) for dATP, 737 (605, 887) for dCTP, 237 (200, 290) for dGTP and 315 (220, 456) for TTP, in femtomoles per million cells. This method allows for future studies of endogenous dNTP disposition in subjects receiving antimetabolites or nucleos(t)ide analogs, or for other clinical scenarios.

Indexed as

Chromatography, LiquidDeoxyadenine NucleotidesDeoxycytosine NucleotidesTandem Mass SpectrometryThymine Nucleotides2'-deoxyadenosine triphosphate2'-deoxycytidine 5'-triphosphateDeoxyadenine NucleotidesDeoxycytosine Nucleotidesthymidine 5'-triphosphateThymine Nucleotidesendogenous dNTPintracellularLC-MS/MSpharmacodynamicspharmacokinetics

Identifiers

PMID27557296
PMCPMC5293618

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Registered trials

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.