Absolute Measurement of Cardiac Injury-Induced microRNAs in Biofluids across Multiple Test Sites.

Autor:	Thompson KL; Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland 20993., Boitier E; Sanofi R&D, Disposition Safety and Animal Research, Vitry-Sur-Seine, France., Chen T; Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arizona 72079., Couttet P; Novartis Pharma AG, Basel, CH 4057, Switzerland., Ellinger-Ziegelbauer H; Toxicology, Bayer Pharma, Wuppertal, AG 42096, Germany., Goetschy M; Novartis Pharma AG, Basel, CH 4057, Switzerland., Guillemain G; Novartis Pharma AG, Basel, CH 4057, Switzerland., Kanki M; Astellas Pharma Inc, Osaka 532-8514, Japan., Kelsall J; AstraZeneca Ltd, Alderley Park, Macclesfield, Cheshire SK10 4TG, UK., Mariet C; Sanofi R&D, Disposition Safety and Animal Research, Vitry-Sur-Seine, France., de La Moureyre-Spire C; Institut De Recherches Servier, 78290 Croissy Sur Seine, France., Mouritzen P; Exiqon, Vedbaek DK-2950, Denmark., Nassirpour R; Pfizer, Andover, Massachusetts 01810., O'Lone R; ILSI Health and Environmental Sciences Institute, Washington, DC 20005 Rolone@hesiglobal.org., Pine PS; National Institute of Standards and Technology, Stanford, California 94305., Rosenzweig BA; Center for Drug Evaluation and Research, Food and Drug Administration, Silver Spring, Maryland 20993., Sharapova T; AbbVie, Abbott Park, Illinois 60064., Smith A; Eli Lilly, Indianapolis, Indiana 46285., Uchiyama H; Takeda Pharmaceutical Co Ltd, Fujisawa, Kanagawa 251-8555, Japan., Yan J; Division of Genetic and Molecular Toxicology, National Center for Toxicological Research, Food and Drug Administration, Jefferson, Arizona 72079., Yuen PS; NIH/NIDDK, Bethesda, Maryland 20892., Wolfinger R; SAS Institute Inc, Cary, North Carolina 27513.
Jazyk:	angličtina
Zdroj:	Toxicological sciences : an official journal of the Society of Toxicology [Toxicol Sci] 2016 Nov; Vol. 154 (1), pp. 115-125. Date of Electronic Publication: 2016 Sep 07.
DOI:	10.1093/toxsci/kfw143
Abstrakt:	Extracellular microRNAs (miRNAs) represent a promising new source of toxicity biomarkers that are sensitive indicators of site of tissue injury. In order to establish reliable approaches for use in biomarker validation studies, the HESI technical committee on genomics initiated a multi-site study to assess sources of variance associated with quantitating levels of cardiac injury induced miRNAs in biofluids using RT-qPCR. Samples were generated at a central site using a model of acute cardiac injury induced in male Wistar rats by 0.5 mg/kg isoproterenol. Biofluid samples were sent to 11 sites for measurement of 3 cardiac enriched miRNAs (miR-1-3p, miR-208a-3p, and miR-499-5p) and 1 miRNA abundant in blood (miR-16-5p) or urine (miR-192-5p) by absolute quantification using calibration curves of synthetic miRNAs. The samples included serum and plasma prepared from blood collected at 4 h, urine collected from 6 to 24 h, and plasma prepared from blood collected at 24 h post subcutaneous injection. A 3 parameter logistic model was utilized to fit the calibration curve data and estimate levels of miRNAs in biofluid samples by inverse prediction. Most sites observed increased circulating levels of miR-1-3p and miR-208a-3p at 4 and 24 h after isoproterenol treatment, with no difference seen between serum and plasma. The biological differences in miRNA levels and sample type dominated as sources of variance, along with outlying performance by a few sites. The standard protocol established in this study was successfully implemented across multiple sites and provides a benchmark method for further improvements in quantitative assays for circulating miRNAs. (Published by Oxford University Press on behalf of the Society of Toxicology 2016. This work is written by US Government employees and is in the public domain in the US.)
Databáze:	MEDLINE
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