Kia Richard, Kelly Lorna, Sison-Young Rowena L C, Zhang Fang, Pridgeon Chris S, Heslop James A, Metcalfe Pete, Kitteringham Neil R, Baxter Melissa, Harrison Sean, Hanley Neil A, Burke Zoë D, Storm Mike P, Welham Melanie J, Tosh David, Küppers-Munther Barbara, Edsbagge Josefina, Starkey Lewis Philip J, Bonner Frank, Harpur Ernie, Sidaway James, Bowes Joanne, Fenwick Stephen W, Malik Hassan, Goldring Chris E P, Park B Kevin
MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK Stem Cells for Safer Medicines, 7th Floor, Southside, 105 Victoria Street, London SW1E 6QT, UK Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK School of Medicine and Dentistry, University of Central Lancashire, Preston PR1 2HE, UK Centre for Endocrinology and Diabetes, Institute of Human Development, Faculty of Medical and Human Sciences, Manchester Academic Health Science Centre, University of Manchester, Manchester M13 9PT, UK Endocrinology Department, Central Manchester University Hospitals NHS Foundation Trust, Oxford Road, Manchester M13 9PT, UK Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK, Takara Bio Europe AB (former Cellartis), Arvid Wallgrens Backe 20, 413 46 Göteborg, Sweden, School of Life Sciences, The Systems Biology Research Centre, University of Skövde, Box 408, 541 28 Skövde, Sweden, MRC Centre for Regenerative Medicine, SCRM Building, The University of Edinburgh, Edinburgh Bioquarter, 5 Little France Drive, Edinburgh EH16 4UU, UK, Newcastle University, Institute of Cellular Medicine, The Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK Drug Safety and Metabolism, AstraZeneca R&D, Alderley Park, Cheshire SK10 4TG, UK and North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK.
MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK Stem Cells for Safer Medicines, 7th Floor, Southside, 105 Victoria Street, London SW1E 6QT, UK Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK School of Medicine and Dentistry, University of Central Lancashire, Preston PR1 2HE, UK Centre for Endocrinology and Diabetes, Institute of Human Development, Faculty of Medical and Human Sciences, Manchester Academic Health Science Centre, University of Manchester, Manchester M13 9PT, UK Endocrinology Department, Central Manchester University Hospitals NHS Foundation Trust, Oxford Road, Manchester M13 9PT, UK Centre for Regenerative Medicine, Department of Biology and Biochemistry, University of Bath, Bath BA2 7AY, UK, Takara Bio Europe AB (former Cellartis), Arvid Wallgrens Backe 20, 413 46 Göteborg, Sweden, School of Life Sciences, The Systems Biology Research Centre, University of Skövde, Box 408, 541 28 Skövde, Sweden, MRC Centre for Regenerative Medicine, SCRM Building, The University of Edinburgh, Edinburgh Bioquarter, 5 Little France Drive, Edinburgh EH16 4UU, UK, Newcastle University, Institute of Cellular Medicine, The Medical School, Framlington Place, Newcastle upon Tyne NE2 4HH, UK Drug Safety and Metabolism, AstraZeneca R&D, Alderley Park, Cheshire SK10 4TG, UK and North Western Hepatobiliary Unit, Aintree University Hospital NHS Foundation Trust, Longmoor Lane, Liverpool L9 7AL, UK MRC Centre for Drug Safety Science, Department of Molecular and Clinical Pharmacology, Sherrington Buildings, Ashton Street, University of Liverpool, Liverpool L69 3GE, UK Stem Cells for Safer Medicines, 7th Floor, Southside, 105 Victoria Street, London SW1E 6QT, UK Faculty of Life Sciences, University of Manchester, Manchester M13 9PL, UK School of Medicine and Dentistry, University of Central Lancashire, Preston PR1 2HE, UK Centre fo
Toxicol Sci. 2015 Mar;144(1):173-85. doi: 10.1093/toxsci/kfu269. Epub 2014 Dec 18.
Emerging hepatic models for the study of drug-induced toxicity include pluripotent stem cell-derived hepatocyte-like cells (HLCs) and complex hepatocyte-non-parenchymal cellular coculture to mimic the complex multicellular interactions that recapitulate the niche environment in the human liver. However, a specific marker of hepatocyte perturbation, required to discriminate hepatocyte damage from non-specific cellular toxicity contributed by non-hepatocyte cell types or immature differentiated cells is currently lacking, as the cytotoxicity assays routinely used in in vitro toxicology research depend on intracellular molecules which are ubiquitously present in all eukaryotic cell types. In this study, we demonstrate that microRNA-122 (miR-122) detection in cell culture media can be used as a hepatocyte-enriched in vitro marker of drug-induced toxicity in homogeneous cultures of hepatic cells, and a cell-specific marker of toxicity of hepatic cells in heterogeneous cultures such as HLCs generated from various differentiation protocols and pluripotent stem cell lines, where conventional cytotoxicity assays using generic cellular markers may not be appropriate. We show that the sensitivity of the miR-122 cytotoxicity assay is similar to conventional assays that measure lactate dehydrogenase activity and intracellular adenosine triphosphate when applied in hepatic models with high levels of intracellular miR-122, and can be multiplexed with other assays. MiR-122 as a biomarker also has the potential to bridge results in in vitro experiments to in vivo animal models and human samples using the same assay, and to link findings from clinical studies in determining the relevance of in vitro models being developed for the study of drug-induced liver injury.
用于研究药物诱导毒性的新兴肝脏模型包括多能干细胞衍生的肝样细胞（HLCs）以及复杂的肝细胞与非实质细胞共培养体系，以模拟复杂的多细胞相互作用，重现人类肝脏中的生态位环境。然而，目前缺乏一种肝细胞扰动的特异性标志物，无法区分肝细胞损伤与非肝细胞类型或未成熟分化细胞造成的非特异性细胞毒性，因为体外毒理学研究中常规使用的细胞毒性检测依赖于所有真核细胞类型中普遍存在的细胞内分子。在本研究中，我们证明在细胞培养基中检测微小RNA-122（miR-122）可作为肝细胞富集的体外标志物，用于检测肝细胞同质培养物中药物诱导的毒性，以及异质培养物（如通过各种分化方案和多能干细胞系产生的HLCs）中肝细胞毒性的细胞特异性标志物，在这些异质培养物中，使用通用细胞标志物的传统细胞毒性检测可能并不适用。我们表明，当应用于细胞内miR-122水平较高的肝脏模型时，miR-122细胞毒性检测的灵敏度与测量乳酸脱氢酶活性和细胞内三磷酸腺苷的传统检测相似，并且可以与其他检测方法进行多重检测。MiR-122作为一种生物标志物，还有潜力通过相同的检测方法将体外实验结果与体内动物模型和人类样本联系起来，并在确定为药物性肝损伤研究而开发的体外模型的相关性时，将临床研究结果联系起来。
