In vitro/in silico prediction of drug induced steatosis in relation to oral doses and blood concentrations by the Nile Red assay

Fecha de publicación: 01/09/2022 Fecha Ahead of Print: 10/08/2022

Autores de IIS La Fe

Laia Tolosa Pardo

Autor
José Vicente Castell Ripoll

Autor

Participantes ajenos a IIS La Fe

Brecklinghaus, Tim
Albrecht, Wiebke
Duda, Julia
Kappenberg, Franziska
Gruendler, Lisa
Edlund, Karolina
Marchan, Rosemarie
Ghallab, Ahmed
Cadenas, Cristina
Rieck, Adrian
Vartak, Nachiket
Gardner, Iain
Halilbasic, Emina
Trauner, Michael
Ullrich, Anett
Zeigerer, Anja
Turgunbayer, Oezlem Demirci
Damm, Georg
Seehofer, Daniel
Rahnenfuhrer, Joerg
Hengstler, Jan G.

Grupos

Hepatología experimental y Trasplante hepático

Investigador Principal

Laia Tolosa Pardo
Unidad Mixta de Investigación en Hepatología Experimental

Investigador Principal

Ramiro Jover Atienza

Abstract

The accumulation of lipid droplets in hepatocytes is a key feature of drug-induced liver injury (DILI) and can be induced by a subset of hepatotoxic compounds. In the present study, we optimized and evaluated an in vitro technique based on the fluorescent dye Nile Red, further named Nile Red assay to quantify lipid droplets induced by the exposure to chemicals. The Nile Red assay and a cytotoxicity test (CTB assay) were then performed on cells exposed concentration-dependently to 60 different compounds. Of these, 31 were known to induce hepatotox-icity in humans, and 13 were reported to also cause steatosis. In order to compare in vivo relevant blood con-centrations, pharmacokinetic models were established for all compounds to simulate the maximal blood concentrations (Cmax) at therapeutic doses. The results showed that several hepatotoxic compounds induced an increase in lipid droplets at sub-cytotoxic concentrations. To compare how well (1) the cytotoxicity test alone, (2) the Nile Red assay alone, and (3) the combination of the cytotoxicity test and the Nile Red assay (based on the lower EC10 of both assays) allow the differentiation between hepatotoxic and non-hepatotoxic compounds, a previously established performance metric, the Toxicity Separation Index (TSI) was calculated. In addition, the Toxicity Estimation Index (TEI) was calculated to determine how well blood concentrations that cause an increased DILI risk can be estimated for hepatotoxic compounds. Our findings indicate that the combination of both assays improved the TSI and TEI compared to each assay alone. In conclusion, the study demonstrates that inclusion of the Nile Red assay into in vitro test batteries may improve the prediction of DILI compounds.

Datos de la publicación

ISSN/ISSNe:: 0378-4274, 1879-3169
Tipo:: Article
Páginas:: 33-46
DOI:: 10.1016/j.toxlet.2022.08.006
PubMed:: 35963427
Factor de Impacto:: 0,804 SCImago ℠
Cuartil:: Q1 SCImago ℠

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Filiaciones

Brecklinghaus, Tim:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Albrecht, Wiebke:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Duda, Julia:: Department of Statistics, TU Dortmund University, Vogelpothsweg 87, 44227, Dortmund, Germany
Kappenberg, Franziska:: Department of Statistics, TU Dortmund University, Vogelpothsweg 87, 44227, Dortmund, Germany
Gruendler, Lisa:: Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, Ardeystr 67, D-44139 Dortmund, Germany
Edlund, Karolina:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Marchan, Rosemarie:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Ghallab, Ahmed:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany

Department of Forensic Medicine and Toxicology, Faculty of Veterinary Medicine, South Valley University, 83523 Qena, Egypt
Cadenas, Cristina:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Rieck, Adrian:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Vartak, Nachiket:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany
Tolosa, Laia:: Instituto de Investigación. Experimental Hepatology Unit, Health Research Institute La Fe, Valencia, Spain
Gardner, Iain:: Simcyp (A Certara Company), Sheffield, UK
Halilbasic, Emina:: Hans Popper Laboratory of Molecular Hepatology, Division of Gastroenterology and Hepatology, Department of Internal Medicine III, Medical University of Vienna, Vienna, Austria
Trauner, Michael:: Hans Popper Laboratory of Molecular Hepatology, Division of Gastroenterology and Hepatology, Department of Internal Medicine III, Medical University of Vienna, Vienna, Austria
Ullrich, Anett:: Primacyt Cell Culture Technology GmbH, Schwerin, Germany
Zeigerer, Anja:: Institute for Diabetes and Cancer, Helmholtz Center Munich, 85764 Neuherberg, Germany

German Center for Diabetes Research (DZD), 85764 Neuherberg, Germany

Joint Heidelberg-IDC Translational Diabetes Program, Inner Medicine 1, Heidelberg University Hospital, Heidelberg, Germany
Turgunbayer, Oezlem Demirci:: Tech Univ Dortmund IfADo, Leibniz Res Ctr Working Environm & Human Factors, Ardeystr 67, D-44139 Dortmund, Germany

Dicle Univ, Fac Sci, Dept Biol, TR-21280 Diyarbakir, Turkey
Damm, Georg:: Department of Hepatobiliary Surgery and Visceral Transplantation, University of Leipzig, 04103 Leipzig, Germany
Seehofer, Daniel:: Department of Hepatobiliary Surgery and Visceral Transplantation, University of Leipzig, 04103 Leipzig, Germany
Rahnenfuhrer, Joerg:: Department of Statistics, TU Dortmund University, Vogelpothsweg 87, 44227, Dortmund, Germany
Hengstler, Jan G.:: Leibniz Research Centre for Working Environment and Human Factors at the Technical University of Dortmund (IfADo), Ardeystrasse 67, 44139 Dortmund, Germany

Keywords

Steatosis; DILI; Alternative methods; NAFLD; Lipid droplets

Proyectos y Estudios Clínicos

DESARROLLO DE UN MODELO HEPATOCELULAR HUMANO DIFERENCIADO PARA ESTUDIOS DE METABOLISMO Y POTENCIAL INDUCTOR DE NUEVOS FARMACOS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

SAF2003-09353 . 2003

A-CUTE-TOX

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

LSHB-CT-2004-512051-ACUTE TOX . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

DESARROLLO DE NUEVAS ESTRATEGIAS DIAGNOSTICAS BASADAS EN EL ESTUDIO DE LA CAPACIDAD METABOLIZANTE DE LA PIEL Y DEL PAPEL DE LAS CELULAS DENDITRICAS EN LA INDUCCION DE LAS REACCIONES ALERGICAS FRENTE A FARMACOS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

PARTIC PI05/2290 - FUND IMABIS . INSTITUTO DE SALUD CARLOS III; FUNDACION FIMABIS . 2007

DEVELOPMENT OF A HIGH THROUGHPUT GENOMICS-BASED TEST FOR ASSESSING GENOTOXIC AND CARCINOGENIC PROPERTIES OF CHEMICAL COMPOUNDS IN VITRO

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

CARCINOGENOMICS . COMISION EUROPEA . 2006

LINTOP

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

LSHB-CT-2006-037499 - LINTOP . FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2005

MECANISMOS TRNACRIPCIONALES IMPLICADOS EN LA ETIOLOGIA DEL HIGADO GRASO NO ALCOHOLICO. ESTUDIOS EN UN MODELO CELULAR HUMANO DE ESTEATOSIS Y APLICACION AL TRANSPLANTE DE HEPATOCITOS EN TERAPIA CELULAR.

Investigador Principal: RAMIRO JOVER ATIENZA

PI07/0550 . INSTITUTO DE SALUD CARLOS III . 2007

TU HOSPITAL INVESTIGA PARA TI

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

CCT005-07-00936 . 2007

PROYECTO CAIBER

CAI08/01/0061 . INSTITUTO DE SALUD CARLOS III; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2009

FORINVITOX

Investigador Principal: MARÍA JOSÉ GÓMEZ-LECHÓN MOLINER

FORINVITOX 1, START UP 2 . REMA; UNIVERSIDAD AUTONOMA BARCELONA; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2008

RED DE INNOVACION EN TECNOLOGIAS MEDICAS Y SANITARIAS - RED ITEMAS

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

RD09/0077/00156 . INSTITUTO DE SALUD CARLOS III . 2010

CREACION Y MEJORA DE INFRAESTRUCTURAS EN MEDICINA REGENERATIVA

Investigador Principal: FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA

PLE2009-0141 ACI-PLAN E . MINISTERIO DE CIENCIA E INNOVACION; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2009

MECANISMOS TRANSCRIPCIONALES IMPLICADOS EN EL HIGADO GRASO NO ALCOHOLICO DE ORIGEN METABOLICO E IATROGENICO: INFLUENCIA DE LA RESISTENCIA A LA INSULINA

Investigador Principal: RAMIRO JOVER ATIENZA

PI10/00194 . INSTITUTO DE SALUD CARLOS III . 2010

METABONOMICA Y CITOMICA COMO NUEVOS PARADIGMAS Y ESTRATEGIAS EXPERIMENTALES PARA EL ESTUDIO DE LOS MECANISMOS DE HEPATOTOXICIDAD DE FARMACOS, PREDICCION DE SU POTENCIAL HEPATOTOXICO Y LA TRANSICION DE HEPATITIS I ATROGENICA A AUTOINMUNE.

Investigador Principal: MARÍA TERESA DONATO MARTÍN

PI10/00923 . INSTITUTO DE SALUD CARLOS III . 2010

INNOVALIV: INNOVATIVE STRATEGIES TO GENERATE HUMAN HEPATOCYTES FOR TREATMENT OF METABOLIC LIVER DISEASES: TOOLS FOR PERSONALIZED CELL THERAPY

Investigador Principal: JOSÉ VICENTE CASTELL RIPOLL

278152_INNOVALIV_PE_CASTELL_FP7-HEALTH-2011-two-st . COMISION EUROPEA . 2011

PROGRAMA DE TECNICO DE APOYO. MODALIDAD EMPRESAS

Investigador Principal: FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA

PTA2012-8060-E . MINISTERIO DE ECONOMIA Y COMPETITIVIDAD . 2013

SOLICITUD DE CENTRO I+D RAMON Y CAJAL

Investigador Principal: MARÍA ÁNGELES TORMO MAS

RYC-2012-12246 . MINISTERIO DE ECONOMIA Y COMPETITIVIDAD; FUNDACIÓN PARA LA INVESTIGACIÓN DEL HOSPITAL UNIVERSITARIO LA FE DE LA COMUNIDAD VALENCIANA . 2014