Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications

Leonard J. Nelson, Katie Morgan, Philipp Treskes, Kay Samuel, Catherine J. Henderson, Claire LeBled, Natalie Homer, M. Helen Grant, Peter C. Hayes, John N. Plevris

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Conventional in vitro human hepatic models for drug testing are based on the use of standard cell lines derived from hepatomas or primary human hepatocytes (PHHs). Limited availability, inter-donor functional variability and early phenotypic alterations of PHHs restrict their use; whilst standard cell lines such as HepG2 lack a substantial and variable set of liver-specific functions such as CYP450 activity. Alternatives include the HepG2-derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human HepaRG cells are an alternative organotypic co-culture model of hepatocytes and cholangiocytes reported to maintain in vivo-like liver-specific functions, including intact Phase 1-3 drug metabolism. In this study, we compared C3A and human HepaRG cells using phenotypic profiling, CYP450 activity and drug metabolism parameters to assess their value as hepatic models for pre-clinical drug testing or therapeutics. Compared with C3As, HepaRG co-cultures, exhibit a more organotypic phenotype, including evidence of hepatic polarity with strong expression of CYP3A4, the major isoform involved in the metabolism of over 60% of marketed drugs. Significantly greater CYP450 activity and expression of CYP1A2, CYP2E1 and CYP3A4 genes in HepaRG cells (comparable with that of human liver tissue) was demonstrated. Moreover, HepaRG cells also preferentially expressed the hepatic integrin α5β1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin (CYP1A2) and testosterone (CYP3A4) using LC-MS/MS and HPLC, respectively, revealed HepaRGs had more intact (Phase 1-2) metabolism profile. Thus, HepaRG cells significantly outperform C3A cells for potential pharmaceutical and therapeutic applications.
LanguageEnglish
Pages30-37
Number of pages8
JournalBasic and Clinical Pharmacology and Toxicology
Volume120
Issue number1
Early online date10 Jun 2016
DOIs
Publication statusPublished - 31 Jan 2017

Fingerprint

Hep G2 Cells
Metabolism
Hepatocytes
Phenotype
Liver
Cytochrome P-450 CYP3A
Pharmaceutical Preparations
Cytochrome P-450 CYP1A2
Therapeutics
Coculture Techniques
Cells
Phenacetin
Cytochrome P-450 CYP2E1
Cell Line
Testing
Metabolites
Integrins
Modulators
Testosterone
Protein Isoforms

Keywords

  • human hepatic models
  • drug testing
  • primary human hepatocytes
  • human liver tissue
  • drug metabolite profiling

Cite this

Nelson, Leonard J. ; Morgan, Katie ; Treskes, Philipp ; Samuel, Kay ; Henderson, Catherine J. ; LeBled, Claire ; Homer, Natalie ; Grant, M. Helen ; Hayes, Peter C. ; Plevris, John N. / Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications. In: Basic and Clinical Pharmacology and Toxicology . 2017 ; Vol. 120, No. 1. pp. 30-37.
@article{c8484978b1cf4b2cad37c6800d2ac51f,
title = "Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications",
abstract = "Conventional in vitro human hepatic models for drug testing are based on the use of standard cell lines derived from hepatomas or primary human hepatocytes (PHHs). Limited availability, inter-donor functional variability and early phenotypic alterations of PHHs restrict their use; whilst standard cell lines such as HepG2 lack a substantial and variable set of liver-specific functions such as CYP450 activity. Alternatives include the HepG2-derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human HepaRG cells are an alternative organotypic co-culture model of hepatocytes and cholangiocytes reported to maintain in vivo-like liver-specific functions, including intact Phase 1-3 drug metabolism. In this study, we compared C3A and human HepaRG cells using phenotypic profiling, CYP450 activity and drug metabolism parameters to assess their value as hepatic models for pre-clinical drug testing or therapeutics. Compared with C3As, HepaRG co-cultures, exhibit a more organotypic phenotype, including evidence of hepatic polarity with strong expression of CYP3A4, the major isoform involved in the metabolism of over 60{\%} of marketed drugs. Significantly greater CYP450 activity and expression of CYP1A2, CYP2E1 and CYP3A4 genes in HepaRG cells (comparable with that of human liver tissue) was demonstrated. Moreover, HepaRG cells also preferentially expressed the hepatic integrin α5β1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin (CYP1A2) and testosterone (CYP3A4) using LC-MS/MS and HPLC, respectively, revealed HepaRGs had more intact (Phase 1-2) metabolism profile. Thus, HepaRG cells significantly outperform C3A cells for potential pharmaceutical and therapeutic applications.",
keywords = "human hepatic models, drug testing, primary human hepatocytes, human liver tissue, drug metabolite profiling",
author = "Nelson, {Leonard J.} and Katie Morgan and Philipp Treskes and Kay Samuel and Henderson, {Catherine J.} and Claire LeBled and Natalie Homer and Grant, {M. Helen} and Hayes, {Peter C.} and Plevris, {John N.}",
year = "2017",
month = "1",
day = "31",
doi = "10.1111/bcpt.12631",
language = "English",
volume = "120",
pages = "30--37",
journal = "Basic and Clinical Pharmacology and Toxicology",
issn = "1742-7835",
number = "1",

}

Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications. / Nelson, Leonard J.; Morgan, Katie; Treskes, Philipp; Samuel, Kay; Henderson, Catherine J.; LeBled, Claire; Homer, Natalie; Grant, M. Helen; Hayes, Peter C.; Plevris, John N.

In: Basic and Clinical Pharmacology and Toxicology , Vol. 120, No. 1, 31.01.2017, p. 30-37.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Human hepatic HepaRG cells maintain an organotypic phenotype with high intrinsic CYP450 activity/metabolism and significantly outperform standard HepG2/C3A cells for pharmaceutical and therapeutic applications

AU - Nelson, Leonard J.

AU - Morgan, Katie

AU - Treskes, Philipp

AU - Samuel, Kay

AU - Henderson, Catherine J.

AU - LeBled, Claire

AU - Homer, Natalie

AU - Grant, M. Helen

AU - Hayes, Peter C.

AU - Plevris, John N.

PY - 2017/1/31

Y1 - 2017/1/31

N2 - Conventional in vitro human hepatic models for drug testing are based on the use of standard cell lines derived from hepatomas or primary human hepatocytes (PHHs). Limited availability, inter-donor functional variability and early phenotypic alterations of PHHs restrict their use; whilst standard cell lines such as HepG2 lack a substantial and variable set of liver-specific functions such as CYP450 activity. Alternatives include the HepG2-derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human HepaRG cells are an alternative organotypic co-culture model of hepatocytes and cholangiocytes reported to maintain in vivo-like liver-specific functions, including intact Phase 1-3 drug metabolism. In this study, we compared C3A and human HepaRG cells using phenotypic profiling, CYP450 activity and drug metabolism parameters to assess their value as hepatic models for pre-clinical drug testing or therapeutics. Compared with C3As, HepaRG co-cultures, exhibit a more organotypic phenotype, including evidence of hepatic polarity with strong expression of CYP3A4, the major isoform involved in the metabolism of over 60% of marketed drugs. Significantly greater CYP450 activity and expression of CYP1A2, CYP2E1 and CYP3A4 genes in HepaRG cells (comparable with that of human liver tissue) was demonstrated. Moreover, HepaRG cells also preferentially expressed the hepatic integrin α5β1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin (CYP1A2) and testosterone (CYP3A4) using LC-MS/MS and HPLC, respectively, revealed HepaRGs had more intact (Phase 1-2) metabolism profile. Thus, HepaRG cells significantly outperform C3A cells for potential pharmaceutical and therapeutic applications.

AB - Conventional in vitro human hepatic models for drug testing are based on the use of standard cell lines derived from hepatomas or primary human hepatocytes (PHHs). Limited availability, inter-donor functional variability and early phenotypic alterations of PHHs restrict their use; whilst standard cell lines such as HepG2 lack a substantial and variable set of liver-specific functions such as CYP450 activity. Alternatives include the HepG2-derivative C3A cells selected as a more differentiated and metabolically active hepatic phenotype. Human HepaRG cells are an alternative organotypic co-culture model of hepatocytes and cholangiocytes reported to maintain in vivo-like liver-specific functions, including intact Phase 1-3 drug metabolism. In this study, we compared C3A and human HepaRG cells using phenotypic profiling, CYP450 activity and drug metabolism parameters to assess their value as hepatic models for pre-clinical drug testing or therapeutics. Compared with C3As, HepaRG co-cultures, exhibit a more organotypic phenotype, including evidence of hepatic polarity with strong expression of CYP3A4, the major isoform involved in the metabolism of over 60% of marketed drugs. Significantly greater CYP450 activity and expression of CYP1A2, CYP2E1 and CYP3A4 genes in HepaRG cells (comparable with that of human liver tissue) was demonstrated. Moreover, HepaRG cells also preferentially expressed the hepatic integrin α5β1 – an important modulator of cell behaviour including growth and survival, differentiation and polarity. Drug metabolite profiling of phenacetin (CYP1A2) and testosterone (CYP3A4) using LC-MS/MS and HPLC, respectively, revealed HepaRGs had more intact (Phase 1-2) metabolism profile. Thus, HepaRG cells significantly outperform C3A cells for potential pharmaceutical and therapeutic applications.

KW - human hepatic models

KW - drug testing

KW - primary human hepatocytes

KW - human liver tissue

KW - drug metabolite profiling

U2 - 10.1111/bcpt.12631

DO - 10.1111/bcpt.12631

M3 - Article

VL - 120

SP - 30

EP - 37

JO - Basic and Clinical Pharmacology and Toxicology

T2 - Basic and Clinical Pharmacology and Toxicology

JF - Basic and Clinical Pharmacology and Toxicology

SN - 1742-7835

IS - 1

ER -