The Pharma-BAL: A human liver model for in vitro studies

Ronde 2012 Module Proefdiervrije Technieken: Er wordt veel onderzoek gedaan naar leverinfecties en behandelmethodes daarvan en naar de veiligheid van medicijnen op de lever. De levercellijnen die hiervoor meestal gebruikt worden hebben een te lage functionaliteit en daarom worden ook proefdieren gebruikt. Dit MKMD-project had als doel de levercelkweek te verbeteren. We hebben het DNA dat codeert voor een regulator van ontgifting en energiehuishouding, CAR, ingebracht in de menselijke levercellijn HepaRG. De ontstane nieuwe cellijn, HepaRG-CAR, vertoonde verbeterde ontgifting en energiehuishouding, maar ook onverwachte eigenschappen, zoals hogere bloedeiwit productie. In combinatie met een nieuw ontwikkeld kweeksysteem, waarbij de cellen in 3D en onder mediumperfusie worden gekweekt, werd nog een hogere functionaliteit bereikt. Deze combinatie kan in de toekomst ingezet worden voor meer betrouwbaar leveronderzoek en, als consequentie, het gebruik van proefdieren terugdringen.
Titel: A perfused oxygenated bio-artificial liver containing HepaRG cells in a three-dimensional configuration as an improved model for drug hepatotoxicity and metabolism studies.
Auteur: Vincent A. van der Mark, Martien van Wenum, D. Rudi de Waart, David Steen, Valery Shevchenko, Robert A.F.M. Chamuleau, Christophe Chesné, Ruurdtje Hoekstra
Titel: PharmaBAL: an advanced model for drug metabolism and hepatotoxicity studies
Auteur: Van der Mark, V.
Titel: Overexpression of the constitutive androstane receptor and culture in a novel 3D/perfusion system increases hepatic differentiation and oxidative metabolism in HepaRG cells
Auteur: Vincent van der Mark, Rudi de Waart, Valery Shevchenko, Ronald Oude Elferink, Robert Chamuleau, Ruurdtje Hoekstra
Titel: Stable Overexpression of the Constitutive Androstane Receptor Reduces the Requirement for Culture with Dimethyl Sulfoxide for High Drug Metabolism in HepaRG Cells
Auteur: van der Mark, V. A., Rudi de Waart, D., Shevchenko, V., Elferink, R. P. J. O., Chamuleau, R. A. F. M., Hoekstra, R.
Magazine: Drug Metabolism and Disposition
Titel: Hepatic cell line resistant to dimethyl sulfoxide, cell culture and uses thereof.
Auteur: Van der Mark, V.A. and Hoekstra, R.

Samenvatting van de aanvraag
The liver is a central organ that is essential for the control of body homeostasis, detoxification and synthesis of blood proteins. As the target organ for many diseases and adverse effects of drugs, the liver is one of the most studied organs. A towering number of animals are sacrificed for these studies, including studies on hepatitis B virus (HBV) and liver toxicity, although the predictability of many animal models is limited. Thus the need for alternative methods that mimic the human liver is high. A well defined bioartificial liver (BAL) might be a worthwhile substitute for many animal experiments. An AMC program on BALs has resulted in the development of a bioreactor that is optimised for the human liver cell line HepaRG. The interaction between the bioreactor and the cell line yields an all-round high hepatic functionality, reproducibility and stability, outperforming all other existing in vitro liver models. The ZonMw MKMD grant will be applied to finance a public-private partnership dedicated to transform the model into a marketable and practical Pharma-BAL kit, ready for application as a HBV culture model and for implementation in drug safety study programs of the pharmaceutical industry. To reach this goal a strong public-private partnership will be created from complementary partners: two academic groups (from AMC and Heidelberg University) and two SMEs (AMC spin-off company Hep-Art Medical Devices and Biopredic, Rennes). Together they bring in their expertise and IP on bioreactors (AMC, Hep-Art), the cell line HepaRG (Biopredic, AMC), pharmacological testing (Biopredic) and HBV (Heidelberg), in addition to experience in business development (Biopredic, Hep-Art) and an established network for commercialization and distribution of cells for compound safety studies (Biopredic). The 3-year project is organized into 5 work packages: 1. Miniaturization: Downscaling of the current bioreactor from 9 mL to 2 mL 2. Product description: Testing relevant parameters for application as a model for HBV infections and drug toxicity: a. HBV infectivity experiments including a pilot phase, and if successful, followed by a characterization of the HBV life cycle in the Pharma-BAL and finally showing proof of principle by testing the effects of drugs targeted to different phases of HBV infectivity. b. Drug toxicity studies including a characterization of the drug metabolism followed by a characterization of the cytotoxic effects of drugs known to elicit specific toxic effects, as cholestasis, steatosis etc. 3. Logistics: Establishing optimal preservation and bioreactor loading conditions 4. Multiplexing: Developing a set-up for managing up to 10 Pharma-BALs simultaneously 5. Business development: Development of the Pharma-BAL into a commercial product The consortium is able to deliver a human liver model to be used by industries and research organizations for their investigations of HBV infections and therapies targeted against this virus, and by pharmaceutical industries in the safety assessment of new developed-drugs. For this project a € 300.000 ZonMw contribution is required and the project group will match in kind for another € 75.000. This budget is sufficient to yield a marketable product as the technical feasibility of the project is high, the project group members are experienced and highly qualified, the route to the market is relatively short and end-users will be actively involved in the project. Therefore the chance of successful outcome of the project is high. As a result, implementation of the Pharma-BAL technology will reduce considerably the number of animal experiments for studying drug safety and human liver diseases.
Onderdeel van programma:
Gerelateerde subsidieronde:
Projectleider en penvoerder:
Dr. R. Hoekstra
Verantwoordelijke organisatie:
Amsterdam UMC - locatie AMC