Establishment of in vitro cholestatic drug-induced liver injury evaluation system using HepG2-hNTCP-hBSEP-C4 cells.

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K16051
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47060:Clinical pharmacy-related
• Research Institution: Nagoya City University
• Principal Investigator: Sakai Yoko 名古屋市立大学, 大学院薬学研究科, 助教 (50723079)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Keywords: 毛細胆管 / bile salt export pump / 凍結ヒト肝細胞 / 薬剤性胆汁うっ滞肝障害 / カスパーゼ / MPS

Outline of Final Research Achievements

Although laboratory animals are used, the use of human-derived cells is required due to species differences. One of the problems with primary human hepatocytes (HPHs) is the rapid loss of function after culture, and difficult to form bile canalicular. We investigated a culture method for efficient formation of bile canalicular in primary human hepatocytes (HPHs). The results indicated that HPHs cultured with Z-VAD-FMK, a total caspase inhibitor, and RevitaCellTM Supplement, an irreversible ROCK inhibitor, in combination with RM-101 from 1 hr after thawing to day 2 formed bile canalicular on day 6 of culture. The expression on F-actin in bile salt export pump (BSEP) was observed and it also had a function. In additionally, HPHs with a novel method cultured on the device were able to predict cholestatic DILI risk.

Free Research Field

医療薬学

Academic Significance and Societal Importance of the Research Achievements

薬剤性胆汁鬱滞型肝障害評価系の構築のために、主な胆汁酸排泄トランスポーターであるBSEPの発現や機能にも焦点を当てて研究しており、BSEPの機能不全が起こる原因やその疾患にも応用することができる。また、本研究室で開発したデバイスと組み合わせて、培地を灌流した薬剤性胆汁鬱滞型肝障害評価系を開発することにも新規性がある。医薬品候補化合物の毒性を見極めるスクリーニングにおいて安価かつ容易に大量生産可能であり、短期間に評価可能であるため、安全性の高い医薬品の開発の礎になると思われる。