Generation of predictive intestinal models of drug absorption, metabolism and toxicity using genome-editing technology.

2023 Fiscal Year Final Research Report

• Project/Area Number: 21K15264
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 47030:Pharmaceutical hygiene and biochemistry-related
• Research Institution: Ritsumeikan University
• Principal Investigator: Negoro Ryosuke 立命館大学, 薬学部, 助教 (10844045)
• Project Period (FY): 2021-04-01 – 2024-03-31
• Keywords: ゲノム編集 / CRISPR-Cas9 / Caco-2 cell / CYP3A4 / UGT1A1

Outline of Final Research Achievements

Caco-2 cells hardly express CYP3A4 and UGT1A1, drug-metabolizing enzymes that play a central role in drug metabolism, making it impossible to assess drug metabolism. This study attempted to solve the above problem by using genome-editing technology to induce high expression of CYP3A4 and UGT1A1 in Caco-2 cells. CYP3A4 and UGT1A1 activities in the genome-edited Caco-2 cells were extremely high compared to wild-type Caco-2 cells. These results suggest that I successfully generated stable CYP3A4 and UGT1A1-expressing Caco-2 cells using genome-editing technology.

Free Research Field

分子生物学

Academic Significance and Societal Importance of the Research Achievements

従来のCaco-2細胞では、薬物吸収はある程度予測できるものの、薬物代謝、薬物代謝に起因する毒性を予測することは困難であった。本研究で開発したCYP3A4、UGT1A1を高発現したCaco-2細胞は、従来のCaco-2細胞よりも優れた安全性評価モデルになりうる可能性を示した意義は大きい。また、CYP3A4、UGT1A1を高発現したCaco-2細胞は、培養コストが安価、継代・凍結保存が可能なため、創薬研究に要するコストカットにも貢献できる可能性がある。さらに、安全性試験における実験動物の使用削減につながるだろう。