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Establishment of MDR1-knockout human enteroids for pharmaceutical application
Tatsuya Inui1, Ryuga Nomoto1, Jumpei Yokota1
1Laboratory of Biochemistry and Molecular Biology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, 565-0871, Japan.
Drug Metabolism and Pharmacokinetics
|December 15, 2022
Summary
This study introduces a novel human enteroid model using genome editing to overcome limitations in drug transporter research. This advanced model accurately assesses drug transporter contributions, improving drug development safety and efficacy.
Area of Science:
- Pharmacokinetics and Drug Metabolism
- Genomics and Molecular Biology
- Gastroenterology and Drug Delivery
Background:
- Assessing drug transporters and metabolizing enzymes is crucial for intestinal pharmacokinetics in drug development.
- Conventional in vitro systems like Caco-2 cells exhibit low gene expression and inhibitors have off-target effects.
- Existing methods struggle to accurately evaluate the specific roles of pharmacokinetic-related genes.
Purpose of the Study:
- To establish a more accurate in vitro model for evaluating drug transporter contributions to intestinal pharmacokinetics.
- To overcome the limitations of low gene expression and off-target inhibitor effects in current models.
- To utilize genome editing technology in human enteroids for precise assessment of drug transporter functions.
Main Methods:
- Established human biopsy-derived enteroids with deficiency in MDR1 (multidrug resistance protein 1), a key efflux transporter.
- Utilized genome editing to create MDR1-knockout (KO) enteroids.
- Cultured enteroids into monolayers and assessed expression/activity of pharmacokinetic genes (e.g., CYP3A4) and transporter functions.
Main Results:
- MDR1-KO enteroid-derived monolayers maintained high expression and activity of other pharmacokinetic genes like CYP3A4.
- Accurately evaluated MDR1's contribution to vinblastine cytotoxicity, a substrate metabolized by CYP3A4.
- Demonstrated that conventional methods using verapamil (an MDR1 inhibitor) failed due to its off-target inhibition of CYP3A4.
Conclusions:
- Human enteroid-derived monolayers combined with genome editing offer a powerful tool for evaluating specific pharmacokinetic molecules.
- This model precisely assesses the contribution of efflux transporters like MDR1, overcoming limitations of chemical inhibitors.
- The developed model enhances the accuracy of in vitro drug interaction and pharmacokinetic assessments in drug development.
