5-ALA treatment increases intracellular heme levels and enhances CYP3A4 activity in genome-edited Caco-2 cells

• Published in: Biochemical and biophysical research communications Vol. 664; pp. 94 - 99
• Main Authors: Watanabe, Keita, Negoro, Ryosuke, Fujita, Takuya
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 05.07.2023
• Subjects: 5-ALA; Aminolevulinic Acid - pharmacology; Caco-2 Cells; Caco-2 cell; CYP3A4; Cytochrome P-450 CYP3A - genetics; Cytochrome P-450 CYP3A - metabolism; Cytochrome P-450 Enzyme System - metabolism; Drug-metabolizing enzyme; Heme; Humans; Drug-metabolizing enzyme; Caco-2 cell; CYP3A4; 5-ALA
• ISSN: 0006-291X; 1090-2104
• DOI: 10.1016/j.bbrc.2023.04.077

In nonclinical studies, models that can predict the metabolism of drug candidates by cytochrome P450 (CYP), including Cytochrome P450 family 3 subfamily A member 4 (CYP3A4) are helpful. CYP3A4-overexpressing human cells have been used universally to evaluate whether CYP3A4 metabolizes drug-candidate compounds. However, CYP3A4-overexpressing human cell lines are problematic because their activity levels are lower than that of in vivo human CYP3A4. Heme plays a paramount role in CYP activity. The rate-limiting step in heme biosynthesis is the generation of 5-aminolevulinic acid (5-ALA). In this study, we examined whether treatment with 5-ALA to CYP3A4-POR-UGT1A1-CES2 knockin and CES1 knockout (genome-edited) Caco-2 cells enhances CYP3A4 activity. A 7-day 5-ALA treatment increased intracellular heme levels in genome-edited Caco-2 cells without cytotoxicity. Moreover, consistent with the increase in intracellular heme content, 5-ALA treatment increased CYP3A4 activity in genome-edited Caco-2 cells. The results of this research are expected to be applied to pharmacokinetic studies using CYP-overexpressing human cells containing CYP3A4. •5-ALA treatment increased intracellular heme levels in genome-edited Caco-2 cells.•Long-term 5-ALA treatment in genome-edited Caco-2 cells resulted in toxicity.•5-ALA treatment increased CYP3A4 activity in genome-edited Caco-2 cells.