AI-Guided Prediction of Aflatoxin B1 Bioactivation Validated by Toxicity and DNA Adduct Formation in VERO E6 Cells

Published

August 4, 2026

Author(s)

Bharti Sangwan, Ugochukwu Okoro, Isabella Atteck, Pawel Jaruga, Chinwe Ekenna, Michael Fasullo

Abstract

VERO cells, derived from the kidney epithelium of the African green monkey, are widely used in virology, but their ability to metabolize xenobiotics is not fully understood. Since cytochrome P450 (CYP) enzymes play a major role in both detoxification and activation of harmful compounds, we wanted to see whether these enzymes are functional in VERO-E6 cells. We started by looking at CYP gene expressions. Our results with Reverse Transcription- quantitative Polymerase Chain Reaction (RT-qPCR) showed that VERO-E6 cells express CYP3A4, CYP3A5, and CYP3A7. In contrast, CYP1A1, CYP2E1, and CYP2D6 were either very low or not detected under our conditions. To get an idea of whether these enzymes could activate aflatoxin B1 (AFB1), we used artificial intelligence (AI) -based structural modeling along with molecular docking. Based on these results, CYP3A enzymes appear to position AFB1 in a way that may support the formation of the reactive intermediate, and CYP3A4 showed the most favorable orientation. However, these predictions alone do not confirm whether DNA adducts are formed. To test this, we exposed VERO-E6 cells to 200 nmol/L AFB1. After 10 days, we observed about 40 % cell death. More importantly, liquid chromatography-tandem mass spectroscopy (LC–MS/MS) analysis confirmed the presence of AFB1-derived DNA adducts, indicating that metabolic activation is taking place in these cells. Taken together, these results suggest that VERO-E6 cells have functional CYP-mediated metabolic activity. From these results, it seems that combining computational and experimental approaches can help when studying xenobiotic metabolism, especially in systems like VERO cells, where detailed biochemical data are still limited.

Citation

Toxins

Volume

18

Issue

8

Pub Type

Journals

Download Paper

https://doi.org/10.3390/toxins18080339

Local Download

Keywords

Aflatoxin B1, AI-guided prediction, DNA adduct formation, VERO E6 cells

Citation

Sangwan, B. , Okoro, U. , Atteck, I. , Jaruga, P. , Ekenna, C. and Fasullo, M. (2026), AI-Guided Prediction of Aflatoxin B1 Bioactivation Validated by Toxicity and DNA Adduct Formation in VERO E6 Cells, Toxins, [online], https://doi.org/10.3390/toxins18080339, https://tsapps.nist.gov/publication/get_pdf.cfm?pub_id=962146 (Accessed September 24, 2026)

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