Tracking Transcriptomic Dynamics in HepG2/C3A Spheroids: Implications for Drug Toxicity Research

Abstract

The study of transcriptomic dynamics in three-dimensional (3D) hepatic spheroids has gained significant attention due to their relevance in drug toxicity screening. This research investigates the transcriptomic changes in HepG2/C3A spheroids over time, offering insights into their potential as an in vitro model for hepatotoxicity assessment. HepG2/C3A cells, a subclone of HepG2, possess improved liver-like functionalities, making them a valuable model for drug metabolism studies. By employing RNA sequencing (RNA-seq) at multiple time points, we analyzed gene expression patterns to capture the cellular response to prolonged culture in a 3D microenvironment. We observed differential gene expression associated with metabolic pathways, stress responses, and xenobiotic metabolism, indicating a dynamic adaptation of the spheroids to their environment. Additionally, exposure to known hepatotoxic compounds revealed transcriptomic signatures indicative of early toxicity responses. These findings underscore the relevance of HepG2/C3A spheroids in toxicological evaluations and highlight the importance of tracking their transcriptomic evolution over time. The study provides a foundation for refining in vitro liver models, enhancing drug safety assessments, and reducing reliance on animal testing.