Integrated Approach to Cyclopiazonic Acid Cytotoxicity Using In Vitro (2D and 3D Models) and In Silico Methods

Cyclopiazonic acid (CPA) is an indole-tetramic acid neurotoxin produced by <i>Aspergillus</i> and <i>Penicillium</i> genera present mainly in fruit, cereals and nuts. This study compares the cytotoxicity produced by CPA after 24, 48 and 72 h of exposure using both monolayers and 3D spheroids in human neuroblastoma SH-SY5Y cells. Furthermore, CPA toxicokinetics was evaluated using <i>in silico</i> models. Cytotoxicity increased dose- and time-dependently, as shown by the MTT assay. The lowest CPA IC₅₀ values were found in the monolayer study compared to the 3D spheroids at all exposure times (24 h: 864.01 vs. 1132; 48 h: 437 vs. 1069; 72 h: 392 vs. 567 nM). The CPA exposure on SH-SY5Y spheroid organization and morphology was also studied. Morphological changes, including spheroid disaggregation, were observed after mycotoxin exposure. The <i>in silico</i> methods, SwissADME and admetSAR, were used for short and full ADMEt profiles of CPA. The ADMEt predictive profile shows high gastrointestinal absorption and ability to penetrate the blood-brain barrier. Including <i>in silico</i> studies emphasizes the comprehensive approach to understanding mycotoxin toxicity and risk assessment. By combining <i>in vitro</i> 3D spheroid models with computational simulations, this study aims to provide a holistic perspective on the effects of CPA, enhancing the accuracy and relevance of our findings.