Small Molecule Inhibition of Protein Disulfide Isomerase in Neuroblastoma Cells Induces an Oxidative Stress Response and Apoptosis Pathways.

Protein disulfide isomerase (PDI) is a multifunctional enzyme located in the endoplasmic reticulum (ER) contributing to redox homeostasis and oxidative protein folding. PDI is associated with many diseases including neurodegenerative disorders like Alzheimer's disease and, hence, is considered a promising drug target. In this study, we investigate the abscisic acid (ABA)-derived PDI inhibitor origamicin for its neuropharmacological potential. First, we validated the function of origamicin by monitoring the inhibition of PDI's oxidoreductase activity using an in vitro enzyme activity assay. We also applied Huisgen cycloaddition chemistry (or "click chemistry") to interrogate the interaction of origamicin and PDI. Then, we evaluated the impact of origamicin on the viability of the neuroblastoma cell line SH-SY5Y. Next, we analyzed the gene expression profile of SH-SY5Y cells upon treatment with origamicin. We found 207 differentially expressed genes, including MYC. Computational analysis revealed an enrichment of genes involved in the oxidative stress response and the p53 signaling pathway. Induction of the p53 signaling pathway and downregulation of MYC are known to affect processes such as cell cycle, cellular repair, and apoptosis. Our study reveals the molecular mechanism of PDI inhibition by origamicin. Furthermore, this study provides important gene expression profiles that offer insights into the underlying mechanism of PDI inhibition and creates a valuable starting point for neuropharmacological applications of origamicin and other PDI inhibitors.