Sertindole, a Potent Antagonist at Dopamine D₂ Receptors, Induces Autophagy by Increasing Reactive Oxygen Species in SH-SY5Y Neuroblastoma Cells

Abstract

Autophagy is associated with cell survival and cell death. Autophagy is implicated in the pathophysiology of various human diseases. In order to identify autophagy regulatory molecules, we screened a chemical drug library in SH-SY5Y cells and selected Sertindole as a potent autophagy inducer. Sertindole was developed as an antipsychotic drug for Schizophrenia. Sertindole treatment highly induced the formation of autophagosomes as well as LC3 conversion. Subsequently, Sertindole-induced autophagy was efficiently suppressed by down regulation of ATG5. Sertindole also increased reactive oxygen species (ROS) production, which contributes to autophagy-associated cell death in neuroblastoma cells. ROS scavengers such as Nacetylcysteine and Trolox suppressed not only ROS generation but also autophagy activation by Sertindole. These results suggest Sertindole induces autophagy and autophagy-associated cell death by ROS production in neuroblastoma cells.Key words Sertindole; autophagy; ATG5; reactive oxygen species; SH-SY5Y cell Autophagy refers to a highly conserved lysosome-dependent degradation mechanism that involves the removal of cytoplasmic proteins and organelles.1,2) Autophagy plays important role in development, immune defense, and cell death. The fundamental role of autophagy helps cell survival under stressful conditions by enhancing metabolic homeostasis. However, autophagy may also contribute to non-apoptotic cell death processes under some conditions. 3-6) Indeed, autophagy has been implicated in many pathophysiologic conditions such as neurodegenerative diseases such as cancer, type 2 diabetes, infectious diseases, cardiomyopathies, and innate immune diseases.3-6) Therefore, understanding the molecular mechanism of autophagy may help to develop new therapeutic strategies for those diseases. Based on this notion, several groups reported on small molecules that modulate autophagy and can be used as potential therapeutic drugs. 7,8) For example, Rapamycin, an autophagy inducer, enhances clearance of intracellular aggregate proteins associated with Huntington's disease. 9,10) In addition, some autophagy regulators sensitize tumor cells to anticancer therapy. 11) However, the precise mechanisms of autophagy in pathological conditions are not fully understood.Schizophrenia (SZ) is a complex mental disorder often associated with cognitive impairment and depressive symptoms that affects about 1% of the population. SZ is caused by environmental and genetic risk factors or their interactions.12) Although, several genome wide association studies have identified SZ-related genes, the molecular mechanism is still largely unknown. [13][14][15] The primary treatment for SZ is antipsychotic medications.16,17) Sertindole, a phenylindole derivative was developed as an antipsychotic drug. It is a potent antagonist at dopamine D 2 , serotonin 5-HT 2 , and α 1 -adrenergic receptors with high affinity. 18,19) Additionally, Sertindole suppresses potassium channel in the brain and heart. 20,21) In contrast to other antipsychotic...

Clozapine Induces Autophagic Cell Death in Non-Small Cell Lung Cancer Cells

“…Recently, an autophagy assay system in SH-SY5Y neuroblastoma cells was built to screen 1120 chemicals and found that first-generation antipsychotics (pimozide, triflupromazine, chlorpromazine, and fluphenazine) and second-generation antipsychotics (such as sertindole) could induce autophagy. It was also discovered that sertindole can cause autophagic cell death in SH-SY5Y neuroblastoma cells [32], which is consistent with our findings in NSCLC cells.…”

Longitudinal Changes in Total Brain Volume in Schizophrenia: Relation to Symptom Severity, Cognition and Antipsychotic Medication

“…However, the mechanism through which this might occur is unclear, and it is conceivable that the differences between individual drugs may reflect the degree to which they may exert such effects [37]. In cell culture studies, a number of antipsychotics have been shown to induce autophagy, a process involved in neurodegenerative processes and cell death [38], suggesting a possible mechanism by which antipsychotics might cause loss of brain volume.…”

mTOR-Related Brain Dysfunctions in Neuropsychiatric Disorders

“…In contrast, chlorpromazine, which is a typical antipsychotic agent, induces autophagy by inhibiting the Akt/mTOR pathway [59]. Recently, in vitro studies on the effects of second-generation, atypical antipsychotics demonstrated that sertindole and clozapine are potent autophagy inducers in both neuronal and non-neuronal cell lines [257,261]. Similar to pimozide, clozapine activates the autophagy process via the AMPK–ULK1–Beclin1 pathway, as evidenced by increased levels of autophagy markers (i.e., LC3-II and Atg5–Atg12 conjugate); increased phosphorylation of AMPK and its downstream substrates, namely ULK1 and beclin1; and an increased number of autophagosomes in the frontal cortex in clozapine-treated rats [259].…”