Comprehensive DNA Methylation Analysis of Human Neuroblastoma Cells Treated With Haloperidol and Risperidone

Abstract

Accumulating evidence suggests that the epigenetic alterations induced by antipsychotics contribute to the therapeutic efficacy. However, global and site-specific epigenetic changes by antipsychotics and those shared by different classes of antipsychotics remain poorly understood. We conducted a comprehensive DNA methylation analysis of human neuroblastoma cells cultured with antipsychotics. The cells were cultured with low and high concentrations of haloperidol or risperidone for 8 days. DNA methylation assay was performed with the Illumina HumanMethylation450 BeadChip. We found that both haloperidol and risperidone tended to cause hypermethylation changes and showed similar DNA methylation changes closely related to neuronal functions. A total of 294 differentially methylated probes (DMPs), including 197 hypermethylated and 97 hypomethylated DMPs, were identified with both haloperidol and risperidone treatment. Gene ontology analysis of the hypermethylated probe-associated genes showed enrichment of genes related to the regulation of neurotransmitter receptor activity and lipoprotein lipase activity. Pathway analysis identified that among the DMP-associated genes, SHANK1 and SHANK2 were the major genes in the neuropsychiatric disorder-related pathways. Our data would be valuable for understanding the mechanisms of action of antipsychotics from an epigenetic viewpoint.

Assessment of Protective Effects of Carvacrol on Haloperidol-Induced Oxidative Stress and Genotoxicity in Human Peripheral Blood Lymphocytes

“…Based on the results of this study, exposing human lymphocytes to different concentrations of haloperidol (25, 50, and 100 ng/ml) resulted in significant increases in the frequency of MN, DNA percentage in tail, and tail moment, indicating that haloperidol might have the potential to induce genotoxicity. This is consistent with the results reported in previous studies [ 36 , 47 – 51 ]. In a similar study, the CBMN test and alkaline comet assay were conducted on human lymphocytes at lower concentrations of haloperidol (5, 10 and 20 ng/ml) compared to our study.…”

“…The results of a recent study also revealed that haloperidol may potentiate the induction of DNA methylation in neuroblastoma cells. It was hypothesized that inducing DNA methylation is associated with haloperidol's strong blockade of dopamine D2 receptors [ 51 ]. The blockade of dopaminergic transmission by haloperidol leads to an increase in the turnover rate of dopamine, which will consequently result in the excessive production of reactive oxygen species and oxidative stress [ 52 ].…”

Neurotoxic/Neuroprotective Effects of Clozapine and the Positive Allosteric Modulator of mGluR2 JNJ-46356479 in Human Neuroblastoma Cell Cultures

“…Interestingly, some mGluR2/3 agonists have previously shown neuroprotection against different neurotoxic insults in different cell cultures [22], including neuroblastoma [23,24]. The human neuroblastoma cell line displays neuronal properties and is a popular in vitro model used in neuropsychiatric research including schizophrenia [23][24][25][26] that has been widely used to test pharmacological effects of drugs including APs [26][27][28][29].…”

Antipsychotics function as epigenetic age regulators in human neuroblastoma cells

“…We previously performed comprehensive DNA methylation assays of human neuroblastoma cells cultured with various antipsychotics and revealed that the antipsychotics induced similar epigenetic signatures 22 – 24 . Although experiments using cell lines had several limitations in that they acquired immortal, cancer-like characteristics and did not reflect complex synaptic networks in the brain, they provided the ideal opportunity to test the effects of antipsychotics in vitro.…”

“…For each antipsychotic, two doses (high and low), which were determined based on their effective blood concentrations, were tested with each DNA methylation assay. The assays using HAL and RIS 22 and those using BL and PE 23 , 24 were conducted over different periods of time. Although we used the same source of cell lines and employed the same experimental protocol, principal component analysis suggested batch effect differences between datasets (Fig.…”