trichostatin-a and Schizophrenia

The novel technology of induced neuronal cells (iN cells) is promising for translational neuroscience, as it allows the conversion of human fibroblasts into cells with postmitotic neuronal traits. However, a major technical barrier is the low conversion rate. To overcome this problem, we optimized the conversion media. Using our improved formulation, we studied how major mental illness-associated chromosomal abnormalities may impact the characteristics of iN cells. We demonstrated that our new iN cell culture protocol enabled us to obtain more precise measurement of neuronal cellular phenotypes than previous iN cell methods. Thus, this iN cell culture provides a platform to efficiently obtain possible cellular phenotypes caused by genetic differences, which can be more thoroughly studied in research using other human cell models such as induced pluripotent stem cells.

Topics: Adolescent; Adult; Azacitidine; Cell Culture Techniques; Cell Differentiation; Chromosome Aberrations; Culture Media; Female; Fibroblasts; Humans; Hydroxamic Acids; Induced Pluripotent Stem Cells; Male; Middle Aged; Neural Stem Cells; Schizophrenia; Valproic Acid; Young Adult

The emerging field of psychiatric epigenetics is constrained by the dearth of research methods feasible in living patients. With this focus, we report on two separate approaches, one in vitro and one in vivo, developed in our laboratory.. In the first approach, we isolated lymphocytes from 12 subjects and cultured their cells with either 0.7 mM valproic acid (VPA), 100 nM Trichostatin A (TSA), or DMSO (control) for 24h based upon previous dose response experiments. We then measured GAD67 mRNA expression using realtime RT-PCR, total acetylated histone 3 (H3K9,K14ac) levels using Western blot analysis, and attachment of H3K9,K14ac to the GAD67 promoter using ChIP. In the second approach, we measured GAD67 mRNA and total H3K9,K14ac levels in lymphocytes from 11 schizophrenia and 7 bipolar patients before and after 4 weeks of clinical treatment with Depakote ER (VPA).. In the first approach, VPA induced a 383% increase in GAD67 mRNA, an 89% increase in total H3K9,K14ac levels, and a 482% increase in H3K9,K14ac attachment to the GAD67 promoter. TSA induced comparable changes on all measures. In the second approach, bipolar subjects had significantly higher baseline levels of H3K9,K14ac compared to subjects with schizophrenia. Subjects with clinically relevant serum levels of VPA (> or = 65 microg/mL) showed a significant increase in GAD67 mRNA expression.. Our results utilizing two separate approaches for examining chromatin remodeling in real clinical time provide possible means to investigate epigenetic events in living patients.

Topics: Acetylation; Adult; Bipolar Disorder; Blotting, Western; Cell Culture Techniques; Enzyme Inhibitors; Female; Gene Expression Regulation; Glutamate Decarboxylase; Histone Deacetylase Inhibitors; Histones; Humans; Hydroxamic Acids; Lymphocytes; Male; Middle Aged; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Schizophrenia; Up-Regulation; Valproic Acid; Young Adult

A restrictive chromatin state has been thought to be operant in the pathophysiology of schizophrenia. Our objective was to ascertain whether differences exist between baseline levels of a repressive chromatin mark such as dimethylated lysine 9 of histone 3 (H3K9me2) in patients with schizophrenia and healthy controls and whether a histone deacetylase (HDAC) inhibitor in an in vitro assay would differentially affect chromatin structure based on diagnosis.. We obtained blood samples from 19 healthy controls and 25 patients with schizophrenia and isolated their lymphocytes. We measured baseline H3K9me2 levels (normalized to total histone 1) in the lymphocytes from all participants via Western blot analysis. To examine the effects of an HDAC inhibitor on H3K9me2, we cultured the lymphocytes from participants with trichostatin A (TSA) for 24 hours and then measured changes in H3K9me2 relative to the control condition (dimethyl sulfoxide).. Patients with schizophrenia had significantly higher mean baseline levels of H3K9me2 than healthy controls (6.52 v. 2.78, p = 0.028). Moreover, there was a significant negative correlation between age at onset of illness and levels of H3K9me2 (Spearman's rho = -0.588, p = 0.008). In the lymphocyte cultures, TSA induced divergent responses in terms of H3K9me2 levels from patients with schizophrenia compared with healthy controls (F(1,14) = 5.082, p = 0.041).. The use of lymphocytes to study schizophrenia has its limitations because they may not be appropriate models of synaptic activity or other brain-specific activities.. Our results provide further evidence that schizophrenia is associated with a restrictive chromatin state that is also less modifiable using HDAC inhibitors.

Topics: Adult; Age of Onset; Blotting, Western; Cells, Cultured; Enzyme Inhibitors; Female; Histone Deacetylase Inhibitors; Histones; Humans; Hydroxamic Acids; Linear Models; Lymphocytes; Male; Methylation; Schizophrenia

Topics: Acetylation; Histone Deacetylase Inhibitors; Histone Deacetylases; Histones; Humans; Hydroxamic Acids; In Vitro Techniques; Lymphocytes; Schizophrenia