Joshua David

9 May 2015
English 363
A Review of the Effects of Epigenetic Modifications on a Schizophrenia Phenotype
Abstract
Schizophrenia is a severe brain disorder characterized by the development of psychosis.
It is understood that genetics has a huge role in developing schizophrenia. However, epigenetic
changes in the genome also determine expression. Methylation, an epigenetic modification, is
strongly correlated with a schizophrenic phenotype. The decreased gene expression caused by
methylation wreaks havoc on brain homeostasis, leading to psychosis. Knowing the relationship
between methylation and schizophrenia could pave the way to new gene therapies for the
disorder.
Introduction
Schizophrenia is a neurological disorder that is characterized by the manifestation of
psychosis, or disconnection from reality, in an individual. Psychosis affects the perception of an
individuals environment and, consequently, how that individual reacts to external stimuli.
Affected patients have a variety of symptoms, ranging from hallucinations and poor executive
function to motor movement disorders.
This debilitating brain disorder negatively impacts the wellbeing of people as well as the
economy. Schizophrenia afflicts about one percent of the global population (CDC, 2015). Of
those affected, 33% will attempt suicide and, ultimately, one in ten will succeed in taking their
own lives (CDC, 2015). In addition to self-harm, schizophrenic patients impose a financial
burden on the economy. In 2004, a study estimated the cost of healthcare and non-healthcare

expenses for schizophrenic patients to be 2.02 billion Canadian dollars (CDC, 2015). This debt
is increased by 4.83 billion dollars when accounting for unemployable Schizophrenic patients,
totaling to 6.85 billion dollars (CDC, 2015).
There are a multitude of factors that cause schizophrenia. Genetics play a major role in
phenotypic expression of schizophrenia. Schizophrenia aggregates in families, as it is a heritable
disease (Dempster et. al., 2010). Recent studies show that epigenetics heavily influences the
presence of psychosis as well. Epigenetics is the study of post-transcriptional changes to DNA
sequences that affect gene expression without altering the original sequence (Gavin et. al., 2010).
Several research articles link methylation, an epigenetic modification where methyl groups are
attached to a DNA sequence, with schizophrenia expression. Generally, methylation of DNA
sequences promotes a
heterochromatic state, seen in
Figure 1. Heterochromatin is
the formed when DNA folds on
itself to form a closed
configuration. Once closed,
transcription, and ultimately
gene expression, is inhibited
(Shmoop, 2008)

because cellular machinery

cannot access the closed gene.

Figure 1: DNA methylation effects at a chromosomal level. After methyl

groups are added to the DNA sequence, chromatin collapses and transcription
is reduced.

Epigenetic changes in discordant monozygotic twins

Dempster et. al. conducted a comprehensive genomic analysis of 22 monozygotic twins
that were discordant for schizophrenia, bipolar disorder, or both. This study highlights the role
of epigenetics in developing a schizophrenic phenotype as the two genomes are exactly the same.
Although disorders such as schizophrenia and bipolar disorder aggregate in families due to its
heritability, concordance in monozygotic twins for schizophrenia was not 100%. This trend
illustrates that non-genetic factors have a role in psychosis development.
Genome-wide epigenetic modifications, specifically DNA methylation, were measured in
the chromosomes of 22 twin pairs discordant for schizophrenia, bipolar disorder, or both.
Although the amount of methylation was similar in each monozygotic twin pair, the specific
CpG sites of methylation varied. CpG sites are guanine and cytosine rich areas of a DNA
sequence that are common targets of methylation. In addition to variability of methylation sites
between twin pairs, there was also variation in the degree of methylation of specific sites.
The most common methylation event Dempster et. al. found in affected twins was
hypermethylation upstream of the gene encoding pseudouridylate synthase 3 (PUS3).
Hypermethylation blocks transcription machinery from initiating transcription which leads to a
lack of expression. Another commonly affected CpG site in psychotic twins was
hypomethylation at ST6GALNAC1. ST6GALNAC1 is a sialytransferase molecule that
participates in cell-cell interactions (Dempster et. al., 2011). Its expression varies over the
course of development of the brain and central nervous system. When hypomethylated,
transcription is up-regulated and the gene is over-expressed. Since hypomethylation generally
leads to overexpression, it is hypothesized the disease could also arise from a duplication of this
gene.

There were several limitations with this experiment. Although this study had the highest
number of twin pairs in a disease-discordant MZ twin study, a larger sample size would give
stronger results. Another prevalent limitation is that peripheral blood was used to study the
genome rather than brain tissue. Evidence from other disorders show that epigenetic changes
may be tissue dependent (Dempster et. al., 2011). Finally, it is hard to show causality of
schizophrenia resulting from differently methylated regions in discordant twins. Since there was
no record of the subjects genomes before discordance for schizophrenia, there is no baseline for
comparison. A hypothesis is that these methylations may be caused by antipsychotic
medications, as the pathways in which these medications work are not clearly defined (Dempster
et. al., 2011).
Histone methylation
Histone methylation is a common example of an epigenetic modification. Specifically,
di-methylation at the 9th leucine on histone 3 (H3K9me2) results in a closed chromatin state
(Chase et. al., 2013). This heterochromatic state leads to reduced transcription levels. Since
schizophrenia has been previously linked with down-regulation of genes, Chase et. al.
hypothesized that schizophrenia can be identified by a restrictive epigenome.
To test this hypothesis, mRNA levels of GLP and G9a, histone methyltransferases
(HMTs) that di-methylate H3K9 into H3K9me2, were analyzed in postmortem brain tissue and
lymphocyte samples of schizophrenics, respectively (Chase et. al, 2013). Using RT-PCR,
increased levels of GLP mRNA were evident in schizophrenic patients compared to unaffected
controls. Similarly, increased levels of G9a mRNA were seen in lymphocytes of afflicted
patients versus controls.

GLP and G9a contribute to most of H3K9me2 modifications, which form

heterochromatic states (Chase et. al, 2013). H3K9me2 was the main focus of this experiment as
it encourages a closed genetic state. These higher levels HMTs lead to higher H3K9me2 levels,
and consequently, reduced transcription. Not only does methylation at H3K9 reduce gene
transcription at that histone, but it also acts as a site for more restrictive adaptors that lead to
more heterochromatic transformations (Huang et. al, 2007). This contagious behavior of closed
chromatin propagates a chain reaction that not only increases the risk of schizophrenia, but also
decreases the effectiveness of medications.
There are a couple of limitations that affect these results. Clinical symptoms were
correlated with peripheral tissue mRNA, not enzymes specific to synaptic function. However, it
is proposed that overall epigenetic regulation is not tissue specific (Chase et al. 2013). Another
limitation is that postmortem brain tissues only illustrate a snapshot of epigenetic changes at
death. Constant monitoring of peripheral blood cells is necessary to fully comprehend epigenetic
changes in schizophrenics.
DNA methylation of serotonin receptor 5HTR1A gene
Evidence also alludes to schizophrenic patients having a dysfunctional serotonin system
(Lesch, 1998). Serotonin is a biochemical neurotransmitter that regulates disposition and social
behavior, appetite, memory, sleep, and even libido. Many of these processes are impaired in
schizophrenics. Gray et. al. have identified a decrease in binding levels of 5HTR1A, a serotonin
receptor, as well as decreased 5HTR1A mRNA levels. A hypothesized cause of this decrease in
5HTR1A receptor binding levels is DNA methylation of the gene responsible for receptor
production (Carrard et al. 2011). When DNA is methylated at the gene promoter, transcription
can be inhibited.

Carrard et. al. tested this hypothesis by extracting DNA from peripheral blood leukocytes
in schizophrenics and assaying it using bisulfate sequencing and high-resolution melt assay.
Bisulfate sequencing converts unmethylated cytosines to uracil, but cannot convert methylated
cytosine. Sequencing is used after to identify modified 5HTR1A genes. They found an increase
in methylation percentages between the schizophrenic group and the healthy control group.
Methylation occurred at the gene promoter, which could block gene transcription. This blockage
could explain the lack of 5HTR1A receptors seen in schizophrenia patients and result in a
dysfunctional serotonin system (Tang et. al., 2014).
Conclusion
Although all these research articles observe varied levels of genomic structures such as
chromosomes, histones, and even individual genes, all of them show a link between methylation
and schizophrenia. Methylation instigates a closed chromatin state and subsequently inhibits
gene expression. Most of the genes targeted by methylation are necessary for proper brain
functionality and homeostasis. Disruption of these essential systems leads to psychosis, and
consequently, schizophrenia. With this knowledge, new therapies could be created in efforts to
combat schizophrenia. Schizophrenia is conventionally treated with antipsychotics, medication
that affects dopamine and serotonin neurotransmitters and receptors. Conversely, these
medications have severe but rare side effects such as increased risk of diabetes, heart disease,
stroke, and even motor movement disorders. These unintended repercussions may not be caused
by methylation-specific gene therapy. The precision of treating only affected methylation sites in
the genome may be more effective in individual cases than using antipsychotics that affect a
whole assortment of neurological processes. Hypothetical methylation therapies, however, can
only exist if more research is conducted.

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