EPIGENETIC MARKERS IN PERIPHERAL BLOOD AND LITHIUM RESPONSE IN PATIENTS WITH BIPOLAR DISORDER

M Zafrilla1, M Mitjans2,4, B Arias1,4, J Rodríguez, A Benabarre3,4

1Departament Biologia Evolutiva, Ecologia i Ciències Ambientals, Facultat de Biologia, IBUB, University of Barcelona, IBUB, Barcelona, Spain; 2Departament Genètica, Microbiologia i Estadística, Facultat de Biologia, IBUB, University of Barcelona, Barcelona, Spain; 3Bipolar Disorder Unit, Hospital Clínic, University of
Barcelona, IDIBAPS, Barcelona, Catalonia, Spain; 4Spanish Mental Health Research Network (CIBERSAM), Madrid, Spain.

INTRODUCTION RESULTS
Lithium (Li) is the first-line choice of long-term therapy for bipolar disorder (BD), due to its proven efficacy in both acute and DIFFERENTIALLY METHYLATED POSITIONS (DMPs)
maintenance phases1. Still, an adequate response may range from an excellent response in 24-45% to a complete lack of  A total of 130 CpG sites were significantly differentially methylated between Ex-Rp and N-Rp groups (p-value<8.72E-6;
response in 10-30% of patients2. Evidence from molecular biology has shown that Li exerts multiple effects on adjusted p-value<0.05) (Figures 1 and 2).
neurotransmitter/receptor-mediated signaling, ion transport, signal transduction cascades, hormonal, and circadian  From the genes mapped to these DMPs, FADS1, SORCS2, TRAF3IP2-AS1, MIR190, CACNA1B, and PDE2A have been
regulation. It also profoundly alters gene expression patterns with a final effect of stabilizing neuronal activities, supporting previously associated with BD, having been the last two also related to LR in addition to CYP1A1 13,14,15,16,17,18.
neural plasticity, and providing neuroprotection3,4.
In this sense, epigenetic mechanisms (such as DNA methylation) represent adaptive patterns of gene expression that might A. CpGs pattern in Ex-R group
result from and/or drive the effects of medications5. Recent studies reported that Li can modulate epigenetic mechanisms at
several levels of regulation. Thus, evidence suggests that pre-existing or Li-induced epigenetic mechanisms may play a role
56; 43%
in therapeutic response. 74; 57%

The aim of the present study is to compare genome-wide methylation patterns at a peripheral level between BD patients that
have had an excellent response to treatment with Li (Ex-Rp), and BD patients that are non-responders (N-Rp) in order to Hypermethylated Hypomethylated
detect specific differential methylated positions (DMPs) and differential methylation regions (DMRs). B. CpGs location
7; 5%

METHODS 26; 20%

65; 50%

SAMPLE 32; 25%

• 63 BD-I and BD-II unrelated Caucasian patients (42.31% females)(DMS-IV-TR criteria6). All patients gave their written
Open sea Island Shore Shelf
informed consent, and approval from each institution's ethics committees was obtained.
C. CpGs mapped to genes
RESPONSE TO LITHIUM
Two groups of individuals were selected based on their LR7*: 37; 28%

• N-Rp group (n=37): BD patientEx-Rp group (n=26): BD patients presenting a 50% reduction of the episodes after the 93; 72%
introduction of Li in monotherapy.
• s who did not reduce at least 50% of the episodes and patients who required electroconvulsive therapy. Mapped to genes Not mapped to genes
*No differences were found for age (p=0.321) and sex (p=0.888). Figure 1. (A) Summary of the 130 CpG sites that
are differentially methylated in Ex-Rp compared
METHYLATION ANALYSES to N-Rp (%). (B) Location of CpGs in the genome
Figure 2. Differentially methylated CpG sites between Ex-Rp and N-Rp BD patients. The genes associated with the
Top20 most significant CpG sites appear annotated above each differentially methylated CpG site.
• DNA extractions from peripheral-blood samples from each participant were extracted according to standard protocols. (%). (C) CpG sites mapped to genes (%).

• Methylation profiling was obtained using Infinitum HumanMethylationEPIC BeadChip Kit (Illumina) in CeGen-ISCIII. DIFFERENTIALLY METHYLATED REGIONS (DMRs)
• Benjamini-Hochberg multiple-testing correction was used to correct the false discovery rate (FDR) and a p-value of 0.05  We found 16 DMRs between Ex-Rp and N-Rp groups (FDR adjusted p-value<1.48E-10): 9 were hypomethylated and 7
was considered for significance. hypermethylated in Ex-Rp compared to N-Rp.
 Two of the DMRs were linked to genes previously related to BD, and both were hypomethylated in Ex-Rp (Figure 3): Firstly,
Methylation data proces s ing Covariates es timated Differentially Methylated Positions (DMPs) DDR1, in DMR219, (Mean difference= -0.035), and secondly ANK3, in DMR8 (Mean difference= -0.04), which also was
from methylation data previously associated with LR20.
ChAMP R package 8,9 Limma R package11
Blood cell type
Raw IDAT File Import proportion
es timates .
Probe Filtering Refbase

Smoking s core. Differentially Methylated Regions (DMRs)

Normalization EpiSmokEr 10 Figure 3. Differentially methylated regions between Ex-Rp and N-Rp associated with genes previously related to BD.
DMRcate R package12
Batch Effect CONCLUSIONS
Correction
Significant differences in methylation for 130 CpG sites and 16 regions were found between Ex-Rp and N-Rp. Some of the
genes related to both DMPs and DMRs have been consistently associated with BD and LR, among other psychiatric
M- value Matrix
disorders.
742,902 probes
These results point to the importance of considering epigenetic markers when studying LR. Further research is needed for a
CONTACT: marinazafrilla9@gmail.com better understanding of LR and, ultimately, help in the better choice of proper treatment for BD patients, thus moving
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