Poster

Poster 6 Mood Disorders 1 ポスター 6 気分障害1

P6-1 Development of diagnostic method for postpartum depression using comprehensive DNA methylation analysis 網羅的メチル化解析による産後うつ病の診断法開発 Nakamura Yukako（中村由嘉子）¹，中杤昌弘²，國本正子¹，岡田俊³，Aleksic Branko¹，遠山美穂¹，森川真子³，山内彩¹，大原聖子¹，椎野智子¹，久保田智香¹，佐藤真耶¹，安藤昌彦²，尾崎紀夫^1,3 ¹Department of Psychiatry, Nagoya University Graduate School of Medicine ²Statistical Analysis Section, Center for Advance Medicine and Clinical Research, Nagoya University Hospital ³Center for Advanced Medicine and Clinical Research, Nagoya University Hospital Background　It is pointed out that epigenetic modifications affected by environmental factors may closely be related to depression, and DNA methylation(DNAm) may be used as a biomarker for diagnosis of depression. Methods　In this study, we conducted comprehensive methylation analysis using peripheral blood of women who are expected to deliver baby with the aim of developing a diagnostic method for postpartum depression (PPD). A total of 18 female cases and 18 female controls were enrolled. Genome wide DNAm profiles were obtained by analysis with an Infinium Human Methylation 450BeadChip. We performed a case-control study to examine the association of DNAm status (p <1.3×10 ^-7). After that we conducted functional enrichment analysis of PPD through the Database for Annotation, Visualization and Integrated Discovery (DAVID) to explore enriched functional-related gene group of PPD. The study protocol was approved by the Ethics Committee of the Nagoya University Graduate School of Medicine. Written informed consent was obtained from all individual participants included in the study. Result and discussion　We observed no genome-wide significant association between the genome-wide measure of DNAm based on all CpG sites and PPD. From a result of comparing the case-control DNAm level, we selected 1,000 sites with low p-values and analyzed by DAVID6.8. As a result of analysis done using DAVID 6.8, we found four GO Terms (protein binding, nucleoplasm, transcription corepressor activity, axon guidance) with false discovery rate < 0.05. From the results of this study, we confirmed that not only a single methylation site but also an analysis focusing on their functional relevance was useful for development of a diagnostic method for postpartum depression.		P6-2 Relation of the DGKH genotype with openness to experience, a premorbid personality trait of bipolar disorder DGKH遺伝型の双極性障害の病前人格である開放性との関係 Matsumoto Yoshihiko（松本祥彦）¹，鈴木昭仁¹，白田稔則¹，高橋奈那¹，能登契介¹，後藤薫²，大谷浩一¹ ¹Department of Psychiatry, Yamagata University School of Medicine, Yamagata, Japan ²Department of Anatomy and Cell Biology, Yamagata University School of Medicine, Yamagata, Japan Aims: The diacylglycerol kinase η gene (DGKH) is one of the few replicated risk genes for bipolar disorder. Meanwhile, specific personality traits, especially openness to experience, have been reported as premorbid personality traits of the disorder. In the present study, we examined the relation of the DGKH genotype with broad dimensions of personality, to obtain further evidence for its implication in the etiology of bipolar disorder.Methods: The subjects were 319 healthy Japanese. Personality assessment was performed by the NEO Personality Inventory-Revised, which has the neuroticism, extraversion, openness to experience, agreeableness and conscientiousness dimensions. The A/G polymorphism of DGKH (rs9525580) was identified by a PCR-RFLP method. The subjects were divided into two groups with respect to the presence or absence of the A allele, which is a putative risk allele for bipolar disorder. The Ethics Committee of Yamagata University School of Medicine approved the study protocol, and all subjects provided written informed consent to participate.Results: The group with the A allele had higher scores of openness to experience compared to that without this allele. The DGKH genotype did not influence scores of other dimensions.Conclusions: The present study shows that a bipolar-risk allele of DGKH is associated with higher openness to experience, providing further evidence for the implication of this gene in the etiology of bipolar disorder.Keywords: Diacylglycerol kinase; Polymorphism; Personality; Openness to experience

P6-3 The alternation of the splicing patterns caused by a de novo mutation of UNC13B found in a patient with bipolar disorder 双極性障害患者におけるUNC13Bのデノボ変異がもたらすスプライシングパターンの変化 Jimbo Kotori（神保ことり）^1,2，中村匠^2,3，坪井貴司³ ¹Faculty of Medicine, Nara Medical University, Nara, Japan ²Laboratory for Molecular Dynamics of Mental Disorders, RIKEN Center for Brain Science, Saitama, Japan ³Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, Tokyo, Japan Bipolar disorder is one of the mental disorders and characterized by manic and depressive episodes. It is difficult to be diagnosed, and some patients have intractable bipolar disorder because of the long term misdiagonsis, so that the development of accurate diagnoses is urgently needed. However, the mechanisms are still unclear, although the importance of the genetic factor is well known. We identified 9 de novo loss-of-function mutations in 79 trios by the previously performed trio-based exome sequencing. Among them, a mutation of UNC13B, which regulates synaptic priming, is the only one which is predicted to change the splicing patterns (Kataoka M, Matoba N et al. Molecular Psychiatry. 2016). The mutation of UNC13B is located on the donor site of exon39. In this report, we amplified exon38-40 of UNC13B, inserted into pAcGFP-C1 vector, and performed splicing assay. As a result, an exon39-skipping variant was significantly increased by the mutation. This variant is predicted to escape nonsense-mediated mRNA decay and causes a truncated protein losing the latter half of the C2C domain which is important for priming. This truncated protein might cause abnormal neurotransmission by dominant negative effect, because UNC13B works as a homodimer. In summary, we found that the de novo mutation of UNC13B caused the exon-skipping variant, and that the abnormal synaptic priming might be occurred in the patient’s organisms.		P6-4 Effects of return-to-work program on autonomic nervous system activity in workers on sick leave due to depression リワークプログラムが大うつ病性障害・双極性障害の患者の自律神経活動へもたらす影響について Hattori Saki（服部早紀）¹，須田顕¹，岸田郁子^1,2，宮内雅利¹，白石洋子^1,3，藤林真美⁴，辻田那月⁵，石井千恵²，石井紀夫²，森谷敏夫⁶，平安良雄^1,7 ¹Department of Psychiatry, Yokohama City University School of Medicine, Yokohama, Japan ²Fujisawa Hospital, Fujisawa, Japan ³Department of clinical laboratroy, Yokohama City University Hospital, Yokohama, Japan ⁴Setsunan Univrsity, Neyagawa, Japan ⁵Graduate School of Human and Environmental Studies, Kyoto University, Kyoto, Japan ⁶Faculty of General Education, Kyoto Sangyo University, Kyoto, Japan ⁷Heian Hospital, Urasoe, Japan Sick leave with mental disorders, such as major depressive disorder or bipolar disorder, has increased in Japan, and return-to-work (RTW) program have been executed in recent years. However, the biological effect of RTW program is unclear yet. The aim of this study was to evaluate the effect of RTW program on autonomic nervous system (ANS) and psychiatric symptoms of the workers on sick leave due to mental disorders. Subjects were 104 Japanese workers on sick leave due to major depressive disorder or bipolar disorder. They received RTW program for 3 months in Yokohama City University Hospital (YCUH). The ANS activity was evaluated using heart rate variability (HRV) power spectral analysis at the beginning and ending of 3 months program. Psychiatric symptoms were evaluated by MADRAS-J and SASS. The institutional ethics committee of YCUH approved the study protocol, and all participants signed written informed consent statements. Parasympathetic activity at the ending was significantly higher than that at the beginning (p=0.014). Psychiatric symptoms were significantly improved at the ending (p<0.001). We could follow up 88 participants 3-months after the program, and 60 participants returned to work. The beginning and ending ANS activities were not significantly different between the patients who could return to work and patients who could not. In conclusion, RTW program is effective for improving both parasympathetic activity and psychiatric symptoms in workers on sick leave due to major depressive disorder or bipolar disorder. However, ANS activities were not enough as predictive indicators of RTW. Various occupational factors could be associated with RTW. Further studies for larger sample size are needed to clarify the biological effect of RTW program.

P6-5 Carbonyl stress in mood disorder 気分障害とカルボニルストレス Miyashita Mitsuhiro（宮下光弘）^1,2,3,5，鈴木一浩^1,2,3,5，堀内泰江¹，鳥海和也¹，江越正敏^1,2，今井淳司²，長瀬幸弘³，湯澤公子⁴，瀧澤俊也⁴，鷲塚伸介⁵，糸川昌成^1,2，新井誠¹ ¹Project for Schizophrenia Research, Tokyo Metropolitan Institute of Medical Science, Tokyo, Japan ²Department of Psychiatry, Tokyo Metropolitan Matsuzawa Hospital, Tokyo, Japan ³Takatsuki Clinic, Akishima, Japan ⁴Department of Neurology, Tokai University School of Medicine, Isehara, Japan. ⁵Department of Psychiatry, Shinshu University School of Medicine, Matsumoto, Japan 【目的】我々はこれまでに、統合失調症の約2割において、終末糖化産物であるペントシジンが蓄積しビタミンB6が枯渇するという、カルボニルストレスの亢進、を明らかにした。しかしながら、他の精神疾患におけるカルボニルストレスの存否は現在までに報告が無い。そこで、本研究では気分障害を対象に、カルボニルストレス関連分子である血漿中ペントシジン濃度および血清ビタミンB6濃度を測定し、気分障害におけるカルボニルストレスの分布を明らかにすることを目的とした。【方法】当研究室で保有しているうつ病、双極性障害の末梢血を使用して、high performance liquid chromatography（HPLC）により血漿ペントシジン濃度を測定した。また、ビタミンB6濃度はSRL社に委託して定量した。統合失調症および健常群は、2012年9月以降のサンプルを使用した。得られたデータは、GraphPad PRISM v6.0で解析を行った。【結果】統合失調症のみならず、躁うつ病、うつ病患者においてもカルボニルストレスが亢進している患者を認めた。しかしながら、血漿ペントシジン高値（55.2ng/ml以上）かつビタミンB6低値を、カルボニルストレス亢進、と定義した場合、気分障害でカルボニルストレスが亢進する患者の割合は、統合失調症と比較して低かった。また、躁うつ病はうつ病と比較して、カルボニルストレスが亢進する患者が多いことが示された。【考察】カルボニルストレスの亢進は気分障害においても存在するが、その程度は統合失調症より軽いものであった。発表当日は、交絡因子の調整および臨床データを踏まえながら考察する。		P6-6 Oxidative stress and mitochondrial defects in the paraventricular thalamic nucleus of patients with bipolar disorder 双極性障害患者死後脳を用いた視床室傍核における酸化ストレス、およびミトコンドリア異常の探索 Kubota-Sakashita Mie（窪田-坂下美恵）¹，磯野蕗子¹，Turecki Gustavo²，Mechawar Naguib²，加藤忠史　¹ ¹Lab for MDMD, RIKEN CBS, Saitama, Japan ²Douglas Mental Health Institute, McGill University, Quebec, Canada To investigate a possible link between bipolar disorder (BD) and mitochondrial dysfunction, we have explored the cells with mitochondrial dysfunction in formalin-fixed paraffin embedded (FFPE) postmortem brain samples of patients with BD. Based on the qPCR analysis to detect mitochondrial DNA (mtDNA) deletions In the brain of transgenic (Tg) mice expressing mutant mtDNA polymerase (polymerase γ; Polg1), we focused on the paraventricular thalamic nucleus (PVT). By double staining method using antibodies against the cytochrome c oxidase (COX), a mtDNA-encoded protein, and succinate dehydrogenase (SDH), a nuclear-encoded mitochondrial protein, we could confirm that the COX-negative cells in PVT accumulated mtDNA deletions in the Tg mice by a laser micro dissection technique. It suggests that the COX-negative cells resulted from mtDNA deletions. Thus, we searched for the cells with mitochondrial dysfunction in FFPE sections of the thalamus (9 BD patients and 9 control subjects). The COX-negative cells were found in a number of samples derived from patients with BD. However, some of control samples with low pH also showed COX-negative-like cells. To discriminate the results from ante-mortem and post-mortem artifacts, we tried to immunohistochemical staining for the oxidative stress marker, 8-hydroxy- 2'-deoxyguanosine (8-OHdG), because oxidative stress may be involved in the pathophysiology of BD. A numbers of 8-OHdG positive cells were significantly higher in patients with BD. The signals were localized in area where the COX-negative cells were frequently found, such as PVT. This suggests that the PVT neurons are susceptible to the oxidative stress, and it may be causative of the mitochondrial dysfunction in BD.

P6-7 Estimation of depression-like behaviors using a test battery in drug-induced model of common marmosets マーモセットの薬物惹起性モデルにおけるテストバッテリーを用いたうつ様行動の評価 Yamanaka Hajime（山中創）¹，石橋英俊²，高田昌彦¹，中村克樹³ ¹Systems Neuroscience, Primate Research Institute, Kyoto University, Inuyama, Japan ²Pre-clinical Research Center, Tokyo Medical University, Tokyo, Japan ³Cognitive Neuroscience, Primate Research Institute, Kyoto University, Inuyama, Japan Depression is a serious mood disorder with a high prevalence. Although research works on depression have so far been performed mainly in rodents, the use of nonhuman primates is considered to improve the efficiency of assessing the similarity of depressive symptoms, termed face validity. Common marmosets have increasingly been utilized in recent years as potential models of neurological/psychiatric disorders. Since diagnostic criteria for psychiatric disorders are primarily based on altered behavioral types, the development of an appropriate test battery is critical to assess depression-like behaviors. In this study, several behavioral tests were executed to validate their utility in marmosets. Then, a battery of six tests were established through repeated measurements over several days in a reserpine-induced depression model. First, a peephole test was developed to examine visual exploratory behavior (VEB) that is frequently discussed in terms of motivation, curiosity, and interest. We found that conspecific individuals used as presentation stimulus effectively induced VEB. Second, a sucrose preference test was applied for investigating anhedonic behavior. In this test, a sucrose concentration-response curve was generated to reveal that 1.8% sucrose was optimal in marmosets. The reserpine-induced response patterns were tested using a battery with four more tests including food intake. Of the nine marmosets, one was identified to exhibit typical response patterns. The incidence rate (11%) in our study was nearly equivalent to the clinical incidence rate (18%) for depressive patients with reserpine treatment. Thus, the present test battery might have a potential to assess the face validity of depression-like behaviors in primate models.