Poster

Poster 5 Schizophrenia 1 ポスター 5 統合失調症1

P5-1 Developing a method for detection of novel LINE-1 insertion using single brain cells ヒト死後脳を使用したシングルセルレベルにおけるLINE-1新規挿入部位の決定 Bundo Miki（文東美紀）^1,2，上田順子³，西岡正樹³，清田恵美¹，笠井清登⁴，加藤忠史³，岩本和也¹ ¹Molecular Brain Science, Kumamoto Univ. Kumamoto, Japan ²PRESTO JST, Saitama, Japan ³Lab for Molecular Dynamics of Mental Disorders, RIKEN BSI, Saitama, Japan ⁴Dept Neuropsy, Univ of Tokyo, Tokyo, Japan Recent several studies have unveiled that the genome of brain cells contains various somatic mutations, such as single nucleotide variants, copy number variations and novel insertions of retrotransposons. These brain cell-specific somatic mutations have the potential to be associated with the onset of psychiatric diseases. We recently reported that the copy number of long interspersed nucleotide element (LINE-1) retrotransposon was higher in the neurons of patients with schizophrenia than in the neurons of healthy controls. The purpose of this study is to detect the somatic insertion sites of LINE-1 using human postmortem brains at the single cell level.Here we developed a new method to identify the brain-specific LINE-1 insertion genomic site at the single cell level. Firstly, cell nuclei of several specific brain cell types were separated from human prefrontal cortex using nuclei sorting method. Whole genome amplification (WGA) using single nuclei was performed and WGA products with low allele dropout rate were selected. Genomic regions including 3’ end of human LINE-1 (L1Hs) and adjacent regions were selectively amplified by adaptor-ligation PCR and sequenced by illumina MiSeq. We detected 79.2% (670/859) of LINE-1 found in the reference human genome sequence (hg38) and 64-74 of unknown non-reference somatic LINE-1 insertions in single neuronal nuclei from prefrontal cortex of a human without any psychiatric diseases. These newly inserted LINE-1 were validated by Sanger sequencing method. Using the developed method, we have been conducting a case-control study using postmortem brain samples from patients with schizophrenia and controls to clarify the role of somatic LINE-1 insertions in the pathogenesis of schizophrenia.		P5-2 Establishment of a new protocol for quantifying the copy number of active LINE-1 subfamilies in mice 転移活性を有するマウスLINE-1のコピー数定量のための新規プロトコール確立 Kuroki Ryota（黒木遼太），村田唯，澤村理英，文東美紀，岩本和也 Department of Molecular Brain Science, Graduate School of Medical Sciences, Kumamoto University, Kumamoto, Japan LINE-1 is a retrotransposon that can increase its copy number by the process called retrotransposition. The new LINE-1 copy can be inserted into other genomic regions, thereby affecting stability of the genome and regulation of expression from neighboring genes. Recently, we found increased LINE-1 copy number in brains of schizophrenic patients, and in the brains of mice born from poly (I:C) injected dam (Bundo et al., 2014). The structure and evolutionary characteristics of LINE-1 differ between humans and mice. In humans, only the most evolutionarily youngest LINE-1 subfamily, Hs, retains retrotransposition activity, while three subfamilies (A, Gf, Tf) retain activity in mice. In addition, in mice LINE-1, repeat sequences called monomers are present in the 5'-UTR, but this structure is not found in humans. Therefore, to study the LINE-1 dynamics in mice in detail, specific attentions should be required. Here we established the quantification protocol of copy number of each of three active LINE-1 subfamily in mice. We retrieved total of 34 mice LINE-1 consensus sequences from Repbase, and designed the PCR primer pairs so that 3' ends were unique to the consensus sequence of the target subfamily. We then chose primer pairs that satisfied the following criteria: 1) single band product with the expected size, 2) stable amplification curve and unimodal dissociation peak by quantitative PCR, 3) low background by direct sequencing of the PCR product, 4) high specificity by cloning-sequencing analysis of the PCR product. As a result, we successfully obtained at least one primer pair for each active subfamily. Results of ongoing copy number estimation in various mice models of psychiatric disorder will also be presented.

P5-3 Glutamatergic Neurometabolite Levels in Patients with Ultra Treatment-Resistant Schizophrenia: a Cross-sectional 3T Proton MRS study Nakajima Shinichiro（中島振一郎）^1,2，岩田祐輔^1,2，プリッとマンエリック²，カラヴァッジョフェルナンド²，キムジュリア²，シャーパリータ²，マーワナ²，ゲレッツェンフィリップ²，チャベスソフィア²，デルーカヴィンチェンツォ²，三村將²，レミントンゲイリー²，グラフアリエル² ¹Department of Neuropsychiatry, School of Medicine, Keio University, Tokyo, Japan ²Toronto, Canada Objective: In terms of antipsychotic treatment response, patients with schizophrenia can be classified into three groups: (1) treatment-resistant to both non-clozapine (CLZ) antipsychotics and CLZ (ultra treatment-resistant schizophrenia [URS]), (2) treatment-resistant to non-CLZ antipsychotics but CLZ-responsive schizophrenia [non-URS]), and (3) responsive to first-line antipsychotics (treatment non-resistant schizophrenia [TnRS]). This study aimed to compare glutamatergic neurometabolite levels among these three patient groups and healthy controls (HCs), using proton magnetic resonance spectroscopy (1H-MRS).Method: Glutamate (Glu) and glutamate+glutamine (Glx) levels were assessed in the caudate, the anterior cingulate cortex (ACC), and the dorsolateral prefrontal cortex (DLPFC) using 3T 1H-MRS (PRESS, TE=35ms). Glutamatergic neurometabolite levels were compared between the groups using analyses of variance. This research was approved by REB of CAMH. Results: A total of 100 participants were included, which consisted of 26 patients with URS, 27 patients with non-URS, 21 patients with TnRS, and 26 HCs. Group differences were detected in ACC Glx levels (F[3,96]=2.93, p=0.038); patients with URS showed higher ACC Glx levels than HCs (p=0.038). There were no group differences in the caudate or DLPFC. When patients with URS and non-URS were combined into one group as treatment-resistant schizophrenia patients, this subset of patients showed higher Glu and Glx levels in the ACC compared to HCs (p=0.028 and 0.023, respectively).Conclusions: Higher levels of glutamatergic neurometabolites in the ACC may be a trait marker of treatment-resistant schizophrenia, which may become prominent after initial antipsychotic treatment failure and persist even after CLZ administration.		P5-4 Smoking rates and number of cigarettes smoked per day in schizophrenia: A large cohort meta-analysis in a Japanese population 日本人統合失調症における喫煙率および喫煙本数: 大規模メタ解析 Ohi Kazutaka（大井一高）^1,2，嶋田貴充¹，桑田有紀¹，片岡譲¹，大久保裕章¹，記村康平¹，康山俊樹¹，上原隆¹，川崎康弘¹ ¹Dept. of Neuropsychiatry, Kanazawa Med. Univ. ²Medical Research Institute, Kanazawa Med. Univ. Background: Cigarette smoking is consistently more common among schizophrenia patients (SZ) than general population (GP) worldwide; however, the findings of studies in Japan are inconsistent. Recently, the smoking rate has gradually decreased among GP. Methods: We performed a meta-analysis of smoking status in a large Japanese cohort of (i) 1,845 SZ and 196,845 GP and (ii) 842 SZ and 766 psychiatrically healthy controls (HCs) from twelve studies over a 25-year period, including 301 patients and 131 controls from our study. Results: In our case-control sample, SZ had a significantly higher smoking rate than HCs (p=0.039). The proportion of heavy smokers (p=0.027) and the number of cigarettes smoked per day (CPD, p=8.20×10-3) were significantly higher among SZ than HCs. For the smokers in SZ, atypical antipsychotics dosage was positively correlated with CPD (p=1.00×10-3). A meta-analysis found that SZ had a higher smoking rate than GP for both men [odds ratio (OR)=1.53, p=0.035, SZ_52.9%, GP_40.1%] and women (OR=2.40, p=1.08×10-5, SZ_24.4%, GP_11.8%). In addition, male SZ had a higher smoking rate than male HCs (OR=2.84, p=9.48×10-3, SZ_53.6%, HC_32.9%), but the difference was not significant for women (OR=1.36, p=0.53, SZ_17.0%, HC_14.1%). Among both males and females, SZ had a higher smoking rate than both GP (OR=1.88, p=2.60×10-5) and HCs (OR=2.05, p=0.018). These rates were not affected by the patients’ recruitment year (p>0.05). The CPD values of SZ and GP were 22.0 and 18.8, respectively. Conclusions: SZ are approximately 2 times more likely to smoke than GP and HCs based on data collected over a decade in Japan.

P5-5 Characteristics of resting-state EEG power spectrum in patients with chronic schizophrenia 慢性統合失調症における安静時脳波の特性 Miyazaki Takahiro（宮崎貴浩）^1,2，垂水良介¹，中島振一郎^1,5，和田真孝¹，本多栞³，藤井進也⁴，増田史¹，三村將¹，野田賀大¹ ¹Department of Neuropsychiatry, Keio University, Tokyo, Japan ²Nishigahara Hospital, Tokyo, Japan ³Graduate School of Media and Governance, Keio University, Tokyo, Japan ⁴Faculty of Environment and Information Studies, Keio University ⁵Centre for Addiction and Mental Health, Department of Psychiatry, University of Toronto Background: Limitations in both the efficacy and response rate of currently available treatments for schizophrenia represent a significant unmet need for both the affected patients and the society. Revealing the underlying pathophysiological mechanisms of this disorder will leverage the quest for novel therapeutic options. Previous research has demonstrated altered spontaneous EEG activities in patients with schizophrenia. In the present study, we further explored to clarify the relationships among EEG activities, the clinical severity, and medication response of schizophrenia.Methods: Forty-seven patients with chronic schizophrenia were included. Severity of the remaining symptoms was assessed with Positive and Negative Syndrome Scale (PANSS). Resting-state EEG was collected from the 19 electrodes with the international 10-20 system. The EEG data were pre-processed and 2-second epochs were extracted. The epoched data were band-pass filtered at delta, theta, alpha, beta, and gamma frequencies and Hilbert-transformed to obtain the amplitude values for each frequency band. The effect of the PANSS score on root-mean-squared amplitudes for each frequency bin was tested with the linear regression with the smoking history and chlorpromazine equivalent dosage as covariates.Results: The delta (p = 0.036) and theta (p = 0.024) amplitudes were significantly larger for higher PANSS score in this population.Discussion: Despite the limited statistical power of the study due to the small number of participants, the results showed the possible link between the symptomatic severity of schizophrenia and resting-state EEG-based physiological measures. Also, the results suggested need for further studies on the spontaneous EEG activities related to treatment resistance.		P5-6 Suppression of Centaurin gamma 1A rescues the reduced prepulse inhibition in Drosophila larval model of fragile X syndrome センタウリンγ1Aの発現抑制は脆弱性X症候群モデルハエ幼虫におけるプレパルスインヒビション減少を回復させる Morimoto Takako（森本高子），松本悠太郎，山内淳司 Lab of Neuroscience and Neurology, Sch of Life Sci, Tokyo Univ of Pharm and Life Sci The neural mechanisms of the psychiatric disorder like autism spectrum disorder and schizophrenia has been intensively studied and a number of genes have been identified as candidates. However, the inclusive understanding of relation between the gene and the function of neural system has not been clarified yet. In the last meeting, we reported a prepulse inhibition (PPI) phenomenon by using Drosophila larval hearing. PPI is a neurological phenomenon found in humans and other organisms and using for the diagnostic test of schizophrenia as well as autism. Our finding is the first report of PPI in Drosophila. Here, we have examined the PPI in two mutants, fmr1 and centaurin gamma 1A (CenG1A), which are supposed to relate the psychiatric disorder. fmr1 is a responsible gene of fragile x syndrome and codes for a protein called the Fragile X mental retardation protein (FMRP). CenG1A has GTPase, PH, ArfGAP and ANK domains. In addition, it has been reported that one of centaurin family members is included in the chromosomal deletion region found in autistic patients. Further, we revealed its potential function as a negative regulator of neurotransmitter release (Homma, et al, 2014). We found that either fmr1 homozygotes or CenG1A heterozygotes showed reduced PPI. CenG1A homozygotes almost died before 3rd instar larval stage. It has been suggested that CenG1A mRNA is associated with FMRP, leading to increased CenG1A protein levels in fmr1 knockout mice. Recent report showed that genetic reduction of CenG1A rescued the phenotype in FXS flies. Therefore, we examined and successively observed PPI in FXS larvae with genetically reduced expression of CenG1A, supporting the hypothesis. Thus, the expression level of CenG1A is one of possible key factors for psychiatric defects.

P5-7 Predictive Accuracy for Work Outcome in Patients with Schizophrenia: Examination Based on the Functioning Levels 統合失調症における労働状態の予測の精度:機能度による検討 Sumiyoshi Chika（住吉チカ）¹，藤野陽生²，山森英長³，工藤紀子⁴，畦地裕統⁴，藤本美智子³，安田由華³，大井一高⁵，住吉太幹⁶，橋本亮太^3,4,7 ¹Faculty of Human Development and Culture, Fukushima University, Fukushima, Fukushima, Japan ²Department of Special Needs Education, Oita University, Oita, Oita, Japan ³Department of Psychiatry, Osaka University Graduate School of Medicine, Suita, Osaka, Japan ⁴Molecular Research Center for Children's Mental Development, United Graduate School of Child Development, Osaka University, Suita, Osaka, Japan ⁵Department of Neuropsychiatry, Kanazawa Medical University, Uchinada, Ishikawa, Japan ⁶Department of Preventive Intervention for Psychiatric Disorders, National Institute of Mental Health, National Center of Neurology and Psychiatry, Kodira, Tokyo, Japan ⁷Department of Pathology of Mental Diseases, National Institute of Mental Health, National Center of Neurology and Psychiatry, Kodira, Tokyo, Japan [Background] We previously reported that intelligence decline, social function, and psychiatric symptoms were prominent predictive factors for work outcome in patients with schizophrenia. The purpose of this study was to examine the accuracy of the prediction based on functioning levels of patients. [Methods] Subjects: One-hundred and thirty-five Japanese patients meeting DSM-IV-TR criteria for schizophrenia entered the study. The study was approved by the Ethics Committee of Osaka University. Analyses: Four separate multiple logistic regression analyses were conducted to predict work outcomes which were dichotomized by the criterion of 0, 10, 20, or 30 work hours per week. Probabilities for exceeding each criterion (estimated outcomes) were calculated using predictive factors that remained significant in regression models (intelligence decline, psychiatric symptoms, and social function). High- (poor-) functioning patients were defined as those who were in better conditions as to the predictive factors. Frequency distributions for the two groups were produced to examine the discrepancy between estimated and observed outcomes. [Results] Overall, prediction was less accurate in high-functioning patients; the distributions were almost uniform in all criteria. Also, substantial portion of high-functioning patients demonstrated poor observed outcomes. [Discussion] The results indicated that predictive accuracy in work outcome was less reliable in relatively high-functioning patients with schizophrenia. Extrinsic variables, such as local economy or welfare services, may disturb the translation from functional capacity to work outcome. In addition, some of high-functioning patients may have better insight into their work capacity, which may refrain from engaging in long-hour work.		P5-8 Abnormalities of eye movement are associated with work hours in schizophrenia 統合失調症における眼球運動と労働時間の関連 Morita Kentaro（森田健太郎）¹，三浦健一郎²，藤本美智子³，宍戸恵美子⁴，椎野智子⁴，高橋潤一⁵，山森秀長³，安田由華³，工藤紀子⁶，平野羊嗣⁵，越山太輔¹，岡田直大^1,7，池田学³，鬼塚俊明⁵，尾崎紀夫⁴，笠井清登^1,7，橋本亮太^3,6,8 ¹Department of Neuropsychiatry, Graduate School of Medicine, University of Tokyo, Tokyo, Japan ²Department of Integrative Brain Science, Graduate School of Medicine, Kyoto University, Kyoto, Japan ³Department of Psychiatry, Osaka University Graduate School of Medicine, Osaka, Japan ⁴Department of Psychiatry, Nagoya University Graduate School of Medicine, Nagoya, Japan ⁵Department of Neuropsychiatry, Graduate School of Medical Sciences, Kyushu University, Fukuoka, Japan ⁶Molecular Research Center for Children's Mental Development, United Graduate School of Child Development, Osaka University, Osaka, Japan ⁷The International Research Center for Neurointelligence (WPI-IRCN) at The University of Tokyo Institutes for Advanced Study (UTIAS), Tokyo, Japan ⁸Department of Pathology of Mental Diseases, National Institute of Mental Health, National Center of Neurology and Psychiatry, Kodaira, Tokyo, Japan Eye movement abnormalities have been reported in schizophrenia; however, their influences on everyday life and social functioning still remain unknown. In this study, we explored the association between eye movement and actual work hours.By analyzing eye movement data gathered from a sample of 69 subjects with schizophrenia and 246 healthy subjects, we found positive correlations between eye movement measures and work hours per week, which were only significant in subjects with schizophrenia. We also conducted a replication study in 118 subjects with schizophrenia and 280 healthy subjects from four institutions (Osaka University, University of Tokyo, Nagoya University, and Kyushu University). The findings of positive correlations between the eye movement score and work hours, which were only significant in subjects with schizophrenia, were confirmed in this additional dataset as well.Our results revealed an association between eye movement and work hours in schizophrenia. This association is unsurprising in the context of previous literature because eye movement abnormalities have been shown to be associated with neurocognitive measures requiring visual processing, and studies have shown that visual processing, mediated by social functioning or negative symptoms, affects real-world functioning. There is still much need to elucidate how eye movement is associated with work hours; however, this novel finding of a neurophysiological measure such as eye movement is associated with clinical recovery in schizophrenia could lead to fruitful findings in future research.

P5-9 A preliminarily study of near-infrared spectroscopy to measure a resting state activity in prefrontal cortex of schizophrenia 統合失調症におけるNIRSを用いた安静時脳血流測定のための予備的研究 Yanagi Masaya（柳雅也）¹，細見史治¹，川久保善宏¹，土屋有希¹，廣瀬智之¹，三川和歌子¹，辻井農亜¹，尾崎哲²，白川治¹ ¹Department of Neuropsychiatry, Kindai University Faculty of Medicine, Osaka-sayama, Osaka, Japan ²Izumigaoka Hospital Introduction) Among neuroimaging modalities, near-infrared spectroscopy (NIRS) has an advantage to be convenient and safe, which is useful for clinical application, for measuring blood flow changes. We previously presented a poster in the 39^th Annual Meeting of Japanese Society of Biological Psychiatry that show a measurement of resting-state activity using NIRS in healthy subjects. In this study, the NIRS measurement of resting state activity was applied for patients with schizophrenia to show the utility for further application to clinical research of schizophrenia. Methods)NIRS measurement was performed using a 10-channel NIRS device to detect blood flow changes in prefrontal cortex, and low pass filter (<0.1Hz) was applied for the blood flow fluctuations. 20 male patients with schizophrenia and case-matched 20 control subjects participated in this study. Written informed consent was obtained from all subjects. The current study was approved by the Ethics Committee of Kindai University Faculty of Medicine. Results)The magnitude of the blood flow fluctuations was reduced in mPFC in patients with schizophrenia compared to the controls, which is similar with the findings in fMRI study that show a reduced amplitude of low frequency fluctuations in mPFC in schizophrenia. In addition, the magnitude of the activity was negatively correlated with age in this region in the controls, but not in the patients. Discussion)Our results demonstrate the availability of NIRS for evaluating the magnitude of the resting-state activity in the mPFC in schizophrenia. Further studies will be warranted to develop the current NIRS study of resting-state activity into routine psychiatry practice.		P5-10 Gamma-band auditory steady-state response is associated with plasma level of D-serine in schizophrenia 統合失調症におけるガンマオシレーションとD-セリン血漿中濃度との相関解析 Usui Kaori（臼井香）¹，越山太輔¹，切原賢治¹，多田真理子¹，永井達哉¹，藤岡真生¹，小池進介^1,2，菅心^1,3，荒木剛¹，笠井清登¹ ¹Department of Neuropsychiatry, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan ²University of Tokyo Institute for Diversity & Adaptation of Human Mind (UTIDAHM), Tokyo, Japan ³Department of Rehabilitation, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan Gamma-band auditory steady-state response (ASSR), which is considered to reflect γ-aminobutyric acid (GABA)-ergic interneuron function, is a candidate of useful biomarker for investigating new treatments in schizophrenia. The N-methyl-D-aspartate (NMDA) receptor hypofunction is one of the major hypotheses of pathophysiology in schizophrenia, and alteration of plasma levels of glutamatergic amino acids such as D-serine, co-agonists of NMDA receptors, have been reported in patients with schizophrenia. Recent animal studies have shown that NMDA receptor dysfunction in parvalbumin-positive GABAergic interneurons causes deficits in gamma oscillations. In this study, we investigated the relationships between gamma-band ASSR and plasma levels of glutamatergic amino acids to elucidate the association between gamma-band ASSR and NMDA receptor function in patients with schizophrenia. The participants included 23 patients with schizophrenia and 22 healthy controls. Correlational analyses revealed that gamma-band ASSR, which was impaired in patients with schizophrenia compared with healthy controls at the trend level, significantly correlated with the plasma levels of D-serine (r = 0.47, p = 0.024) only in schizophrenia, but not in healthy controls. These findings suggest that gamma-band ASSR may reflect NMDA receptor function in schizophrenia. Therefore, gamma-band ASSR may serve as a biomarker for development of new treatments targeting NMDA receptor hypofunction in schizophrenia.