Genomic Relationships, Novel Loci, and Pleiotropic Mechanisms across Eight Psychiatric Disorders

Genetic influences on psychiatric disorders transcend diagnostic boundaries, suggesting substantial pleiotropy of contributing loci. However, the nature and mechanisms of these pleiotropic effects remain unclear. We performed analyses of 232,964 cases and 494,162 controls from genome-wide studies of anorexia nervosa, attention-deficit/hyper-activity disorder, autism spectrum disorder, bipolar disorder, major depression, obsessive-compulsive disorder, schizophrenia, and Tourette syndrome. Genetic correlation analyses revealed a meaningful structure within the eight disorders, identifying three groups of inter-related disorders. Meta-analysis across these eight disorders detected 109 loci associated with at least two psychiatric disorders, including 23 loci with pleiotropic effects on four or more disorders and 11 loci with antagonistic effects on multiple disorders. The pleiotropic loci are located within genes that show heightened expression in the brain throughout the lifespan, beginning prenatally in the second trimester, and play prominent roles in neurodevelopmental processes. These findings have important implications for psychiatric nosology, drug development, and risk prediction.Peer reviewe

Dissecting the Shared Genetic Architecture of Suicide Attempt, Psychiatric Disorders, and Known Risk Factors

Background Suicide is a leading cause of death worldwide, and nonfatal suicide attempts, which occur far more frequently, are a major source of disability and social and economic burden. Both have substantial genetic etiology, which is partially shared and partially distinct from that of related psychiatric disorders. Methods We conducted a genome-wide association study (GWAS) of 29,782 suicide attempt (SA) cases and 519,961 controls in the International Suicide Genetics Consortium (ISGC). The GWAS of SA was conditioned on psychiatric disorders using GWAS summary statistics via multitrait-based conditional and joint analysis, to remove genetic effects on SA mediated by psychiatric disorders. We investigated the shared and divergent genetic architectures of SA, psychiatric disorders, and other known risk factors. Results Two loci reached genome-wide significance for SA: the major histocompatibility complex and an intergenic locus on chromosome 7, the latter of which remained associated with SA after conditioning on psychiatric disorders and replicated in an independent cohort from the Million Veteran Program. This locus has been implicated in risk-taking behavior, smoking, and insomnia. SA showed strong genetic correlation with psychiatric disorders, particularly major depression, and also with smoking, pain, risk-taking behavior, sleep disturbances, lower educational attainment, reproductive traits, lower socioeconomic status, and poorer general health. After conditioning on psychiatric disorders, the genetic correlations between SA and psychiatric disorders decreased, whereas those with nonpsychiatric traits remained largely unchanged. Conclusions Our results identify a risk locus that contributes more strongly to SA than other phenotypes and suggest a shared underlying biology between SA and known risk factors that is not mediated by psychiatric disorders.Peer reviewe

A genetic investigation of sex bias in the prevalence of attention-deficit/hyperactivity disorder

Background Attention-deficit/hyperactivity disorder (ADHD) shows substantial heritability and is 2-7 times more common in males than females. We examined two putative genetic mechanisms underlying this sex bias: sex-specific heterogeneity and higher burden of risk in female cases. Methods We analyzed genome-wide autosomal common variants from the Psychiatric Genomics Consortium and iPSYCH Project (20,183 cases, 35,191 controls) and Swedish populationregister data (N=77,905 cases, N=1,874,637 population controls). Results Genetic correlation analyses using two methods suggested near complete sharing of common variant effects across sexes, with rg estimates close to 1. Analyses of population data, however, indicated that females with ADHD may be at especially high risk of certain comorbid developmental conditions (i.e. autism spectrum disorder and congenital malformations), potentially indicating some clinical and etiological heterogeneity. Polygenic risk score (PRS) analysis did not support a higher burden of ADHD common risk variants in female cases (OR=1.02 [0.98-1.06], p=0.28). In contrast, epidemiological sibling analyses revealed that the siblings of females with ADHD are at higher familial risk of ADHD than siblings of affected males (OR=1.14, [95% CI: 1.11-1.18], p=1.5E-15). Conclusions Overall, this study supports a greater familial burden of risk in females with ADHD and some clinical and etiological heterogeneity, based on epidemiological analyses. However, molecular genetic analyses suggest that autosomal common variants largely do not explain the sex bias in ADHD prevalence

Defi catches on Implementation of a curriculum for resuscitation in secondary schools

Sudden cardiac arrest is still one of the most frequent causes of death. Teaching resuscitation in schools was already successfully implemented in Scandinavian countries. Following a recommendation of the conference of german stateministers of education in June 2014, additional tuition for resuscitation is to be implemented in german schools starting in seventh grade. The present study aimed to assess the level of knowledge of seventh grade students in the field of life support and to implement curricular standards for resuscitation courses in secondary schools. Using a standardized questionnaire, students in seventh grade of five schools in Cologne were interrogated about their knowledge on resuscitation and defibrillation. This assessment was taken as basis for developing a curricular teaching concept by the Cologne heart centre in cooperation with the department of biology and technical didactics at the University of Cologne. This tutorial concept was integrated in scholar plans for the first time in the school year 2014/2015. At the end of the school year the students' knowledge got reevaluated. As expected, the technical knowledge of the interviewed students is low, however confidence in their own abilities is high. Most of the interviewed persons would be willing to perform chest compressions (72,26%) and dare using an automated external defibrillator (AED, 64,38%). The exact position of defibrillator pads cannot be precisely indicated by most students (8,40%), compression point, -depth and -frequency are known by just one third of students. Already one-time performance of the live-saving lesson resulted in a clear increase of knowledge about resuscitation. The willingness to perform resuscitation measures and confidence in their own abilities are high in seventh grade students. Therefore, the recommendation of the conference of german stateministers of education in June 2014 addresses the right target group. Long-term success of the presented educational concept will be analysed and reported in a longitudinal study

Circulating MACC1 transcripts in glioblastoma patients predict prognosis and treatment response

Glioblastoma multiforme is the most aggressive primary brain tumor of adults, but lacksreliable and liquid biomarkers. We evaluated circulating plasma transcripts of metastasis-associatedin colon cancer-1 (MACC1), a prognostic biomarker for solid cancer entities, for prediction of clinicaloutcome and therapy response in glioblastomas. MACC1 transcripts were significantly higher inpatients compared to controls. Low MACC1 levels clustered together with other prognosticallyfavorable markers. It was associated with patients’ prognosis in conjunction with the isocitratedehydrogenase (IDH) mutation status: IDH1 R132H mutation and low MACC1 was most favorable(median overall survival (OS) not yet reached), IDH1 wildtype and high MACC1 was worst (medianOS 8.1 months), while IDH1 wildtype and low MACC1 was intermediate (median OS 9.1 months).No patients displayed IDH1 R132H mutation and high MACC1. Patients with low MACC1 levelsreceiving standard therapy survived longer (median OS 22.6 months) than patients with high MACC1levels (median OS 8.1 months). Patients not receiving the standard regimen showed the worstprognosis, independent of MACC1 levels (low: 6.8 months, high: 4.4 months). Addition of circulatingMACC1 transcript levels to the existing prognostic workup may improve the accuracy of outcomeprediction and help define more precise risk categories of glioblastoma patients

Expression of activating transcription factor 5 (ATF5) is increased in astrocytomas of different WHO grades and correlates with survival of glioblastoma patients

Background: ATF5 suppresses differentiation of neuroprogenitor cells and is overexpressed in glioblastoma (GBM). A reduction of its expression leads to apoptotic GBM cell death. Data on ATF5 expression in astrocytoma WHO grade II (low-grade astrocytoma [LGA]) are scarce and lacking on recurrent GBM. Patients and methods: ATF5 mRNA was extracted from frozen samples of patients’ GBM (n=79), LGA (n=40), and normal brain (NB, n=10), quantified by duplex qPCR and correlated with retrospectively collected clinical data. ATF5 protein expression was evaluated by measuring staining intensity on immunohistochemistry. Results: ATF5 mRNA was overexpressed in LGA (sevenfold, P<0.001) and GBM (tenfold, P<0.001) compared to NB, which was confirmed on protein level. Although ATF5 mRNA expression in GBM showed a considerable fluctuation range, groups of varying biological behavior, that is, local/multifocal growth or primary tumor/relapse and the tumor localization at diagnosis, were not significantly different. ATF5 mRNA correlated with the patients’ age (r=0.339, P=0.028) and inversely with Ki67-staining (r=-0.421, P=0.007). GBM patients were allocated to a low and a high ATF5 expression group by the median ATF5 overexpression compared to NB. Kaplan–Meier analysis and Cox regression indicated that ATF5 mRNA expression significantly correlated with short-term survival (t<12 months, median survival 18 vs 13 months, P=0.022, HR 2.827) and progression-free survival (PFS) (12 vs 6 months, P=0.024). This advantage vanished after 24 months (P=0.084). Conclusion: ATF5 mRNA expression could be identified as an additional, though not independent factor correlating with overall survival and PFS. Since its inhibition might lead to the selective death of glioma cells, it might serve as a potential ubiquitous therapeutic target in astrocytic tumors

Characterization of undifferentiated rESC and riPSC under feeder-dependent and feeder-free culture conditions.

<p>(A) Rat PSCs form floating or loosely attached colonies on mitotically inactivated MEFs (MEF-2iLIF). Both cell types are positive for alkaline phosphatase activity and show SSEA-1 and Oct4 expression. Scale bars: 200 μm. (B) Flow cytometry analyses of both rPSC types for Oct4 expression. (C) Representative diploid karyograms of rPSCs originating from MEF-2iLIF conditions. P indicates the passage number under MEF-2iLIF conditions. rESCs in passage 14 showed a normal female rat karyotype (42, XX). riPSCs in passage 27 presented with an aberrant male karyotype characterized by a translocation between one homolog of chromosome 3 and the X chromosome (arrows) and two marker chromosomes (mar), presumably composed of material from chromosomes 17 and 20 which are missing. (D) No significant difference in population doubling time was detected between rESC and riPSC. Mean ± SEM, n = 20–21, unpaired t-test, P = 0.897. (E) Also in feeder-free monolayer culture (Geltrex-2iLIF), undifferentiated rPSCs express pluripotency markers. Phase contrast images reveal the typical high nucleus-to-cytoplasm ratio of pluripotent stem cells. In addition, cells show alkaline phosphatase activity, and are immuno-positive for SSEA-1 and Oct4. Scale bars: 200 μm. (F) Flow cytometry analysis revealed Oct4^pos expression levels comparable to feeder-based cultures for both rPSC types. (G) Representative karyograms of rPSCs after several passages in Geltrex-2iLIF conditions. P indicates the passage number of cells originating from MEF-2iLIF conditions plus additional passages in Geltrex-2iLIF. After 16 passages, rESC still show a normal rat karyotype in the majority of metaphase plates. After 5 passages, riPSCs also showed no differences to the karyotype found under MEF-2iLIF conditions. However, after 18 passages in Geltrex-2IiLIF, the majority of cells showed a tetraploid karyotype (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0192652#pone.0192652.s001" target="_blank">S1 Fig</a>). (H) Proliferation rates show a significant difference of rESCs versus riPSCs in Geltrex-2iLIF culture. Mean ± SEM, n = 21, unpaired t-test, *P < 0.001.</p

Expression pattern of rPSCs during differentiation.

<p>Semiquantitative RT-PCR analyses showing the expression of pluripotency (Rex-1, Nanog, Oct4), mesoderm (T-Bra), CM-specific (GATA4, Nkx2.5, α-MHC, β-MHC, Mlc2v, Mlc2a, ANP) and gap junction (Cx40, Cx43, Cx45) genes in differentiating rESCs and riPSCs derived from AMW-based EB-formation and AA-2P enhanced differentiation in dynamic suspension culture. Expression of Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) served as internal control; water and reverse transcriptase minus (RT-) indicate negative controls.</p

Cardiac differentiation of rPSCs is enhanced by ascorbic acid-2-phosphate and is scalable using agarose microwells.

<p>(A) The differentiation protocol started (D0, day 0) with preparation of hanging drops using 3x10³ undifferentiated rPSCs (expanded in MEF-2iLIF conditions) per droplet in serum-based differentiation medium (DM). After 2 days (D2), resulting EBs were transferred individually to agarose coated 96-well plates with or without addition of 100 μM AA-2P. Numbers of beating EBs were quantified on day 8, 10, 12 and 14. (B) Cell culture plate with approximately 160 hanging drops. (C,D) Efficiency of cardiac differentiation of rPSC-EBs determined by the emergence of beating EBs over time with and without AA-2P supplementation. Mean ± SEM, (n = 4–10 independent experiments with 48 EBs per biological repetition). (E) Fold increase of beating EB number after of AA-2P supplementation analyzed on day 14. Mean ± SEM, (n = 5–10 independent experiments). Unpaired t-test, *P < 0.05. (F) Protocol for rPSC differentiation expanded in Geltrex-2iLIF conditions using forced aggregation in agarose microwells for two days. Subsequent differentiation was conducted in dynamic suspension culture. (G) rPSC aggregates with different starting cell numbers after 48 h on agarose microwells. Scale bar: 200 μm. (H) Efficiency of cardiac differentiation determined by the quantification of beating EBs on day 14 (left Y-axis). Mean ± SEM, n = 9–26 independent experiments, *P < 0.02. Cardiac differentiation efficiency determined by flow cytometry analysis of cardiac troponin T (cTnT)-stained EB-derived cells on day 14 (right Y-axis). n = 3–7. (I) Exemplary histograms of cTnT-flow cytometry analyses on differentiation day 14. Isotype controls are shown in gray.</p