Parents\u27 Goals: An Analysis of Therapist Reasoning

Purpose: Illustrate the use of DDDM to develop parent-identified goals for occupational therapy and to identify underlying sensory integration factors hypothesized to be impacting participation

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

Genetic Overlap Between Alzheimer’s Disease and Bipolar Disorder Implicates the MARK2 and VAC14 Genes

Background: Alzheimer's disease (AD) and bipolar disorder (BIP) are complex traits influenced by numerous common genetic variants, most of which remain to be detected. Clinical and epidemiological evidence suggest that AD and BIP are related. However, it is not established if this relation is of genetic origin. Here, we applied statistical methods based on the conditional false discovery rate (FDR) framework to detect genetic overlap between AD and BIP and utilized this overlap to increase the power to identify common genetic variants associated with either or both traits. Methods: We obtained genome wide association studies data from the International Genomics of Alzheimer's Project part 1 (17,008 AD cases and 37,154 controls) and the Psychiatric Genetic Consortium Bipolar Disorder Working Group (20,352 BIP cases and 31,358 controls). We used conditional QQ-plots to assess overlap in common genetic variants between AD and BIP. We exploited the genetic overlap to re-rank test-statistics for AD and BIP and improve detection of genetic variants using the conditional FDR framework. Results: Conditional QQ-plots demonstrated a polygenic overlap between AD and BIP. Using conditional FDR, we identified one novel genomic locus associated with AD, and nine novel loci associated with BIP. Further, we identified two novel loci jointly associated with AD and BIP implicating the MARK2 gene (lead SNP rs10792421, conjunctional FDR=0.030, same direction of effect) and the VAC14 gene (lead SNP rs11649476, conjunctional FDR=0.022, opposite direction of effect). Conclusions: We found polygenic overlap between AD and BIP and identified novel loci for each trait and two jointly associated loci. Further studies should examine if the shared loci implicating the MARK2 and VAC14 genes could explain parts of the shared and distinct features of AD and BIP

The genetics of the mood disorder spectrum:genome-wide association analyses of over 185,000 cases and 439,000 controls

Background Mood disorders (including major depressive disorder and bipolar disorder) affect 10-20% of the population. They range from brief, mild episodes to severe, incapacitating conditions that markedly impact lives. Despite their diagnostic distinction, multiple approaches have shown considerable sharing of risk factors across the mood disorders. Methods To clarify their shared molecular genetic basis, and to highlight disorder-specific associations, we meta-analysed data from the latest Psychiatric Genomics Consortium (PGC) genome-wide association studies of major depression (including data from 23andMe) and bipolar disorder, and an additional major depressive disorder cohort from UK Biobank (total: 185,285 cases, 439,741 controls; non-overlapping N = 609,424). Results Seventy-three loci reached genome-wide significance in the meta-analysis, including 15 that are novel for mood disorders. More genome-wide significant loci from the PGC analysis of major depression than bipolar disorder reached genome-wide significance. Genetic correlations revealed that type 2 bipolar disorder correlates strongly with recurrent and single episode major depressive disorder. Systems biology analyses highlight both similarities and differences between the mood disorders, particularly in the mouse brain cell-types implicated by the expression patterns of associated genes. The mood disorders also differ in their genetic correlation with educational attainment – positive in bipolar disorder but negative in major depressive disorder. Conclusions The mood disorders share several genetic associations, and can be combined effectively to increase variant discovery. However, we demonstrate several differences between these disorders. Analysing subtypes of major depressive disorder and bipolar disorder provides evidence for a genetic mood disorders spectrum

Bipolar multiplex families have an increased burden of common risk variants for psychiatric disorders.

Multiplex families with a high prevalence of a psychiatric disorder are often examined to identify rare genetic variants with large effect sizes. In the present study, we analysed whether the risk for bipolar disorder (BD) in BD multiplex families is influenced by common genetic variants. Furthermore, we investigated whether this risk is conferred mainly by BD-specific risk variants or by variants also associated with the susceptibility to schizophrenia or major depression. In total, 395 individuals from 33 Andalusian BD multiplex families (166 BD, 78 major depressive disorder, 151 unaffected) as well as 438 subjects from an independent, BD case/control cohort (161 unrelated BD, 277 unrelated controls) were analysed. Polygenic risk scores (PRS) for BD, schizophrenia (SCZ), and major depression were calculated and compared between the cohorts. Both the familial BD cases and unaffected family members had higher PRS for all three psychiatric disorders than the independent controls, with BD and SCZ being significant after correction for multiple testing, suggesting a high baseline risk for several psychiatric disorders in the families. Moreover, familial BD cases showed significantly higher BD PRS than unaffected family members and unrelated BD cases. A plausible hypothesis is that, in multiplex families with a general increase in risk for psychiatric disease, BD development is attributable to a high burden of common variants that confer a specific risk for BD. The present analyses demonstrated that common genetic risk variants for psychiatric disorders are likely to contribute to the high incidence of affective psychiatric disorders in the multiplex families. However, the PRS explained only part of the observed phenotypic variance, and rare variants might have also contributed to disease development

Study of amyloidogenic properties of human lysozyme variants with heavy chain antibody fragments as structural probes

Les fibres amyloïdes sont des agrégats de protéines hautement organisés qui sont associés à une trentaine de maladies appelées amyloses, dont les maladies d'Alzheimer et de Parkinson et la maladie de la vache folle. L'amylose systémique à lysozyme est une amylose non-neuropathique héréditaire associée à sept variants de la protéine (Y54N, I56T, F57I, W64R, D67H, F57I/T70N et W112R/T70N). Ces protéines forment des fibres amyloïdes extracellulaires qui se déposent dans de nombreux tissus et organes tels que le foie, la rate et les reins. Il a été montré que les mutations I56T et D67H diminuent la stabilité et la coopérativité globale de la protéine. Ainsi, dans des conditions proches des conditions physiologiques, ces variants forment, in vitro, transitoirement un état intermédiaire dans lequel le domaine β et l'hélice C se déplient de manière coopérative, alors que le reste du domaine α conserve sa structure native. La formation d'interactions intermoléculaires entre les régions dépliées serait à l'origine du processus d'agrégation qui conduit à la formation et au dépôt de fibres amyloïdes dans les tissus des patients porteurs de ces mutations. Il a également été montré que la liaison de trois fragments d'anticorps à chaînes lourdes de camélidés (VHH), dirigés contre le lysozyme humain de sauvage, inhibe in vitro la formation de fibres amyloïdes par les variants D67H et I56T. Ces trois VHH se lient à des régions différentes du lysozyme et inhibent la formation de fibres amyloïdes selon différents mécanismes. Au cours de ce doctorat, seize nouveaux VHH spécifiques du lysozyme humain ont été générés. Des expériences de liaisons compétitives suivies par résonnance plasmonique de surface ont montré que les 16 VHH se lient à cinq épitopes distincts à la surface du lysozyme. Quatre d’entre eux sont capables de se lier au lysozyme dans les conditions utilisées in vitro pour induire la formation de fibres amyloïdes par les variants du lysozyme. Leur site de liaison a été déterminé par RMN. Deux d'entre eux reconnaissent des épitopes différents de ceux des trois VHH caractérisés précédemment. Des expériences d’échange H/D analysés par spectrométrie de masse ont montré que les 4 VHH ont des capacités différentes à restaurer la coopérativité globale du variant D67H du lysozyme. L’analyse de l’ensemble des résultats nous a permis d’identifier qu'elles sont les régions du lysozyme qui doivent être affectées par la liaison d'un VHH afin de restaurer la coopérativité globale de la protéine. La liaison simultanée aux deux domaines (i.e. α et β) semble être le dénominateur commun de tous les VHH restaurant la coopérativité globale du variant D67H. La liaison aux hélices B et C, ainsi que de l'interface des deux domaines semble aussi contribuer à la restauration de la coopérativité globale. A l'inverse, une liaison qui perturbe la partie N-terminale ne permet pas de restaurer la coopérativité globale de la protéine. La liaison d’un VHH, cAb-HuL9a, au variant D67H du lysozyme inhibe de manière similaire la formation de l’intermédiaire partiellement déplié, l'élongation de fibres amyloïdes préformées et la formation de fibres amyloïdes in vitro. Cette observation est en accord avec l’hypothèse selon laquelle, la formation de cet intermédiaire est à l’origine de l’amyloïdogénicité des variants du lysozyme. Afin d'étudier les effets des mutations amyloïdogéniques récemment identifiées sur les propriétés du lysozyme et ainsi obtenir une meilleure connaissance du mécanisme de formation des fibres amyloïdes, il est nécessaire de produire ces variants en grande quantité. Les variants D67H, I56T et F57I sont produits dans Aspergillus niger. L'expression des variants dans ce micro-organisme est toutefois particulièrement chronophage alors que les rendements de production sont relativement faibles. Les tentatives d'utiliser d'autres systèmes de production tels que Pichia pastoris ou le système bacculovirus n'ont pas été concluantes. Aussi, dans le cadre de mon doctorat, j’ai étudié la possibilité de produire le variant D67H sous forme de corps d'inclusion dans Escherichia. coli. Un protocole permettant de produire 20 mg de protéine sous forme de corps d’inclusion par litre de culture a été développé. Différents stratégies ont été testées pour replier la protéine à partir des corps d’inclusion. Cette approche a permis d’obtenir une protéine ayant 90% d’activité spécifique du lysozyme. Le rendement massique obtenu après repliement est néanmoins très faible.Étude des propriétés des variants amyloïdogéniques du lysozyme humain à l'aide de fragments d’anticorps à chaînes lourdes comme sondes structurale

Production of four amyloidogenic variants of human lysozyme as inclusion bodies in Escherichia coli

Six variants of human lysozyme (I56T, F57I, W64R, D67H, F57I/T70N and W112R/T70N) are associated with a hereditary non-neuropathic systemic amyloidosis. This disease involved an extra cellular deposition of amyloid fibrils made of lysozyme variants in a wide range of organs such as liver, spleen and kidneys [1]. The characterisation at the molecular level of two variants, I56T and D67H, has shown that these mutations reduce the stability and more particularly the global cooperativity of the protein. Consequently, under physiologically relevant conditions, these variants can transiently populate a partially unfolded state in which the beta-domain and the C-helix are cooperatively unfolded while the rest of the protein remains native like [1]. The formation of intermolecular interactions between the regions that are unfolded in this intermediate state is likely to be a fundamental trigger of the aggregation process that ultimately leads to the formation and deposition of fibrils in tissues. In order to study the effects of the other amyloidogenic mutations on the properties of lysozyme and thus to get more insight in the mechanism of amyloid formation, it is necessary to produce them in large quantities. The D67H, I56T and F57I variants are currently produced in Aspergillus niger; the expression in this organism is, however, time consuming and the yield is very low. The attempts to use alternative systems such as Pichia pastoris [2], Saccharomyces cerevisiae, and Arabidopsis thaliana have not been conclusive so far. In this work, we have produced the four single-point lysozyme variants as inclusion bodies in Escherichia coli and explored the possibility to refold them. [1] Dumoulin & al., (2006) Acc. Chem. Res., 39, 603 - 610 [2] Kumita & al., (2006) FEBS J., 273, 711-72

Camelid single-domain antibody fragments as structural probes to study the mechanism of human lysozyme fibrils formation

Six variants of human lysozyme (single-point mutations I56T, F57I, W64R, D67H and double mutations F57I/T70N, W112R/T70N) are associated with a hereditary non-neuropathic systemic amyloidosis. These proteins form extracellular amyloid fibrils that deposit in a wide range of tissues and organs such as liver, spleen and kidneys where they cause damages [1]. It was shown that the D67H and I56T mutations cause a loss in stability and more particularly a loss of global cooperativity of protein [1]. Consequently, under physiologically relevant conditions, these variants can transiently populate a partially unfolded state in which the beta-domain and the C-helix are cooperatively unfolded while the rest of the protein remains native like [1]. The formation of intermolecular interactions between the regions that are unfolded in this intermediate state is likely to be a fundamental trigger of the aggregation process that ultimately leads to the formation and deposition of fibrils in tissues. The binding of three variable domain of camelid antibodies – also named nanobodies - (cAb-HuL 6 [2], cAb-HuL 5 and cAb-HuL 22 [3]) raised against the wild type human lysozyme inhibit in vitro the formation of amyloid fibrils by the lysozyme variants. These three nanobodies bind on different regions of lysozyme and act as amyloid fibrils inhibitor through different mechanisms. On one hand, cAb-HuL 6 and cAb-HuL 22 stabilize the native state of the lysozyme variants thus restoring the global cooperativity characteristic of the wild-type protein. On the other, cAb-HuL 5 probably acts by binding soluble prefibrillar aggregates. In the present work, sixteen other nanobodies specific of human lysozyme have been generated. Competition experiments have shown that they bind to five non overlapping epitopes. The effects of the binding of these nanobodies on the stability of the D67H variant of human lysozyme and on its aggregation into amyloid fibrils will be discussed. References [1] Dumoulin et al, (2006) Acc. Chem. Res, 39, 603-610. [2] Dumoulin et al, (2003) Nature, 424, 783-788. [3] Chan et al. (2008) Biochemistry, 47,11041-11054

Nanobodies as structural probes to investigate the mechanism of fibril formation by the amyloidogenic variants of human lysozyme.

Six variants of human lysozyme (single-point mutations I56T, F57I, W64R, D67H and double mutations F57I/T70N, W112R/T70N) are associated with a hereditary non-neuropathic systemic amyloidose. These proteins form extracellular amyloid fibrils that deposit in a wide range of tissues and organs such as liver, spleen and kidneys where they cause damages [1]. It was shown that the D67H and I56T mutations cause a loss in stability and more particularly a loss of global cooperativity of protein [1]. Consequently, under physiologically relevant conditions, these variants can transiently populate a partially unfolded state in which the beta-domain and the C-helix are cooperatively unfolded while the rest of the protein remains native like [1]. The formation of intermolecular interactions between the regions that are unfolded in this intermediate state is likely to be a fundamental trigger of the aggregation process that ultimately leads to the formation and deposition of fibrils in tissues. The binding of three variable domain of camelid antibodies – also named nanobodies - (cAb-HuL 6 [2], cAb-HuL 5 and cAb-HuL 22 [3]) raised against the wild type human lysozyme inhibit in vitro the formation of amyloid fibrils by the lysozyme variants. These three nanobodies bind on different regions of lysozyme and act as Amyloid fibrils inhibitor through different mechanisms. On one hand, cAb-HuL 6 and cAb-HuL 22 stabilize the native state of the lysozyme variants thus restoring the global cooperativity characteristic of the wild-type protein. On the other, cAb-HuL 5 probably acts by binding soluble prefibrillar aggregates. In the present work, sixteen other nanobodies specific of human lysozyme have been generated. Competition experiments have shown that they bind to five non overlapping epitopes. The effects of the binding of these nanobodies on the stability of the D67H variant of human lysozyme and on its aggregation into amyloid fibrils will be discussed