FLNC Gene Splice Mutations Cause Dilated\ua0Cardiomyopathy

OBJECTIVE: To identify novel dilated cardiomyopathy (DCM) causing genes, and to elucidate the pathological mechanism leading to DCM by utilizing zebrafish as a model organism. BACKGROUND: DCM, a major cause of heart failure, is frequently familial and caused by a genetic defect. However, only 50% of DCM cases can be attributed to a known DCM gene variant, motivating the ongoing search for novel disease genes. METHODS: We performed whole exome sequencing (WES) in two multigenerational Italian families and one US family with arrhythmogenic DCM without skeletal muscle defects, in whom prior genetic testing had been unrevealing. Pathogenic variants were sought by a combination of bioinformatic filtering and cosegregation testing among affected individuals within the families. We performed function assays and generated a zebrafish morpholino knockdown model. RESULTS: A novel filamin C gene splicing variant (FLNC c.7251+1 G>A) was identified by WES in all affected family members in the two Italian families. A separate novel splicing mutation (FLNC c.5669-1delG) was identified in the US family. Western blot analysis of cardiac heart tissue from an affected individual showed decreased FLNC protein, supporting a haploinsufficiency model of pathogenesis. To further analyze this model, a morpholino knockdown of the ortholog filamin Cb in zebrafish was created which resulted in abnormal cardiac function and ultrastructure. CONCLUSIONS: Using WES, we identified two novel FLNC splicing variants as the likely cause of DCM in three families. We provided protein expression and in vivo zebrafish data supporting haploinsufficiency as the pathogenic mechanism leading to DCM

Meta-analysis of gene–environment-wide association scans accounting for education level identifies additional loci for refractive error

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/Myopia is the most common human eye disorder and it results from complex genetic and environmental causes. The rapidly increasing prevalence of myopia poses a major public health challenge. Here, the CREAM consortium performs a joint meta-analysis to test single-nucleotide polymorphism (SNP) main effects and SNP × education interaction effects on refractive error in 40,036 adults from 25 studies of European ancestry and 10,315 adults from 9 studies of Asian ancestry. In European ancestry individuals, we identify six novel loci (FAM150B-ACP1, LINC00340, FBN1, DIS3L-MAP2K1, ARID2-SNAT1 and SLC14A2) associated with refractive error. In Asian populations, three genome-wide significant loci AREG, GABRR1 and PDE10A also exhibit strong interactions with education (P<8.5 × 10(-5)), whereas the interactions are less evident in Europeans. The discovery of these loci represents an important advance in understanding how gene and environment interactions contribute to the heterogeneity of myopia

Systems genetics identifies a role for Cacna2d1 regulation in elevated intraocular pressure and glaucoma susceptibility

Glaucoma is a multi-factorial blinding disease in which genetic factors play an important role. Elevated intraocular pressure is a highly heritable risk factor for primary open angle glaucoma and currently the only target for glaucoma therapy. Our study helps to better understand underlying genetic and molecular mechanisms that regulate intraocular pressure, and identifies a new candidate gene, Cacna2d1, that modulates intraocular pressure and a promising therapeutic, pregabalin, which binds to CACNA2D1 protein and lowers intraocular pressure significantly. Because our study utilizes a genetically diverse population of mice with kno

Genome-wide study for circulating metabolites identifies 62 loci and reveals novel systemic effects of LPA

Genome-wide association studies have identified numerous loci linked with complex diseases, for which the molecular mechanisms remain largely unclear. Comprehensive molecular profiling of circulating metabolites captures highly heritable traits, which can help to uncover metabolic pathophysiology underlying established disease variants. We conduct an extended genome-wide association study of genetic influences on 123 circulating metabolic traits quantified by nuclear magnetic resonance metabolomics from up to 24,925 individuals and identify eight novel loci for amino acids, pyruvate and fatty acids. The LPA locus link with cardiovascular risk exemplifies how detailed metabolic profiling may inform underlying aetiology via extensive associations with very-low-density lipoprotein and triglyceride metabolism. Genetic fine mapping and Mendelian randomization uncover wide-spread causal effects of lipoprotein(a) on overall lipoprotein metabolism and we assess potential pleiotropic consequences of genetically elevated lipoprotein(a) on diverse morbidities via electronic health-care records. Our findings strengthen the argument for safe LPA-targeted intervention to reduce cardiovascular risk.Peer reviewe

The Consensus Molecular Subtypes of Colorectal Cancer

Users may view, print, copy, and download text and data-mine the content in such documents, for the purposes of academic research, subject always to the full Conditions of use -- https://www.nature.com/authors/policies/license.html#termsColorectal cancer (CRC) is a frequently lethal disease with heterogeneous outcomes and drug responses. To resolve inconsistencies among the reported gene expression-based CRC classifications and facilitate clinical translation, we formed an international consortium dedicated to large-scale data sharing and analytics across expert groups. We show marked interconnectivity between six independent classification systems coalescing into four consensus molecular subtypes (CMS) with distinguishing features: CMS1 (MSI Immune, 14%), hypermutated, microsatellite unstable, strong immune activation; CMS2 (Canonical, 37%), epithelial, chromosomally unstable, marked WNT and MYC signaling activation; CMS3 (Metabolic, 13%), epithelial, evident metabolic dysregulation; and CMS4 (Mesenchymal, 23%), prominent transforming growth factor β activation, stromal invasion, and angiogenesis. Samples with mixed features (13%) possibly represent a transition phenotype or intra-tumoral heterogeneity. We consider the CMS groups the most robust classification system currently available for CRC - with clear biological interpretability - and the basis for future clinical stratification and subtype-based targeted interventions

Multi-trait genome-wide association study identifies new loci associated with optic disc parameters.

Funder: All funders per study are acknowledged in the Supplementary FileA new avenue of mining published genome-wide association studies includes the joint analysis of related traits. The power of this approach depends on the genetic correlation of traits, which reflects the number of pleiotropic loci, i.e. genetic loci influencing multiple traits. Here, we applied new meta-analyses of optic nerve head (ONH) related traits implicated in primary open-angle glaucoma (POAG); intraocular pressure and central corneal thickness using Haplotype reference consortium imputations. We performed a multi-trait analysis of ONH parameters cup area, disc area and vertical cup-disc ratio. We uncover new variants; rs11158547 in PPP1R36-PLEKHG3 and rs1028727 near SERPINE3 at genome-wide significance that replicate in independent Asian cohorts imputed to 1000 Genomes. At this point, validation of these variants in POAG cohorts is hampered by the high degree of heterogeneity. Our results show that multi-trait analysis is a valid approach to identify novel pleiotropic variants for ONH

Whole-mount in situ hybridisationscreen in Zebrafish of homologues of genes involved in human diseases

0\. Titelseite/Inhaltsverzeichnis 1\. Einführung 1 1.1 Modellorganismen in der Entwicklungsbiologie 2 1.2 Zebrafisch als Modellorganismus 3 1.3 Entwicklung des Zebrafischembryos 3 1.4 Genetisch bedingte Krankheiten im Menschen und die entsprechende Zebrafischmutante 1.4.1 Blut/Haematopoiesis 5 1.4.2 Herz 8 1.4.3 Niere 9 1.4.4 Zentrales Nervensystem 10 1.5 Physiologische Methoden im Zebrafisch 12 1.6 Zielsetzung der Arbeit 12 2\. Material und Methoden 13 2.1 Identifikation von Zebrafisch Homologen zu menschlichen Krankheitsgenen: Datenbank Konstruktion und BLAST- Suche 13 2.2 Fischhaltung und Embryonengewinnung 13 2.3 Anzüchtung von Klonen der cDNA Bibliothek RZPDp609 14 2.4 Plasmid Präparation-Minipreps und EcoR1 Verdau 16 2.5 Herstellung von Antisense-RNA-Sonden mittels DIG-Markierung 17 2.6 Manuelle in situ Hybridisierung der whole-mount Embryonen 17 2.6.1 Rehydratation der Embryonen 17 2.6.2 Prähybridisierung 18 2.6.3 Hybridisierung 18 2.6.4 Posthybridisierung 19 2.7 Automatisierte whole-mount in situ Hybridisierung 20 2.8 Photographische Dokumentation 20 2.9 Lösungen und Puffer 21 3\. Ergebnisse 22 3.1 Einführung 22 3.2.1 Identifizierung von Homologen der Zebrafisch EST Bibliothek und beim Menschen in Krankheiten involvierten Genen 22 3.2.2 Charakterisierung von Zebrafisch Homologen mit menschlichen Krankheitsgenen auf der Basis von Gensequenzen 36 3.2.3 Charakterisierung von Zebrafisch Homologen mit menschlichen Krankheitsgenen auf der Basis von Krankheiten 36 3.3 Stadien der Zebrafischentwicklung 37 3.4 Whole-mount in situ Hybridisierungsscreen 3.4.1 Screening von 150 Klonen (ESTs) 37 3.4.2 Beschreibung der Expressionsmuster 42 3.4.2.1 Blut/Haematopoiese 42 3.4.2.2 Herz 43 3.4.2.3 Niere 45 3.4.2.4 Zentrales Nervensystem 46 3.4.2.5 Andere Organe 48 4\. Diskussion 51 4.1 Einführung 51 4.2 Sequenzanalyse 51 4.3 Bedeutung für die biomedizinische Forschung 52 4.4 Korrelation zwischen Expressionsmuster und Krankheit 53 4.4.1 Klon RZPDp609A0329 54 4.4.2 Klon RZPDp609P0227 55 4.4.3 Klon RZPDp609I0626 56 4.4.4 Klon RZPDp609B2017 56 4.4.5 Klon RZPDp609K1732 57 4.4.6 Klon RZPDp609L0444 57 4.4.7 Klon RZPDp609F2227 58 4.4.8 Klon RZPDp609J0811 59 5\. Zusammenfassung 60 6\. Literatur 61 7\. Abkürzungsverzeichnis 69 8\. Danksagung 71Der Zebrafisch hat sich in den letzten Jahren zu einem wichtigen Modellorganismus in der Molekulargenetik und Embryologie entwickelt, da sich in ihm vielseitige experimentelle Ansätze aus beiden Gebieten vereinigen lassen. Nach wie vor sind jedoch viele seiner Gene unbekannt. Das Zebrafisch EST Projekt der Washington University in St. Louis (USA) hat zum Ziel, neue Gene zu finden und zu charakterisieren. Diese Arbeit konzentrierte sich dabei auf diejenigen Klone aus der EST Bibliothek (Inhalt ~25 000 Klone), welche Homologien zu Genen aufweisen, die beim Menschen in Krankheitsvorgängen beschrieben sind. Eine Datenbank von 922 Genen, die beim Menschen als in Krankheiten impliziert ausgewiesen sind (Stand Dezember 1999), bildete die Grundlage dieser Arbeit. Für die Klonselektion wurde ein unteres Limit der Homologie von 70% auf Aminosäurenebene zwischen dieser Datenbank und den Zebrafisch EST-Sequenzen festgelegt. 186 Klone erfüllten dieses Kriterium. 150 von ihnen wurden mittels whole-mount-in-situ Hybridisierung (RNA-antisense Sonden) auf ihr Expressionsverhalten während der Entwicklung des Zebrafischembryos in einem Stadium von 6, 10, 14, 18, 24 und 36 Stunden nach der Befruchtung untersucht. Bei 39 differentiell exprimierten Klonen (26%) konnte man eine Aussage zu räumlicher und zeitlicher Expression machen. Acht Klone wurden beispielhaft in dieser Arbeit analysiert. Diese Klone sind bevorzugte Kandidaten für eine weitere funktionelle Analyse. Durch die Sequenzverifikation der ausgewählten Klone mit den mittels high-through-output Technologie ermittelten Sequenzen des Washington EST Projekt kann man ferner eine Aussage zur Höhe des Anteils der falschen Zuordnung von Sequenz und Klonen machen. Bei den in dieser Arbeit untersuchten 186 Klonen waren 46 Sequenzen (24%) nicht korrekt. Dieser Anteil bewegt sich knapp über der Annahme der Washington University für falsche Zuordnungen (15-20%).The zebrafish has emerged in recent years as a valuable organism in molecular biology and embryology since versatile experimental procedures from both areas can be combined in it. However many of its genes are unknown. The Zebrafisch EST project of the Washington University in St. Louis (USA) has the goal to find and characterize new genes. This work concentrated on those clones from an EST library (content ~25 000 clones) which are homologues to genes described as involved in human diseases. A database of 922 genes, which are proved to be implied in human diseases (as at December 1999), formed the basis of this work. For the clone selection the limit of the homology was specified to 70% on amino acid level between this database and the zebrafish EST sequences. 186 clones fulfilled this criterion. 150 of them were examined by means of whole mount in situ hybridisation (RNA antisense probes) for their expression pattern during the development of the zebrafish embryos in a stage by 6, 10, 14, 18, 24 and 36 hours after fertilization. For 39 clones (26%) one could make a statement concerning the spatial and temporal expression. Eight clones were analyzed as specimens in this work. These clones are preferential candidates for a further functional analysis. By the sequence verification of the selected clones with the sequences obtained from the Washington EST project, determined by means of high through output technology one can make a statement concerning the height of the portion of the wrong allocation of sequence and clones. Within this work 46 sequences of 186 clones were incorrect (24%). This portion moves scarcely over the percentage submitted from the Washington University for wrong allocations (15-20%)

Prediction of Chronic Kidney Disease (CKD) Progression in Children by Urinary Neutrophil-Associated Lipocalin (NGAL)

WOS: 000321387201044

Prediction of Chronic Kidney Disease (CKD) Progression in Children by Urinary Neutrophil-Associated Lipocalin (NGAL)

WOS: 00032138720104