Genome-wide association and Mendelian randomisation analysis provide insights into the pathogenesis of heart failure

Heart failure (HF) is a leading cause of morbidity and mortality worldwide. A small proportion of HF cases are attributable to monogenic cardiomyopathies and existing genome-wide association studies (GWAS) have yielded only limited insights, leaving the observed heritability of HF largely unexplained. We report results from a GWAS meta-analysis of HF comprising 47,309 cases and 930,014 controls. Twelve independent variants at 11 genomic loci are associated with HF, all of which demonstrate one or more associations with coronary artery disease (CAD), atrial fibrillation, or reduced left ventricular function, suggesting shared genetic aetiology. Functional analysis of non-CAD-associated loci implicate genes involved in cardiac development (MYOZ1, SYNPO2L), protein homoeostasis (BAG3), and cellular senescence (CDKN1A). Mendelian randomisation analysis supports causal roles for several HF risk factors, and demonstrates CAD-independent effects for atrial fibrillation, body mass index, and hypertension. These findings extend our knowledge of the pathways underlying HF and may inform new therapeutic strategies

Genome-wide association and Mendelian randomisation analysis provide insights into the pathogenesis of heart failure

Abstract: Heart failure (HF) is a leading cause of morbidity and mortality worldwide. A small proportion of HF cases are attributable to monogenic cardiomyopathies and existing genome-wide association studies (GWAS) have yielded only limited insights, leaving the observed heritability of HF largely unexplained. We report results from a GWAS meta-analysis of HF comprising 47,309 cases and 930,014 controls. Twelve independent variants at 11 genomic loci are associated with HF, all of which demonstrate one or more associations with coronary artery disease (CAD), atrial fibrillation, or reduced left ventricular function, suggesting shared genetic aetiology. Functional analysis of non-CAD-associated loci implicate genes involved in cardiac development (MYOZ1, SYNPO2L), protein homoeostasis (BAG3), and cellular senescence (CDKN1A). Mendelian randomisation analysis supports causal roles for several HF risk factors, and demonstrates CAD-independent effects for atrial fibrillation, body mass index, and hypertension. These findings extend our knowledge of the pathways underlying HF and may inform new therapeutic strategies

Genome-wide association and Mendelian randomisation analysis provide insights into the pathogenesis of heart failure

Abstract: Heart failure (HF) is a leading cause of morbidity and mortality worldwide. A small proportion of HF cases are attributable to monogenic cardiomyopathies and existing genome-wide association studies (GWAS) have yielded only limited insights, leaving the observed heritability of HF largely unexplained. We report results from a GWAS meta-analysis of HF comprising 47,309 cases and 930,014 controls. Twelve independent variants at 11 genomic loci are associated with HF, all of which demonstrate one or more associations with coronary artery disease (CAD), atrial fibrillation, or reduced left ventricular function, suggesting shared genetic aetiology. Functional analysis of non-CAD-associated loci implicate genes involved in cardiac development (MYOZ1, SYNPO2L), protein homoeostasis (BAG3), and cellular senescence (CDKN1A). Mendelian randomisation analysis supports causal roles for several HF risk factors, and demonstrates CAD-independent effects for atrial fibrillation, body mass index, and hypertension. These findings extend our knowledge of the pathways underlying HF and may inform new therapeutic strategies

Erratum: Corrigendum: Sequence and comparative analysis of the chicken genome provide unique perspectives on vertebrate evolution

International Chicken Genome Sequencing Consortium. The Original Article was published on 09 December 2004. Nature432, 695–716 (2004). In Table 5 of this Article, the last four values listed in the ‘Copy number’ column were incorrect. These should be: LTR elements, 30,000; DNA transposons, 20,000; simple repeats, 140,000; and satellites, 4,000. These errors do not affect any of the conclusions in our paper. Additional information. The online version of the original article can be found at 10.1038/nature0315

Dishevelled binds the Discs large 'Hook' domain to activate GukHolder-dependent spindle positioning in Drosophila.

Communication between cortical cell polarity cues and the mitotic spindle ensures proper orientation of cell divisions within complex tissues. Defects in mitotic spindle positioning have been linked to various developmental disorders and have recently emerged as a potential contributor to tumorigenesis. Despite the importance of this process to human health, the molecular mechanisms that regulate spindle orientation are not fully understood. Moreover, it remains unclear how diverse cortical polarity complexes might cooperate to influence spindle positioning. We and others have demonstrated spindle orientation roles for Dishevelled (Dsh), a key regulator of planar cell polarity, and Discs large (Dlg), a conserved apico-basal cell polarity regulator, effects which were previously thought to operate within distinct molecular pathways. Here we identify a novel direct interaction between the Dsh-PDZ domain and the alternatively spliced "I3-insert" of the Dlg-Hook domain, thus establishing a potential convergent Dsh/Dlg pathway. Furthermore, we identify a Dlg sequence motif necessary for the Dsh interaction that shares homology to the site of Dsh binding in the Frizzled receptor. Expression of Dsh enhanced Dlg-mediated spindle positioning similar to deletion of the Hook domain. This Dsh-mediated activation was dependent on the Dlg-binding partner, GukHolder (GukH). These results suggest that Dsh binding may regulate core interdomain conformational dynamics previously described for Dlg. Together, our results identify Dlg as an effector of Dsh signaling and demonstrate a Dsh-mediated mechanism for the activation of Dlg/GukH-dependent spindle positioning. Cooperation between these two evolutionarily-conserved cell polarity pathways could have important implications to both the development and maintenance of tissue homeostasis in animals

Cell Fate Decision Making through Oriented Cell Division

The ability to dictate cell fate decisions is critical during animal development. Moreover, faithful execution of this process ensures proper tissue homeostasis throughout adulthood, whereas defects in the molecular machinery involved may contribute to disease. Evolutionarily conserved protein complexes control cell fate decisions across diverse tissues. Maintaining proper daughter cell inheritance patterns of these determinants during mitosis is therefore a fundamental step of the cell fate decision-making process. In this review, we will discuss two key aspects of this fate determinant segregation activity, cortical cell polarity and mitotic spindle orientation, and how they operate together to produce oriented cell divisions that ultimately influence daughter cell fate. Our focus will be directed at the principal underlying molecular mechanisms and the specific cell fate decisions they have been shown to control

Dsh directly binds the ‘Hook’ domain of Dlg.

<p>(<b>A</b>) Domain architectures of Dsh and Dlg are shown. Dsh (<i>top</i>) consists of an N-terminal DIX (<u>Di</u>shevelled and A<u>x</u>in) domain, a central PDZ (<i>yellow</i>; <u>P</u>ostsynaptic density-95, <u>D</u>iscs large, and <u>Z</u>O-1) domain, and a C-terminal DEP (<u>D</u>ishevelled, <u>E</u>gl-10, and <u>P</u>leckstrin) domain. Dlg (<i>bottom</i>) consists of tandem N-terminal PDZ domains followed by a C-terminal array of PDZ, SH3 (<i>blue</i>; <u>S</u>rc <u>h</u>omology-3), and GK (<i>red</i>; <u>G</u>uanylate <u>K</u>inase) domains. The sequence connecting the SH3 and GK domains has been termed the ‘Hook’ domain (<i>green</i>), which varies in length among species (∼90 amino acids in <i>Drosophila</i>) and undergoes alternative splicing, yielding the specific ‘I3-insert’-containing isoform investigated herein. (<b>B</b>) Structural representation of the SH3-Hook-GK cassette from <i>Drosophila</i> Dlg demonstrates the close association of the SH3 and GK domains. The Hook domain is mostly absent, likely due to conformational flexibility within protein crystals. The SH3-Hook domains act as repressors of GK domain-mediated protein interactions through a poorly understood allosteric mechanism. Image rendered from PDB id: 3TVT with bound ligand removed for clarity. (<b>C</b>) GST pulldown experiments demonstrate a Hook-dependent direct interaction between Dsh and Dlg. GST alone (control) or fused to the PDZ-DEP domains of Dsh (GST:Dsh) were coupled to glutathione agarose and subsequently incubated with soluble 6x-His-tagged Dlg proteins spanning the entire SH3-Hook-GK domains (Dlg) or an SH3-GK tandem with the Hook domain removed (DlgΔHook). Samples were resolved by SDS-PAGE and analyzed by Ponceau red staining (<i>top</i>; GST proteins) or α-His western blots (<i>bottom</i>; Dlg proteins). Purified Dlg proteins input to respective reactions are shown to the <i>right</i>. (<b>D</b>) Dlg binding occurs exclusively through the PDZ domain of Dsh. GST or GST:Dlg-Hook were incubated in the presence of various Dsh domains; <i>left lanes</i> – PDZ alone (indicated by *), <i>middle lanes</i> – both PDZ and DEP domains together (indicated by **), <i>right lanes</i> – DEP domain alone (indicated by ***). Dlg-Hook-specific interactions were detected for PDZ and PDZ-DEP constructs but not for the isolated DEP domain. The top image shows the Ponceau stained membrane depicting comparable input of GST constructs across all conditions; the bottom image is the α-His western blot used to detect bound His-tagged Dsh proteins (an ‘input’ lane of each is shown for reference).</p

Dsh expression activates Dlg-mediated spindle orientation through the GK interacting protein GukH.

<p>(<b>A</b>) <i>Drosophila</i> S2 cells were transfected with the SH3-Hook-GK domains of Dlg fused to the cell adhesion protein Ed (Ed:Dlg) alone or in combination with full-length myc-tagged Dsh (wild-type or the D316A mutant). In additional experiments, transfected cells were treated with RNAi directed against GukH. ‘Induced polarization’ of cortical Ed:Dlg was achieved through Ed-mediated cell contacts, and spindle orientation angles of mitotic cells were measured relative to the center of the induced Ed:Dlg crescent (<i>white dashed lines in Merge images</i>). Shown below cell images are western blots of S2 lysates (20 µg total protein loaded) transfected with either wild-type or D316A Dsh (as pMT:myc fusion constructs), which demonstrate equal protein expression (α-myc). An antibody against β-actin was used as a loading control. (<b>B</b>) Cumulative percentage plots reflect fraction of cells with spindles oriented at or below a given angle for all acquired images (n≥30 for all conditions). Ed alone (<i>black; circles</i>) and Ed fused to the entire SH3-Hook-GK (<i>orange; squares</i>) measurements populate a diagonal expected for completely random orientations between 0-90°. Deletion of the Hook domain (ΔHook; <i>blue; upward triangles</i>) or co-expression of Dsh (<i>green; downward triangles</i>) results in a leftward shift expected for cell populations with increased percent of lower angle measurements. Co-expression of the non-binding Dsh-D316A mutant (<i>red; diamonds</i>) did not improve spindle orientation. *, p <0.05 compared to Ed alone, One-way ANOVA with <i>Dunnett</i>'<i>s post-hoc</i> test. (<b>C</b>) GukH RNAi treatment blocks Dlg-mediated spindle orientation induced by Hook truncation (<i>yellow; squares</i>) or Dsh expression (<i>cyan; upward triangles</i>) to a level statistically equivalent to Ed alone (<i>black; circles</i>). (<b>D</b>) Three additional RNAi sequences were generated against distinct sequence elements of GukH and examined for confirmation of specificity. Each alternative (‘Alt #1-3’) target produced a similar loss-of-function phenotype as that in panel C. (<b>E</b>) Schematic representation of the GukH transcript and location of each RNAi target (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0114235#s2" target="_blank">Methods</a> for primer sequence information). Diverse targets include 5′ and 3′ untranslated regions along with two distinct targets within the coding region. All targets were against sequences shared universally among GukH isoforms. The average spindle orientation angle for each condition is shown in parentheses (compare to 26.3° for Ed:Dlg+Dsh in panel B).</p