Atlas of the clinical genetics of human dilated cardiomyopathy

AIM: Numerous genes are known to cause dilated cardiomyopathy (DCM). However, until now technological limitations have hindered elucidation of the contribution of all clinically relevant disease genes to DCM phenotypes in larger cohorts. We now utilized next-generation sequencing to overcome these limitations and screened all DCM disease genes in a large cohort. METHODS AND RESULTS: In this multi-centre, multi-national study, we have enrolled 639 patients with sporadic or familial DCM. To all samples, we applied a standardized protocol for ultra-high coverage next-generation sequencing of 84 genes, leading to 99.1% coverage of the target region with at least 50-fold and a mean read depth of 2415. In this well characterized cohort, we find the highest number of known cardiomyopathy mutations in plakophilin-2, myosin-binding protein C-3, and desmoplakin. When we include yet unknown but predicted disease variants, we find titin, plakophilin-2, myosin-binding protein-C 3, desmoplakin, ryanodine receptor 2, desmocollin-2, desmoglein-2, and SCN5A variants among the most commonly mutated genes. The overlap between DCM, hypertrophic cardiomyopathy (HCM), and channelopathy causing mutations is considerably high. Of note, we find that >38% of patients have compound or combined mutations and 12.8% have three or even more mutations. When comparing patients recruited in the eight participating European countries we find remarkably little differences in mutation frequencies and affected genes. CONCLUSIONS: This is to our knowledge, the first study that comprehensively investigated the genetics of DCM in a large-scale cohort and across a broad gene panel of the known DCM genes. Our results underline the high analytical quality and feasibility of Next-Generation Sequencing in clinical genetic diagnostics and provide a sound database of the genetic causes of DCM

Pregled znanstvenih napredaka u učinskoj elektronici usmjerenih ka osiguravanju efikasnog rada i dužeg životnog vijeka PEMgorivih ćelija

This article focuses on the main issues that affect the lifetime and performance of proton-exchange membrane fuel cells. The short lifespans of these fuel cells represent a barrier to their massive commercialization and usage in mobile and stationary applications. As fuel cell is a very complex system, a lot of knowledge of different areas is required, such as chemistry, electricity and mechanics, in order to completely understand its operation and all the problems that can occur during it. It is for this reason that an interdisciplinary approach needs to be taken when designing fuel-cell energy systems. This paper focuses on identifying and solving those issues that negatively affect the lifetime and performance of fuel cells. It is hoped that this article would be a valuable aid for power electronics’ researchers and engineers for better understanding the presented issues and a useful guide for solving them with the use of proper power electronic-devices. Initially, the basic operation and structure of a proton-exchange membrane fuel cell is explained. Three main issues that can occur during operation of a mobile or stationary fuel cell energy system are pointed out and discussed in details, on the basis of the state-of-the-art on fuel cell technology. These issues are poor water management, reactant gas starvation and fuel cell current ripple. This article provides answers as to why they occur, how they affect the fuel cell, how they can be mitigated, and what are the future trends within this research field.Članak se osvrće na ključna pitanja koja utječu na vrijeme rada i performanse gorivih ćelija s polimernom membranom kao elektrolitom. Kratak životni vijek gorivih ćelija takve vrste prepreka je njihovoj komercijalizaciji i masovnoj upotrebi u mobilnim i stacionarnim stanicama. Budući da su gorive ćelije komplicirani sustavi potrebno je znanje iz raznih područja kemije, elektrotehnike i mehanike da bi se u potpunosti mogao razumjeti njihov način rada i problemi koji se događaju. Upravo je zbog toga multidisciplinarni pristup nužnost pri razvoju sustava koji koriste gorive ćelije. Ovaj je članak usmjeren prema identifikaciji i rješavanju onih problema koji negativno utječu na životni vijek i performanse gorivih ćelija. Autori se nadaju da će se članak pokazati kao korisna pomoć i vodič istraživačima i inženjerima u domeni učinske elektronike pri susretu s navedenim problemima. Objašnjen je način rada i struktura gorive ćelije s polimernom membranom kao elektrolitom. Izložena su, i diskutirana do u detalje, tri glavna problema sa stajališta trenutačnih spoznaja u području učinske elektronike. Ti problemi su: loše upravljanje vodom, nestanak reaktantnog plina i strujni trzaji u gorivim ćelijama. Objašnjeno je zašto se ovi problemi događaju, kako utječu na gorivu ćeliju, kako ih se može spriječiti i koje su buduće perspektive istraživanja

Formin Homology 2 Domain Containing 3 (FHOD3) Is a Genetic Basis for Hypertrophic Cardiomyopathy

BACKGROUND: The genetic cause of hypertrophic cardiomyopathy remains unexplained in a substantial proportion of cases. Formin homology 2 domain containing 3 (FHOD3) may have a role in the pathogenesis of cardiac hypertrophy but has not been implicated in hypertrophic cardiomyopathy. OBJECTIVES: This study sought to investigate the relation between FHOD3 mutations and the development of hypertrophic cardiomyopathy. METHODS: FHOD3 was sequenced by massive parallel sequencing in 3,189 hypertrophic cardiomyopathy unrelated probands and 2,777 patients with no evidence of cardiomyopathy (disease control subjects). The authors evaluated protein-altering candidate variants in FHOD3 for cosegregation, clinical characteristics, and outcomes. RESULTS: The authors identified 94 candidate variants in 132 probands. The variants' frequencies were significantly higher in patients with hypertrophic cardiomyopathy (74 of 3,189 [2.32%]) than in disease control subjects (18 of 2,777 [0.65%]; p < 0.001) or in the gnomAD database (1,049 of 138,606 [0.76%]; p < 0.001). FHOD3 mutations cosegregated with hypertrophic cardiomyopathy in 17 families, with a combined logarithm of the odds score of 7.92, indicative of very strong segregation. One-half of the disease-causing variants were clustered in a small conserved coiled-coil domain (amino acids 622 to 655); odds ratio for hypertrophic cardiomyopathy was 21.8 versus disease control subjects (95% confidence interval: 1.3 to 37.9; p < 0.001) and 14.1 against gnomAD (95% confidence interval: 6.9 to 28.7; p < 0.001). Hypertrophic cardiomyopathy patients carrying (likely) pathogenic mutations in FHOD3 (n = 70) were diagnosed after age 30 years (mean 46.1 ± 18.7 years), and two-thirds (66%) were males. Of the patients, 82% had asymmetric septal hypertrophy (mean 18.8 ± 5 mm); left ventricular ejection fraction <50% was present in 14% and hypertrabeculation in 16%. Events were rare before age 30 years, with an annual cardiovascular death incidence of 1% during follow-up. CONCLUSIONS: FHOD3 is a novel disease gene in hypertrophic cardiomyopathy, accounting for approximately 1% to 2% of cases. The phenotype and the rate of cardiovascular events are similar to those reported in unselected cohorts. The FHOD3 gene should be routinely included in hypertrophic cardiomyopathy genetic testing panels

Novel Filamin C missense mutation associated with severe restrictive cardiomyopathy overlapping with left ventricular non-compaction

Abstract Introduction Filamin C gene (FLNC) missense mutations have been previously reported in association with restrictive cardiomyopathy (RCM). The association of FLNC missense variants with non-compaction cardiomyopathy has been reported only in a single proband, but familiar or functional evidence on its causative effect is limited. Overlapping traits among cardiomyopathies related to the same genetic substrate is an emerging and a challenging scenario nowadays. Purpose To report a new pathogenic FLNC missense variant in association with a particular form of restrictive/non-compaction overlapping cardiomyopathy. Methods The probands fulfill the diagnostic criteria for RCM based on current guidelines. Genetic testing in the probands was performed by NGS, using a broad gene panel (containing over 240 genes). Clinical and genetic cascade screening were expanded to first-degree relatives when it was possible. All mutation carriers underwent clinical assessment including physical examination, 12-lead ECG, echocardiography, cardiac magnetic resonance (MRI), 24h Holter monitoring, and ergometry. None of them had major systemic illnesses nor clinical symptoms of muscular involvement. Results The p.Gly2011Arg variant in the FLNC gene was the only relevant variant in the three probands. This variant is virtually absent in the general population (gnomAD). The variant showed a de novo presentation in the first family and segregated with the phenotype in the four studied relatives from the second family (three affected carriers and one unaffected non-carrier). In the third case no familial information was available. This variant affects an ultraconserved residue and is located in a relevant sub-region of Filamin-C, which is necessary for its interaction with other Z-disc proteins. Detailed clinical information is available on all carriers (n=5, 1 male). The average age at diagnosis is 17 years [1–36]. An abnormal ECG was the earliest clinical manifestation (left ventricular hypertrophy by voltage criteria and extensive repolarization abnormalities). Significant hypertrabeculations, mainly at the anterolateral wall and basal anteroseptal segments, was present in all affected carriers. Interestingly, none of them showed an abnormal late-gadolinium enhancement pattern on MRI. The four carriers who were older than 35 years were found to have severe restrictive pattern on echocardiography (functional parameters and secondary features such as bi-atrial dilation), all four suffered from limiting dyspnea, and two are under pre-transplant workup (A-II-1 and B-III-1). One of them had a cardioembolic event (femoral acute ischemia, A-II-1). One relative has recently died from advanced heart failure (B-II-2). Conclusion This is the first description on this overlapping (restrictive/non-compaction cardiomyopathy) and aggressive phenotype associated with a missense FLNC variant. This description widens the clinical spectrum related to FLNC missense mutations. Pedigrees and clinical characterization Funding Acknowledgement Type of funding source: Private company. Main funding source(s): Health in Code </jats:sec

Genetic variants as determinants of outcome in lamin A/C-related cardiac disease

Abstract Background Current guidelines for the diagnosis and management of familial dilated cardiomyopathy highlight the variables “male sex” and “non-missense type variants” as risk factors for malignant ventricular arrhythmias. Objective Quantitative evaluation of prognostic differences between different LMNA variants associated with cardio-laminopathy. Method Analysis of cardiac event-free survival (sudden death, major arrhythmic events, death from heart failure and transplantation) with Kaplan-Meier approach in relation to gender &amp; variant LMNA type. The data come from a specific database containing information on more than 1200 carriers of disease-causing LMNA variants. In the first analysis, the groups of comparison were truncating-type variants (LMNAtv) VS the global of pathogenic missense variants in the gene associated with cardiolaminopathy (LMNAm), segregated by gender. In the second analysis, it was considered missense LMNA affecting different residues (p.Arg190, p.Arg377 and p.Arg541), located in different functional domains, with enough data for comparison and with statistically different clinical behavior from that of global pathogenic variants in the gene. They were compared with the group of LMNAtv variants, as reference. The variants included were p.Arg377Cys/His, p.Arg541Cys/Ser/Gly/Pro/His and p.Arg190Trp/Gln/Pro, all of them pathogenic and associated with cardio-laminopathy. Results No significant differences were observed in survival between LMNAtv versus LMNAm variants (log rank=0.56) with slightly worse outcomes in males (log rank 0.03). Median survival time was 56 years for men compared to 60 years for women with LMNAtv, and 55 years compared to 66 years, respectively, among carriers of LMNAm (analysis A). In analysis B, statistically significant differences were observed between the groups considered (Log Rank p&amp;lt;0.001). These differences were also clinically relevant (median survival time in groups p.Arg377, LMNAtv, p.Arg190 and p.Arg541 was 60, 58, 50 and 35 years, respectively). Importantly, more than 70% of the cardiac events observed were related to major ventricular arrhythmic episodes. Conclusions This quantitative analysis demonstrates that certain missense variants in LMNA may have a similar and even more adverse clinical course than the set of truncation-type variants. These findings highlight the relevance of the specific variant rather than the variant type in guiding actionable therapies to prevent adverse outcomes. Regarding the differences observed between genders, even though they are statistically significant, their magnitude could be clinically not relevant. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Health in Code </jats:sec

Electronic circular dichroism and nuclear magnetic resonance studies of peptides derived from the FKBP52‐interacting β‐turn of the hERα ligand‐binding domain

International audienc

Marfan syndrome: genetic variant determinants of cardiovascular outcomes

Abstract Background Marfan syndrome is a systemic connective tissue disorder caused by genetic variants in the fibrillin-1 (FBN1) gene. Cardiovascular complications are the leading cause of mortality. Purpose To compare cardiovascular outcome by gender and by type of the genetic variant in the FBN1 gene. Materials and methods We analyzed clinical data on 1956 carriers and affected relatives with 1430 pathogenic or likely pathogenic genetic variants in the FBN1 gene (including cases identified in our laboratory and cases described in the literature) in whom age at last follow-up was available. We excluded patients with pathogenic or likely pathogenic genetic variants located in the so-called “neonatal region” (exon 24–32); they are recognized to have an early onset/severe phenotype. Kaplan-Meier survival curves were generated to examine gender and type of genetic variant in relation to survival free of surgical intervention or cardiovascular (CV) death (composite of deaths related to aortic dissection, heart failure/transplant, sudden, stroke or CV surgery). Genetic variants were classified as truncating (nonsense, frameshift and splicing), “missense non-neonatal” and only those missense eliminating a Cysteine residue in the non-neonatal region (“Cys non-neonatal”). Results Data were examined on 896 patients with truncating variants (53% male; 47% female) and 1060 with missense variants, located outside the “neonatal region” (54% male; 46% female). Of these, 475 were missense variants substituting a cysteine residue (52% male; 48% female). Those with truncating variants had worse prognosis versus those with missense and Cys variants (p=0.000108 and p=0.000115), with earlier onset of cardiovascular events. Overall, patients with missense variants had similar prognosis to those with missense variants eliminating a Cysteine residue. By age 65, however, almost 50% of patients with any type of variant had suffered a CV event, and with each variant type males had worse prognosis (see Figure 1). This was most evident in males aged 30 to 50 with missense variants that substituted a Cysteine residue, while female carriers of these variants had a prognosis similar to other missense variants (see Figure 2). During the first decade carriers of missense and truncating variants mainly died of heart failure. From age 10 to 50, aortic dissection was the most common event, while later other events became more frequent, e.g. vascular intervention and sudden death. Conclusion In our cohort, male carriers of pathogenic or likely pathogenic variants had worse prognosis versus females. Carriers of truncating variants had the worst CV outcomes. However, it is noteworthy that by age 65, regardless of gender or mutation type, close to 50% of patients had experienced a major CV event/death. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Health in Code Figure 2. Type by gender </jats:sec

Narrowing of the neonatal region in the FBN1 gene

Abstract Background Neonatal Marfan syndrome (MFS) is considered the most severe form of MFS and is characterized by early childhood death due to congestive heart failure. It has been suggested that genetic variants associated with this clinical presentation, cluster in a specific region between exons 24 and 32. It has been reported that patients carrying genetic variants in these exons have worse prognosis. Purpose Our purpose was to analyze cardiovascular outcome by location of the genetic variants in the “neonatal region” of the FBN1 gene. Materials and methods We analyzed clinical data on 1353 carriers and affected relatives with 683 missense pathogenic or likely pathogenic genetic variants of FBN1 gene (including cases identified in our laboratory and cases described in the literature) in whom age at last follow-up was available. Kaplan-Meier survival curves were generated to examine location of the genetic variant in the FBN1 gene in relation to survival free of surgical intervention or cardiovascular (CV) death (composite of deaths related to aortic dissection, heart failure/transplant, sudden, stroke or CV surgery). Missense genetic variants were classified as located in the “neonatal region” (residues 952–1363, corresponding to exons 24–32) and in the “non-neonatal region” (residues 45–951 and 1364–2731). In the “neonatal region”, we have also analyzed a sub-region of “over-representation” of heart failure deaths in the first year of life, which we called “critical neonatal region” (amino acids 1028–1088, corresponding to exon 25 and few residues from exon 26) in comparison to the “non-critical neonatal region”. Results Data were examined on 1060 patients with missense variants located in the “non neonatal region” and on 293 patients with missense variants located in the “neonatal region”. Of these, 96 patients were carriers of missense variants in the “critical neonatal region”, the rest of patients carried variants in the neonatal region, outside this particular domain (“non-critical neonatal region”). Patients carrying missense variants in the neonatal region had worse prognosis than those carrying variants outside this region. This poorer outcome was due to events occurring in patients carrying variants in the “critical neonatal region” (see Figure). These patients had the worse prognosis (p=0.000108, vs. the other groups). Furthermore, events in the non-critical neonatal region were similar to other missense variants located outside the neonatal region. There were no differences in the “neonatal region” when analyzing by gender. Conclusion In our cohort, the worse prognosis seen in patients carrying missense pathogenic or likely pathogenic variants in the “neonatal region” compared to the “non-neonatal region”, was due to events in patients carrying missense variants in a small subregion which we called the “critical-neonatal region” (exons 25 and 26). These patients had the worse prognosis, irrespective of gender. Funding Acknowledgement Type of funding sources: Private company. Main funding source(s): Health in Code Figure 1. Neonatal region FBN1 </jats:sec