Detection of the Copy Number Variants of Genes in Patients with Familial Cardiac Diseases by Massively Parallel Sequencing.

INTRODUCTION: The implication of copy number variations in familial heart disease is known, although in-depth knowledge is lacking; hence, more studies are needed to further our understanding. Massively parallel sequencing, thanks to its recent surge in use, is emerging as a valid tool for the detection of this type of variant, through the use of appropriate software. METHODS: We conducted a study with 182 patients diagnosed with mendelian cardiovascular diseases who underwent sequencing using a cardiac gene panel and then a specific calling process for copy number variations (CNVs) with ExomeDepth software, which provides us with a Bayes factor (BF), a score of the probability that a CNV detected is true. RESULTS: After a rigorous CNV prioritization process, we confirmed the variants obtained by MLPA or SNP-based array, finding three real CNVs in five individuals in the MYH11, FBN1 and PDMI7 genes. CONCLUSION: The confirmed CNVs present in all cases BF values > 60, thus establishing a threshold to consider real CNVs in the calling process carried out by ExomeDepth on our gene panel.

Combination of late gadolinium enhancement and genotype improves prediction of prognosis in non-ischaemic dilated cardiomyopathy.

AIMS: Genotype and left ventricular scar on cardiac magnetic resonance (CMR) are increasingly recognized as risk markers for adverse outcomes in non-ischaemic dilated cardiomyopathy (DCM). We investigated the combined influence of genotype and late gadolinium enhancement (LGE) in assessing prognosis in a large cohort of patients with DCM. METHODS AND RESULTS: Outcomes of 600 patients with DCM (53.3 ± 14.1 years, 66% male) who underwent clinical CMR and genetic testing were retrospectively analysed. The primary endpoints were end-stage heart failure (ESHF) and malignant ventricular arrhythmias (MVA). During a median follow-up of 2.7 years (interquartile range 1.3-4.9), 24 (4.00%) and 48 (8.00%) patients had ESHF and MVA, respectively. In total, 242 (40.3%) patients had pathogenic/likely pathogenic variants (positive genotype) and 151 (25.2%) had LGE. In survival analysis, positive LGE was associated with MVA and ESHF (both, p < 0.001) while positive genotype was associated with ESHF (p = 0.034) but not with MVA (p = 0.102). Classification of patients according to genotype (G+/G-) and LGE presence (L+/L-) revealed progressively increasing events across L-/G-, L-/G+, L+/G- and L+/G+ groups and resulted in optimized MVA and ESHF prediction (p < 0.001 and p = 0.001, respectively). Hazard ratios for MVA and ESHF in patients with either L+ or G+ compared with those with L-/G- were 4.71 (95% confidence interval: 2.11-10.50, p < 0.001) and 7.92 (95% confidence interval: 1.86-33.78, p < 0.001), respectively. CONCLUSION: Classification of patients with DCM according to genotype and LGE improves MVA and ESHF prediction. Scar assessment with CMR and genotyping should be considered to select patients for primary prevention implantable cardioverter-defibrillator placement.

Searching for genetic modulators of the phenotypic heterogeneity in Brugada syndrome.

In Brugada syndrome, even within the same family where all affected individuals share the same mutation, phenotypic variation is prominent, with variable penetrance and expressivity, presenting different degrees of involvement. It is difficult to establish a direct correlation between genotype and phenotype to predict prognosis in complications and risk of sudden death. The factors that modulate this inter- and intra-familial phenotypic variability remain to be determined. With the intention of testing whether other genetic factors, in addition to the causal mutation in SCN5A, may have a modulating effect on the Brugada phenotype and the risk of sudden death, we have studied 8 families with a causal variant in SCN5A with at least two affected individuals, one of whom has suffered cardiac arrest or sudden death. Whole exome sequencing was performed looking for additional variants that modify the phenotype and allow us to predict a better or worse prognosis for the evolution of the disease. The results did not show any clear genetic modifier; nevertheless, highlight the possible implication of the cholesterol and fibrosis pathways, as well as the circadian rhythm, as possible modulators of Brugada syndrome phenotype.

Six-minute walk test distance and resting oxygen saturations but not functional class predict outcome in adult patients with Eisenmenger syndrome.

BACKGROUND: Eisenmenger syndrome (ES) represents the extreme manifestation of pulmonary arterial hypertension in patients with congenital heart disease, associated with significant exercise intolerance and mortality. Even though of six-minute-walk-test (6MWT) is routinely used in these patients, little is known about its prognostic value in comparison to functional class. METHODS AND RESULTS: We included 210 adult patients with ES who underwent a total of 822 6MWTs. Median walking distance (6MWD) was 330 m [IQR 260-395], oxygen saturation (SO2) at baseline 86% [IQR 82-91%] and SO2 at peak-exercise 69% [IQR 60-80%]. In patients commenced on advanced therapy for pulmonary hypertension, but not in the reminder, there was a significant improvement in walking distance (297±97 m vs. 325±87 m,P=0.0019), SO2 at rest (84.9±7.1 vs. 86.8±5.9%,P=0.003), SO2 at peak exercise (69.1±12.7 vs. 72.3±12.2%,P=0.04) and NYHA functional class (P=0.0047). During a follow up of 3.3 years, 29 patients died. On time-dependent Cox analysis, 6MWD (HR 0.94 per 10 m, 95%CI: 0.91-0.97,P<0.001) and baseline SO2 (HR 0.90, 95%CI:0.86-0.94,P<0.0001) were predictors of death. In contrast, age, functional class, peak-exercise SO2 and SO2 change were not related to mortality. A three-fold increased risk of death was identified in patients not reaching a 6MWD of 350 m or with baseline SO2 below 85%. CONCLUSIONS: The 6MWD and resting SO2, but not functional class were predictive of outcome in this contemporary cohort of Eisenmenger patients and should be incorporated in both risk stratification and management algorithms for these patients.

Disease targeting therapies in patients with Eisenmenger syndrome: response to treatment and long-term efficiency.

OBJECTIVES: To examine long-term efficacy of disease targeting therapies (DTT) in patients with Eisenmenger syndrome. METHODS: All adult patients with Eisenmenger syndrome treated with DTT at our institution were included. Functional class (FC), oxygen saturation and 6-minute walk test distance (6 MWTd) were analysed retrospectively. RESULTS: Between 2002 and 2010, 79 Eisenmenger patients (21 males, 16 with Down syndrome) aged 34 ± 10 years (range 17-68 years) were included. Median follow-up was 3.3 years (range 0.2 to 8.9 years). 6 MWTd increased early after initiation of DTT, with a plateau after approximately 3 years and no obvious trend towards a deterioration on average during longer-term follow-up. Two patients died during follow-up and escalation of treatment was required in 18 patients after a median period of 2.5 years. Escalation of therapy was also associated with an increase in 6 MWTd. In addition, FC improved on DTT and oxygen saturation, increased, both at rest and peak exercise. This effect was more pronounced in the patients with the lowest baseline oxygen saturation at rest. CONCLUSIONS: Long-term DTT is safe and improves objective exercise capacity and subjective symptoms. Response to DTT was typically observed early after initiation of DTT and was, on average, maintained longer-term. However, 1 in 5 patients required escalation of DTT, with time, due to symptomatic deterioration and this was associated with an afresh improvement in 6 MWTd.

Increased expression of fatty-acid and calcium metabolism genes in failing human heart.

BACKGROUND: Heart failure (HF) involves alterations in metabolism, but little is known about cardiomyopathy-(CM)-specific or diabetes-independent alterations in gene expression of proteins involved in fatty-acid (FA) uptake and oxidation or in calcium-(Ca(2+))-handling in the human heart. METHODS: RT-qPCR was used to quantify mRNA expression and immunoblotting to confirm protein expression in left-ventricular myocardium from patients with HF (n = 36) without diabetes mellitus of ischaemic (ICM, n = 16) or dilated (DCM, n = 20) cardiomyopathy aetiology, and non-diseased donors (CTL, n = 6). RESULTS: Significant increases in mRNA of genes regulating FA uptake (CD36) and intracellular transport (Heart-FA-Binding Protein (HFABP)) were observed in HF patients vs CTL. Significance was maintained in DCM and confirmed at protein level, but not in ICM. mRNA was higher in DCM than ICM for peroxisome-proliferator-activated-receptor-alpha (PPARA), PPAR-gamma coactivator-1-alpha (PGC1A) and CD36, and confirmed at the protein level for PPARA and CD36. Transcript and protein expression of Ca(2+)-handling genes (Two-Pore-Channel 1 (TPCN1), Two-Pore-Channel 2 (TPCN2), and Inositol 1,4,5-triphosphate Receptor type-1 (IP3R1)) increased in HF patients relative to CTL. Increases remained significant for TPCN2 in all groups but for TPCN1 only in DCM. There were correlations between FA metabolism and Ca(2+)-handling genes expression. In ICM there were six correlations, all distinct from those found in CTL. In DCM there were also six (all also different from those found in CTL): three were common to and three distinct from ICM. CONCLUSION: DCM-specific increases were found in expression of several genes that regulate FA metabolism, which might help in the design of aetiology-specific metabolic therapies in HF. Ca(2+)-handling genes TPCN1 and TPCN2 also showed increased expression in HF, while HF- and CM-specific positive correlations were found among several FA and Ca(2+)-handling genes.

La insuficiencia renal crónica se asocia a un peor pronóstico tras implante de stent liberador de sirolimus / Chronic renal failure is associated with worse outcome after implantation of sirolimus eluting stent

Fundamento y objetivos: La insuficiencia renal crónica (IRC) es una patología emergente en los países desarrollados, asociándose esta además con una elevada prevalencia de enfermedad coronaria. Nuestro objetivo es determinar la influencia de la IRC en la aparición de eventos cardiovasculares adversos tras implante de un stent liberador de sirolimus (SLS) en una población no seleccionada.Pacientes y método: Estudio retrospectivo de una cohorte de 461 pacientes a los que se les implantó, de forma consecutiva, uno o más stents liberadores de sirolimus entre septiembre de 2002 y diciembre de 2005 en nuestro centro. Evaluamos la incidencia de eventos adversos durante el seguimiento y su relación con la IRC. Utilizamos la ecuación de MDRD para el cálculo del filtrado glomerular.Resultados: El tiempo medio de seguimiento fue de 42 (DS±13) meses. La edad media fue 61,3 (DS±11,8) años, y el 85,5% eran varones. En total presentaban IRC 50 (11%) pacientes. En un modelo multivariado ajustado por edad, sexo, fracción de eyección, clase killip y la presencia de hipertensión, DM, anemia o trombosis del stent, la IRC se comportó como un potente predictor de mortalidad tanto al año como al final del seguimiento (RR=3,82; intervalo de confianza del 95%, 1,41–10,33, p=0,008) y como un factor independiente de riesgo aumentado de restenosis (RR=3,47; IC del 95%, 1,01–11,97; p=0,045) comparado con los pacientes sin IRC. Aunque con una mayor tendencia hacia el grupo de pacientes con IRC, no se observaron diferencias significativas en la incidencia de trombosis del stent (8% frente a 3,4% respectivamente, p=0,109) ni en la necesidad de nueva revascularización sobre el vaso diana (18,8% frente a 10,8% respectivamente, p=0,094).Conclusiones: La presencia de IRC en pacientes con enfermedad coronaria se asocia con un aumento en la incidencia de restenosis y es un potente predictor de mortalidad tras el implante de un stent liberador de sirolimus (AU)

Background and objectives: Chronic renal failure (CRF) is an emergent pathology in industrialized countries and is associated with high prevalence of coronary artery disease. Our aim is to determine the influence of CRF in the appearance of adverse cardiovascular events after sirolimus-eluting stent implantation in a non selected cohort.Patients and methods: Observational retrospective study with a cohort of 461 patients who received one or more sirolimus-eluting stent between September 2002 and December 2005 at our institution. We evaluated the incidence of adverse cardiovascular events during the follow-up period and their relation with chronic kidney disease. We used the abbreviated Modification of Diet in Renal Disease (MDRD) equation to calculate the GFR.Results: The mean follow-up was 42 months (SD±13) and the mean age was 61±11 years and 85 percent of the group were men. Chronic renal failure was present in 50 patients, 11 percent of the cohort. In a multivariate model, after adjustment for age, sex, left ventricle election fraction, anemia, diabetes, hypertension, Killip class and stent thrombosis, chronic renal failure was an independent predictive factor of death from any cause (hazard ratio, 3.82; 95 percent confidence interval, 1.41–10.33, p=0.008), and an significant risk factor for restenosis (hazard ratio 3.47; 95 percent confidence interval, 1.01–11.97, p=0.045). Significant differences were not found in thrombosis between patients with or without CRF (8% vs 3.4%, p=0,109), although a trend was observed in the CRF group. There no were statistical association with need for a new target vessel revascularization (TVR) after coronary intervention either (18.8% versus 10.5%, p=0.094). Conclusions: The presence of chronic renal failure in patients with coronary disease is associated with higher risk of restenosis and is a potent predictor of mortality after sirolimus-eluting stent implantation

[Chronic renal failure is associated with worse outcome after implantation of sirolimus eluting stent]. / La insuficiencia renal crónica se asocia a un peor pronóstico tras implante de stent liberador de sirolimus.

BACKGROUND AND OBJECTIVES: Chronic renal failure (CRF) is an emergent pathology in industrialized countries and is associated with high prevalence of coronary artery disease. Our aim is to determine the influence of CRF in the appearance of adverse cardiovascular events after sirolimus-eluting stent implantation in a non selected cohort. PATIENTS AND METHODS: Observational retrospective study with a cohort of 461 patients who received one or more sirolimus-eluting stent between September 2002 and December 2005 at our institution. We evaluated the incidence of adverse cardiovascular events during the follow-up period and their relation with chronic kidney disease. We used the abbreviated Modification of Diet in Renal Disease (MDRD) equation to calculate the GFR. RESULTS: The mean follow-up was 42 months (SD ± 13) and the mean age was 61 ± 11 years and 85 percent of the group were men. Chronic renal failure was present in 50 patients, 11 percent of the cohort. In a multivariate model, after adjustment for age, sex, left ventricle election fraction, anemia, diabetes, hypertension, Killip class and stent thrombosis, chronic renal failure was an independent predictive factor of death from any cause (hazard ratio, 3.82; 95 percent confidence interval, 1.41-10.33, p = 0.008), and an significant risk factor for restenosis (hazard ratio 3.47; 95 percent confidence interval, 1.01-11.97, p = 0.045). Significant differences were not found in thrombosis between patients with or without CRF (8% vs 3.4%, p = 0,109), although a trend was observed in the CRF group. There no were statistical association with need for a new target vessel revascularization (TVR) after coronary intervention either (18.8% versus 10.5%, p = 0.094). CONCLUSIONS: The presence of chronic renal failure in patients with coronary disease is associated with higher risk of restenosis and is a potent predictor of mortality after sirolimus-eluting stent implantation.