Next-generation sequencing for diagnosis of thoracic aortic aneurysms and dissections: diagnostic yield, novel mutations and genotype phenotype correlations.

BACKGROUND: Thoracic aortic aneurysms and dissections (TAAD) are silent but possibly lethal condition with up to 40 % of cases being hereditary. Genetic background is heterogeneous. Recently next-generation sequencing enabled efficient and cost-effective examination of gene panels. Aim of the study was to define the diagnostic yield of NGS in the 51 TAAD patients and to look for genotype-phenotype correlations within families of the patients with TAAD. METHODS: 51 unrelated TAAD patients were examined by either whole exome sequencing or TruSight One sequencing panel. We analyzed rare variants in 10 established thoracic aortic aneurysms-associated genes. Whenever possible, we looked for co-segregation in the families. Kaplan-Meier survival curve was constructed to compare the event-free survival depending on genotype. Aortic events were defined as acute aortic dissection or first planned aortic surgery. RESULTS AND DISCUSSION: In 21 TAAD patients we found 22 rare variants, 6 (27.3 %) of these were previously reported, and 16 (73.7 %) were novel. Based on segregation data, functional analysis and software estimations we assumed that three of novel variants were causative, nine likely causative. Remaining four were classified as of unknown significance (2) and likely benign (2). In all, 9 (17.6 %) of 51 probands had a positive result when considering variants classified as causative only and 18 (35.3 %) if likely causative were also included. Genotype-positive probands (n = 18) showed shorter mean event free survival (41 years, CI 35-46) than reference group, i.e. those (n = 29) without any plausible variant identified (51 years, CI 45-57, p = 0.0083). This effect was also found when the 'genotype-positive' group was restricted to probands with 'likely causative' variants (p = 0.0092) which further supports pathogenicity of these variants. The mean event free survival was particularly low (37 years, CI 27-47) among the probands with defects in the TGF beta signaling (p = 0.0033 vs. the reference group). CONCLUSIONS: This study broadens the spectrum of genetic background of thoracic aneurysms and dissections and supports its potential role as a prognostic factor in the patients with the disease.

In vivo and in vitro examination of the functional significances of novel lamin gene mutations in heart failure patients.

CONTEXT: Lamin A/C (LMNA) gene variations have been reported in more than one third of genotyped families with dilated cardiomyopathy (DCM). However, the relationship between LMNA mutation and the development of DCM is poorly understood. METHODS AND RESULTS: We found that end stage DCM patients carrying LMNA mutations displayed either dramatic ultrastructural changes of the cardiomyocyte nucleus (D192G) or nonspecific changes (R541S). Overexpression of the D192G lamin C dramatically increased the size of intranuclear speckles and reduced their number. This phenotype was only partially reversed by coexpression of the D192G and wild type lamin C. Moreover, the D192G mutation precludes insertion of lamin C into the nuclear envelope when co-transfected with the D192G lamin A. By contrast, the R541S phenotype was entirely reversed by coexpression of the R541S and wild type lamin C. As lamin speckle size is known to be correlated with regulation of transcription, we assessed the SUMO1 distribution pattern in the presence of mutated lamin C and showed that D192G lamin C expression totally disrupts the SUMO1 pattern. CONCLUSION: Our in vivo and in vitro results question the relationship of causality between LMNA mutations and the development of heart failure in some DCM patients and therefore, the reliability of genetic counselling. However, LMNA mutations producing speckles result not only in nuclear envelope structural damage, but may also lead to the dysregulation of cellular functions controlled by sumoylation, such as transcription, chromosome organisation, and nuclear trafficking.

Increased frequency of organ-specific cardiac antibodies in healthy relatives of patients with dilated cardiomyopathy: evidence for autoimmunity in Polish families.

BACKGROUND AND HYPOTHESIS: Autoantibodies represent markers of autoimmune involvement and are found with increased frequency in patients and their symptom-free relatives at risk compared with normal controls. Cardiac-specific autoantibodies, detected by immunofluorescence, were found in 20% of symptom-free relative of patients with dilated cardiomyopathy (DCM) from England and Italy. The role of autoimmunity may vary in DCM patients from Poland due to ethnic differences in genetic susceptibility to autoimmune disease. METHODS: We assessed the frequency of the organ-specific cardiac autoantibodies in 162 symptom-free relatives of DCM patients [85 male, mean (SD) age 27 (18) years] and 80 control subjects from Poland. Familial DCM (> 1 affected member) was present in 4 families, nonfamilial DCM in the remaining 24 pedigrees. We performed antibody screening and noninvasive cardiological assessment in the whole group. RESULTS: The frequency of cardiac-specific autoantibodies was higher among patients with documented DCM (probands and relatives) (50%) and their symptom-free relatives (38%) than in unrelated normal subjects (10%; p = 0.0001). In 24 (86%) of the pedigrees studied, autoantibodies were found in the proband and/or in at least one family member and tended to be more common in familial than in nonfamilial DCM (50 vs. 35%, p = NS). Echocardiographic indices of left ventricular size and function were similar in relatives with and without detectable antibodies. CONCLUSIONS: The presence of cardiac-specific autoantibodies in symptom-free relatives of DCM patients provides evidence for autoimmunity in the majority (86%) of our pedigrees, including both familial and nonfamilial forms of DCM.

Determinants of prognosis in nonischemic dilated cardiomyopathy.

BACKGROUND: Dilated cardiomyopathy, a heart muscle disease of unknown cause, is characterized by high mortality and is a major cause of cardiac transplantation. It has become, therefore, increasingly important to identify patients at higher risk. The aim of this study was to assess which of the data obtained at the time of diagnosis are the best predictors of survival. METHODS AND RESULTS: One hundred forty-four patients with dilated cardiomyopathy (118 men; mean age, 39 years) were assessed clinically, noninvasively, and hemodynamically. The effect of variables derived from the evaluation on outcome (death or heart transplantation) was examined. During a mean follow-up time of 4.1 years, 68 patients (47%) died and 9 (6%) underwent heart transplantation. The 1-, 2-, and 5-year transplant-free survival rate was 79, 69, and 44%, respectively. Cox multivariate regression analysis identified three variables as independent predictors of outcome: (1) pulmonary artery systolic pressure, P = .0001; (2) left ventricular ejection fraction, P = .0013; and (3) left ventricular end-diastolic dimension, P = .007. The prognostic index was constructed from regression coefficients and parameters significant in the Cox model. The minimal prognostic index in the study group was 1.4 and the maximal was 6.0 with a corresponding 1-year survival of 98 and 18%, respectively. The validity of the prognostic index was tested in the consecutive group of 81 patients, who were followed for a mean 2.3 years. The prognostic index of the poor outcome group differed significantly from that in survivors (3.7 vs 2.9, respectively, P < .01). The sensitivity and specificity of model predictions were 68 and 52%, respectively. CONCLUSIONS: The severity of pulmonary hypertension and left ventricular dysfunction provides an independent insight into the prognosis of patients with dilated cardiomyopathy. The prognostic index is useful when assessing prognosis and may be helpful in the timing of heart transplantation.

Left ventricular enlargement is common in relatives of patients with dilated cardiomyopathy.

Idiopathic dilated cardiomyopathy is characterized by dilation and impaired contractility of one or both ventricles. Long-term prognosis is poor. Early diagnosis has the potential for substantial reduction of morbidity and mortality. Recent studies, based on echocardiographic assessment of relatives of the patients have shown that familial dilated cardiomyopathy is relatively common. The authors studied 215 relatives (mean age, 27 years; 111 male) of 38 index patients with idiopathic dilated cardiomyopathy by clinical examination, electrocardiography, and two-dimensional, M-mode and Doppler echocardiography. Seven relatives (3%) from six families were shown to have dilated cardiomyopathy. Thus, 6 of the 38 index patients (16%) had familial disease. Furthermore, left ventricular enlargement either during diastole or systole was found in 66 of 174 healthy relatives (38%). This is significantly more frequent than in our normal control population of 100 unrelated subjects studied in the same way (18%; P < .0001). These 66 relatives with left ventricular enlargement belonged to 27 of the 38 examined families (71%). Dilated cardiomyopathy was found to be familial in 16% of patients. Of the relatives examined, 41% had left ventricular abnormalities. These findings provide further evidence for a genetic background of dilated cardiomyopathy. Relatives with left ventricular enlargement may have an early stage and/or latent form of the disease.

Organ-specific cardiac autoantibodies in dilated cardiomyopathy. Frequency and clinical correlates in Polish patients.

Organ- and disease-specific cardiac autoantibodies are found in a third of dilated cardiomyopathy patients from the U.K. and Italy and represent markers of autoimmune involvement. The role of autoimmunity may vary in dilated cardiomyopathy patients from different countries due to differences in genetic susceptibility to autoimmune diseases. The aim of this study was to assess the frequency of organ-specific cardiac autoantibodies detected by immunofluorescence in a consecutive series of patients with dilated cardiomyopathy and in disease and normal control subjects from Poland. The study groups included 79 patients with idiopathic (WHO criteria) dilated cardiomyopathy, 55 patients with other cardiac disease and 60 normal subjects. Cardiac antibody tests were performed by indirect immunofluorescence on human heart; skeletal muscle was used to identify cross-reacting antibodies. The frequency of organ-specific cardiac autoantibodies was higher in patients with dilated cardiomyopathy (21/79, 27%) than in controls with other cardiac disease (1/55, 2% P < 0.001) or in normal subjects (7/60, 12% P < 0.02). Conversely, cross-reactive antibodies were detected in similar proportions in patients with dilated cardiomyopathy (5/79, 6%), disease controls (7/55, 13%) and normal subjects (6/60, 10%, P = ns). The organ-specific antibody was more common in patients with dilated cardiomyopathy with insidious onset of disease (17/34, 50%) compared to those who did not exhibit this feature (4/45, 9%, P < 0.0001). Organ- and disease-specific cardiac autoantibodies were found in 27% of Polish patients with dilated cardiomyopathy at diagnosis; this is evidence for autoimmune involvement in a subset of patients from our country, as seen in a previously reported series of Western European origin. The association of antibody status with insidious onset of symptoms is in keeping with the long latency period observed in other autoimmune disorders.

Dilated cardiomyopathy in children: clinical course and prognosis.

The clinical profile of 19 patients with dilated cardiomyopathy ages 2-18 years (mean 13.4 +/- 4 years) was reviewed to detect any factors that might be predictive for their survival. Follow-up range from 5 to 105 months (mean 39 +/- 33 months). Routine treatment consisted of digitalis and diuretics: 14 patients received antiarrhythmics, 6 received vasodilators, and 12 were managed with immunosuppression. There were 12 survivors and 7 nonsurvivors: The 1-year mortality was 21.2% and the 2-year mortality 35.8%. All deaths were within first 2 years. Of the 12 patients who survived 2 years, a significant improvement was noticed in 9. In 3 patients tachycardia-induced cardiomyopathy was diagnosed, and abolition of supraventricular tachycardia was followed by improvement and regression of cardiomegaly. Endomyocardial biopsy was performed in 16 patients. Four with a histologic diagnosis of active myocarditis survived, and in 3 of them a considerable improvement was noticed. Of the 12 patients with nonspecific histologic findings, 6 died (p < 0.05). There were no significant differences between survivors and nonsurvivors for any of the following parameters: incidence of severe heart failure (NYHA class III-IV) and severe ventricular arrhythmias (Lown class III-V), relative heart volume, echocardiographic left ventricular diastolic diameter and shortening fraction, and the hemodynamic parameters of cardiac index, left ventricular ejection fraction, left ventricular end-diastolic pressure, and left ventricular end-diastolic volume index.

[Familial dilated cardiomyopathy with autosomal mode of inheritance. Case report]. / Kardiomiopatia rozstrzeniowa z dominujacym autosomalnym sposobem dziedziczenia. Opis rodziny.

Familial occurrence of dilated cardiomyopathy is estimated by 2-20%. We present a family with dilated cardiomyopathy inherited in an autosomal dominant way. We examined 9 members of the family, most of them are asymptomatic.

[Atypical forms of hypertrophic cardiomyopathy]. / Nietypowe postaci kardiomiopatii przerostowej.

UNLABELLED: Hypertrophic cardiomyopathy is characterized by a diverse clinical and morphological spectrum. In this report we describe five patients with atypical hypertrophic cardiomyopathy and significantly dilated atria. Diagnostic difficulties are stressed. All patients underwent clinical examination, noninvasive studies including 2-D echocardiogram, cardiac catheterization. In three patients endomyocardial biopsy was taken and examined by light microscopy. All but one patient were men, ranging in age from 28 years to 40 years at initial examination. In all patients disproportion between mildly elevated or, in two cases normal filling pressures and degree of dilatation of both atria was found. Two patients C.G. and D.T. had auscultatory and roentgenographic findings of mitral stenosis. Following echocardiographic and angiocardiographic studies an increase in left and right ventricular thickness associated with dilatation of both atria was found. Endomyocardial biopsy in both cases did not show endocardial thickening or infiltrative changes. In family S, two patients W.S. and Je.S had marginal left ventricular hypertrophy. The presence of left ventricular hypertrophy in patient Ja. S and finding, after family studies, hypertrophic, obstructive cardiomyopathy in patient A.S. enabled establishing the correct diagnosis. Beside varying degree of hypertrophy in family S, mild dilation of the right ventricle and incomplete right bundle branch block were found. In two patients of family S. restrictive cardiomyopathy was found, in patient Ja. S. during cardiac catheterization at initial presentation, in patient W.S. on doppler transmitral flow velocity examination at late follow-up. Paroxysmal atrial fibrillation was the first symptom in four patients, thromboembolic event in one patient. In four patients pacemaker requirement was found. During long-term follow-up (mean 4.8 years) slowly progressive heart failure associated with further dilatation of atria is observed. Mild doses of diuretics are effective in controlling congestive symptoms. CONCLUSION: in hypertrophic cardiomyopathy significantly dilated atria and clinical signs of mitral stenosis can be present. The presence of myocardial hypertrophy is not necessary to diagnose hypertrophic cardiomyopathy. Familial studies can be helpful in establishing the correct diagnosis.

[Dilated cardiomyopathy in children and adolescents. Diagnostic problems, clinical course and prognosis]. / Kardiomiopatia rozstrzeniowa u dzieci i mlodziezy. Problemy diagnostyczne, przebieg kliniczny i rokowania.

UNLABELLED: The clinical profile of 19 patients with dilated cardiomyopathy from 2 to 18 years old (mean age 13.4 +/- 4 years) was reviewed to detect any factors that might be predictive for their survival. Follow-up study ranged from 5 to 105 months (mean 39 +/- 33 months). All patients received digitalis + diuretics, 12 were managed with immunosuppression, 16 with antiarrhythmics. There were 12 survivors and 7 nonsurvivors: the 1-year mortality was 21.2%, the 2-years mortality was 35.8%. All deaths were within first 2 years. In 12 patients who survived 2 years, significant improvement was noticed in 9 cases. Endomyocardial biopsy was performed in 16 patients. Four of them with histological diagnosis of myocarditis survived and in 3 of them a considerable improvement was noticed. Half of 12 patients with nonspecific histological findings died (p less than 0.05). There was no significant difference between survivors and nonsurvivors in all following parameters: the incidence of severe heart failure (NYHA class III-IV) and severe ventricular arrhythmias (Lown III-IV), relative heart volume, echocardiographic LVDD, haemodynamic parameters--CI, LVEF, LVEDP, LVEDVI. CONCLUSIONS: Clinical, electrocardiographic, echocardiographic and haemodynamic data are nonpredictive for survival. The most dangerous period are the first two years of illness. In long term, improvement was noticed in half of patients.

[Reversible congestive cardiomyopathy due to chronic atrio-ventricular tachycardia]. / Odwracalna kardiomiopatia rozstrzeniowa wywolana przewleklym czestoskurczem przedsionkowo-komorowym.

A case of reversible congestive cardiomyopathy induced by a chronic tachycardia is presented. During esophageal stimulation (R-P' = 307 ms, H-A' = 355 ms, A' - H = 85 ms) it was found that tachycardia is probably due to an accessory pathway. The left ventricular dimensions were 6.0 cm (systole) and 7.0 cm (diastole), ejection fraction (EF) was 15%. After 5 days of therapy with amiodarone and methyldigoxin the patient's rhythm converted to sinus rhythm. After 15 months of therapy the EF increased to 47%, end-diastolic left ventricular dimension diminished to 5.1 cm.

[Late results of immunosuppressive therapy in patients with heart failure and biopsy proven myocarditis]. / Odlegle wyniki leczenia immunosupresyjnego u chorych z niewydolnoscia serca i bioptycznie rozpoznanym zapaleniem miesnia sercowego.

Late results of 6-month immunosuppressive therapy (prednisone from 1.5 mg/kg b.w. and azathioprine 2 mg/kg b.w.) simultaneously performed with a conventional treatment were analyzed in 20 patients with heart failure of unknown origin and bioptic diagnosed myocarditis. Average patients' age was 33.8 +/- 10.7 years, mean disease duration--7.8 month, mean left ventricular ejection fraction--25.9 +/- 8.9%. Follow up period was at least 24 months in all patients. 1 patient died before the end of therapy. After 6-month immunosuppressive therapy improvement was stated in 10 patients (50%)--group A, stable disease course in 3 (15%)--group B and deterioration in remaining 6 (30%)--group C. After the next 18 month conventional therapy as many as in 8 of 10 group A patients deterioration was observed, further improvement (EF increase from 17% to 43%) in 1 female patient and a stable disease course in the another female. Of 3 group B patients in 1 further improvement was observed and a stable course in 2 remaining. Of 6 group C patients 4 died, 1 underwent cardiac transplantation, 1 female patient is still alive, but does not put herself to control examinations. Early improvement after 6-month immunosuppressive treatment does not prejudge the later prognosis.

[Endomyocardial biopsy in children and adolescents]. / Biopsja endomiokardialna u dzieci i mlodziezy.

24 endomyocardial biopsies were performed in children aged 4-18 (x = 12.9) without any complications. The endomyocardial biopsy was performed in 12 patients with the heart failure of unknown origin (6 - restrictive heart disease, 4 - dilated cardiomyopathy with arrhythmias, 2-post-inflammatory dilated cardiomyopathy), in 5 patients with a clinical diagnosis of myocarditis, in 1 with the hypertrophic cardiomyopathy, in 2 cases of the dilated cardiomyopathy without heart failure and in 1 girl with the recurrent exudative pericarditis. Active myocarditis with fibrosis was stated in 2 cases of a restrictive heart disease, mild inflammatory state in 2 patients with a clinical diagnosis of the myocarditis and in 3 others with the dilated cardiomyopathy. All of 7 patients with the myocarditis underwent the immunosuppressive therapy. In the patient with endomyocardial fibrosis the result of left ventricular biopsy corresponded with angiocardiographic diagnosis. Non-specific changes in biopsies were stated in 2 children with the restrictive cardiomyopathy and in 5 with the dilated cardiomyopathy. Bioptic, morphologic lesions in patients with the dilated cardiomyopathy did not correlate with hemodynamic parameters of contractility. Biopsies were normal in 6 subjects. Endomyocardial biopsy influenced on making a decision of the therapy in 9 of 21 patients (42.8%). Diagnosis was verified in 7 patients (38.3%) basing on endomyocardial biopsy. In 18 of 24 subjects (75%) endomyocardial biopsy contributed to the interpretation of the disease pathology.