Deregulated microRNA and mRNA expression profiles in the peripheral blood of patients with Marfan syndrome.

BACKGROUND: MicroRNAs (miRNAs) are small RNAs regulating gene expression post-transcriptionally. While acquired changes of miRNA and mRNA profiles in cancer have been extensively studied, little is known about expression changes of circulating miRNAs and messenger RNAs (mRNA) in monogenic constitutional anomalies affecting several organ systems, like Marfan syndrome (MFS). We performed integrated miRNA and mRNA expression profiling in blood samples of Marfan patients in order to investigate deregulated miRNA and mRNA networks in these patients which could serve as potential diagnostic and prognostic tools for MFS therapy. METHODS: MiRNA and mRNA expression profiles were determined in blood samples from MFS patients (n = 7) and from healthy volunteer controls (n = 7) by microarray analysis. Enrichment analyses of altered mRNA expression were identified using bioinformatic tools. RESULTS: A total of 28 miRNAs and 32 mRNAs were found to be significantly altered in MFS patients compared to controls (> 2.0-fold change, adjusted P < 0.05). The expression of 11 miRNA and 6 mRNA candidates was validated by RT-qPCR in an independent cohort of 26 MFS patients and 26 matched HV controls. Significant inverse correlations were evident between 8 miRNAs and 5 mRNAs involved in vascular pathology, inflammation and telomerase regulation. Significant positive correlations were present for 7 miRNAs with age, for 2 miRNAs with the MFS aortic root status (Z-score) and for 7 miRNAs with left ventricular end-diastolic diameter in MFS patients. In addition, miR-331-3p was significantly up-regulated in MFS patients without mitral valve prolapse (MVP) as compared with patients with MVP. CONCLUSIONS: Our data show deregulated gene and miRNA expression profiles in the peripheral blood of MFS patients, demonstrating several candidates for prognostic biomarkers for cardiovascular manifestations in MFS as well as targets for novel therapeutic approaches. A deregulation of miRNA expression seems to play an important role in MFS, highlighting the plethora of effects on post-transcriptional regulation of miRNAs and mRNAs initiated by constitutional mutations in single genes. Trial registration Nr: EA2/131/10 . Registered 28 December, 2010.

Characterization of micro-RNA Profile in the Blood of Patients with Marfan's Syndrome.

BACKGROUND: Marfan's syndrome (MFS) is an autosomal dominant inheritance disorder with a 1/5,000 live-birth prevalence. It is characterized by a wide range of clinical manifestations with more than 3,000 mutations identified in the FBN1 gene. In this study, we aimed to determine if specific patterns of circulating micro-RNAs (miRNAs) are associated with MFS-associated with cardiovascular diseases. METHODS: Microarray-based miRNA profiling was performed on blood samples of 12 MFS patients, and 12 healthy volunteers (HVs) controls and the differences in miRNA abundance between the two groups were validated using independent cohorts of 22 MFS and of 22 HV controls by real-time quantitative polymerase chain reaction (RT-qPCR). Enrichment analyses of altered miRNA abundance were predicted using bioinformatics tools. RESULTS: Altered miRNA abundance levels were determined between MFS (n = 34) and HVs (n = 34). In a screening phase, we analyzed 12 patients with MFS and 12 HVs by miRNA microarray. We found 198 miRNAs that were significantly altered in MFS patients as compared with HVs, including 16 miRNAs with a more than 1.5-fold change. Out of these 16 miRNAs, 10 showed a decreased abundance and 6 showed an increased abundance. In the validation phase, we analyzed independent cohorts of 22 MFS and of 22 HV controls by RT-qPCR. We confirmed the direction of abundance changes and the significance of different abundances between MFS patients and HVs for four miRNAs, namely, miR-362-5p, miR-339-3p, miR-340-5p, and miR-210-3p. Only the miR-150-5p showed a significant correlation with mitral valve prolapse (p = 0.010). The predicted targets for the validated miRNAs were associated with signal transduction, tissue remodeling, and cellular interaction pathways. CONCLUSION: The altered abundance level of different miRNAs in whole blood of MFS patients lays the ground to the development of novel diagnostic approaches with altered miRNAs levels associated with MFS with manifestations associated with cardiovascular diseases.

Analysis of circulating microRNAs in patients with repaired Tetralogy of Fallot with and without heart failure.

BACKGROUND: MicroRNAs (miRNAs) are a class of regulatory RNAs that regulate gene expression post-transcriptionally. Little, however, is known on the expression profile of circulating miRNAs in Tetralogy of Fallot (TOF) patients late after surgical repair. In this study, we aimed to identify the specific patterns of circulating miRNAs in blood of patients with repaired, non-syndromic TOF and to assess whether these specific miRNAs may be useful to differentiate patients with and without heart failure. METHODS: SurePrint™ 8 × 60 K Human v16 miRNA arrays were used to determine miRNA expression profiles in 15 healthy controls and 37 patients after TOF repair of whom 3 had symptomatic right heart failure. The expression levels of selected miRNAs have been validated by quantitative reverse transcription polymerase chain reaction (RT-qPCR). Enrichment analyses of altered miRNA expression were predicted using bioinformatic tools. RESULTS: Compared with healthy controls, a total of 49, 58 and 77 miRNAs were found to be significantly altered in TOF patients (TOF-all), TOF patients with (TOF-HF) and without symptomatic right heart failure (TOF-noHF) (>2.0-fold change, adjusted P < 0.05), respectively. Three miRNAs namely miR-181d-5p, miR-206 and miR-625-5p were validated by RT-qPCR in all TOF groups. The area under the receiver operating characteristic curve (AUC) for miR-181d-5p, miR-206 and miR-625-5p were 0.987, 0.993 and 0.769 in TOF-all and 0.990, 0.994 and 0.749 in TOF-noHF, respectively. Moreover, expression levels of miR-625-5p, miR-1233-3p and miR-421 were lower in TOF-HF compared to TOF-noHF (P = 0.012). CONCLUSIONS: Altered expression levels of circulating miRNAs were found in TOF patients late after surgical repair and are different to those seen in the right ventricular myocardium of infants with TOF. Expression levels of miR-421, miR-1233-3p and miR-625-5p are lower in TOF patients with symptomatic right heart failure and thus may indicate disease progression in these patients.

The novel two-dimensional strain reflects improvement and remodeling of left-ventricular function better than conventional echocardiographic parameters after aortic valve repair in pediatric patients.

We aimed to evaluate the outcome and regional and global left-ventricular (LV) function after aortic valve repair in children with congenital aortic valve disease. Thirty-two consecutive patients with a mean age of 12.62 years (4 months to 18 years) undergoing aortic valve repair due to valve stenosis (AS group, n = 21) or aortic regurgitation (AR group, n = 11) were studied during a follow-up period of 12 months regarding change and adaptation of myocardial function using conventional and novel echocardiographic methods, including two-dimensional (2D) strain echocardiogram. Conventional and 2D strain echocardiographic studies were performed and analyzed off-line using commercially available software (EchoPac 6.1.0, GE). Peak aortic valve gradient decreased from 62.04 ± 30.34 mmHg before surgery to 22.80 ± 14.13 mmHg 2 weeks after surgery and to 35.73 ± 22.11 mmHg 12 months after surgery (p = 0.01). The degree of AR decreased significantly to grade 0 in 20 children and to grade I in 12. There was a significant decrease of thickness of the interventricular septum (IVS) and posterior wall resulting in improvement of LV mass index (p = 0.007, p = 0.043, and p = 0.001, respectively). Significant decrease of myocardial thickness was found, especially in the IVS, in the AS group (p = 0.008), and a significant decrease in LV end-diastolic dimension (EDD) was found in the AR group (p = 0.007). 2D strain analysis showed that global peak strain, global systolic strain rate, and global early diastolic strain rates improved significantly for all patients during the study period after aortic valve repair (p < 0.001, p = 0.037, and p = 0.018, respectively). The global strain and strain rates correlated significantly to IVS thickness (r = 0.002 and r = 0.003, respectively), LV mass index (r = 0.02 and r = 0.015, respectively), and EDD (r = 0.26 and r = 0.005, respectively). Aortic valve repair surgery in pediatric patients results in improvement of global and regional systolic and diastolic LV parameters, which was better shown by 2D strain parameters rather than conventional echocardiographic parameters.

Concentrations of Soluble Angiotensin Converting Enzyme 2 (sACE2) in Children and Adults with and without COVID-19.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), causing the coronavirus disease 2019 (COVID-19) pandemic, leads to illness and death. Various risk factors for a severe course, such as higher age, male gender and pre-existing illnesses are known. However, pathophysiological risk factors are largely unclear. Notably, the mild course of disease in children is conspicuous. Angiotensin converting enzyme 2 (ACE2) serves as a receptor for SARS-CoV-2 and is a key enzyme in infection. Differences in the distribution of ACE2 can provide insights into different courses of COVID-19. Our aim was to elucidate the role of ACE2 as a pathophysiological risk factor by measuring soluble ACE2 (sACE2) via ELISA in blood samples (lithium-heparin-plasma or serum) of 367 individuals including children and adults with and without COVID-19. sACE2-levels were compared between the groups according to age and sex. In adults and children with COVID-19, sACE2-concentrations are significantly higher compared to healthy individuals. sACE2-levels increase with age and are lower in children compared to adults with COVID-19. Sex doesn't significantly influence sACE2-concentration. It remains unclear whether sACE2 concentrations increase because of the infection and what factors could influence this response. In conclusion, the increase of sACE2-concentration with age could indicate that ACE2 concentrations mirror increased COVID-19 severity.

Root remodeling and aortic valve repair for unicuspid aortic valve.

BACKGROUND: Unicuspid aortic valve (UAV) anatomy is occasionally encountered in adolescents or young adults and not infrequently associated with aneurysm of the ascending aorta and aortic root. To manage both defects without aortic valve replacement we propose a combination of remodeling of the aortic root combined with bicuspidization of the UAV. METHODS: Between 1 December 2007 and November 2013, 25 patients (23 males; mean age, 38 ± 12 years; range, 21 to 65 years) with aortic regurgitation as a result of UAV and aortic root dilatation underwent remodeling of the aortic root and bicuspidization of the UAV. The dilated aortic root tissue was resected, leaving the wall adjacent to the normal commissure and at 180 degrees orientation and similar height for the new commissure. The graft was configured to create two symmetric tongues of graft and sutured to the remnants of the aortic root wall. The dysplastic right coronary cusp was resected, and autologous pericardial patches augmented the deficiency of cusp tissue between the left and noncoronary cusps. A suture annuloplasty was used in 20 cases. All patients were followed clinically and echocardiographically at 3, 6, and 12 months and at yearly intervals. Cumulative follow-up was 677 months (mean, 27 ± 18 months). RESULTS: No early or late death occurred. Intraoperative echocardiography revealed minimal or no aortic regurgitation in all patients; at discharge, systolic mean gradient was 6 ± 3 mm Hg. There was no bleeding or thromboembolic event during the follow-up. One patient exhibited endocarditis and underwent reoperation. Two patients experienced relevant recurrent aortic regurgitation for limited suture dehiscence between the patch and the cusp and were reoperated on between 16 and 32 months postoperatively. One patient underwent biologic valve replacement, and two valves were re-repaired. At 5 years, freedom from reoperation and aortic valve replacement was 81% and 91%, respectively. CONCLUSIONS: In the presence of UAV and aortic root dilatation, the concept of valve bicuspidization and root remodeling can be applied with satisfactory hemodynamic results. The hemodynamic function of an aortic valve preserved by this concept is good. If sufficient stability can be achieved, aortic valve replacement can be avoided in young patients with aortic regurgitation caused by UAV and root aneurysm.

Bicuspidization of the unicuspid aortic valve: a new reconstructive approach.

BACKGROUND: Unicuspid anatomy of the aortic valve is infrequent but may require intervention by age 40 for severe regurgitation. We propose a new repair technique for the regurgitant unicuspid valve by converting it into a bicuspid aortic valve. METHODS: Between November 2003 and September 2007, 20 patients underwent regurgitant unicuspid aortic valve repair: 13 had aortic regurgitation (AR) and 7 had combined regurgitation and stenosis. Four patients had previously undergone balloon valvuloplasty for critical aortic stenosis. The aim of the repair was to construct a bicuspid valve with two normal commissures and unrestricted cusp motion. The fused cusp tissue was divided anteriorly and a new commissure of normal height was created. Noncoronary and right coronary cusps were extended with autologous pericardium. Concomitant operations included ascending aortic replacement in 7 and resection of subaortic stenosis in 1. RESULTS: No early or late deaths occurred. Intraoperative echocardiography revealed minimal or no AR in 19 patients. Follow-up was 4 to 47 months. One patient underwent valve re-repair for recurrent and progressive aortic regurgitation 3 years postoperatively. All other valves remained stable throughout the follow-up period. Freedom from relevant aortic insufficiency (> or = II) at 4 years was 77%; freedom from reoperation was 67%; and freedom from valve replacement was 100%. CONCLUSIONS: The regurgitant unicuspid aortic valve can be repaired successfully and reproducibly by converting it into bicuspid anatomy. The functional results are comparable with those obtained in reconstructed bicuspid aortic valves. With this approach, replacement can be avoided in most patients with regurgitant unicuspid aortic valves.