A Larger Spectrum of Intragenic Short Tandem Repeats Improves Linkage Analysis and Localization of Intragenic Recombination Detection in the Dystrophin Gene

Carsana, Antonella; Frisso, Giulia; Tremolaterra, Maria Roberta; Ricci, Elisabetta; Rasmo, Domenico De; Salvatore, Francesco

doi:10.2353/jmoldx.2007.060056

Cited by 18 publications

(8 citation statements)

References 21 publications

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“…Human samples to perform genetic analysis were obtained following informed consent of patients according to the Declaration of Helsinki. Genomic DNA was isolated from peripheral whole blood as previously described 29 . All coding exons, and 5′ and 3′ UTRs of genes involved in HCM were amplified by PCR and analysed by automatic sequencing using previously reported protocols 9 .…”

Section: Molecular Genetics Of Hcm Patientsmentioning

confidence: 99%

Protein Thermodynamic Destabilization in the Assessment of Pathogenicity of a Variant of Uncertain Significance in Cardiac Myosin Binding Protein C

Pricolo

Herrero-Galán

Mazzaccara

et al. 2020

J. of Cardiovasc. Trans. Res.

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In the era of Next Generation Sequencing (NGS), genetic testing for inherited disorders identifies an ever-increasing number of variants whose pathogenicity remains unclear. These variants of uncertain significance (VUS) limit the reach of genetic testing in clinical practice. The VUS for Hypertrophic Cardiomyopathy (HCM), the most common familial heart disease, constitute over 60% of entries for missense variants shown in ClinVar database. We have studied a novel VUS (c.1809T>G-p.I603M) in the most frequently mutated gene in HCM, MYBPC3, which codes for cardiac myosin-binding protein C (cMyBPC). Our determinations of pathogenicity integrate bioinformatics evaluation and functional studies of RNA splicing and protein thermodynamic stability. In silico prediction and mRNA analysis indicated no alteration of RNA splicing induced by the variant. At the protein level, the p.I603M mutation maps to the C4 domain of cMyBPC. Although the mutation does not perturb much the overall structure of the C4 domain, the stability of C4 I603M is severely compromised as detected by circular dichroism and differential scanning calorimetry experiments. Taking into account the highly destabilizing effect of the mutation in the structure of C4, we propose reclassification of variant p.I603M as likely pathogenic. Looking into the future, the workflow described here can be used to refine the assignment of pathogenicity of variants of uncertain significance in MYBPC3.

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Section: Molecular Genetics Of Hcm Patientsmentioning

confidence: 99%

Protein Thermodynamic Destabilization in the Assessment of Pathogenicity of a Variant of Uncertain Significance in Cardiac Myosin Binding Protein C

Pricolo

Herrero-Galán

Mazzaccara

et al. 2020

J. of Cardiovasc. Trans. Res.

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“…Effective treatments are limited for DMD patients, and research for genetic-based therapies is ongoing [7]. As a consequence, the analysis of the DMD gene is of utmost importance for the identification of the underlying molecular defect, because it can confirm the clinical diagnosis, reveal patients' genotype, address patients to the most opportune therapeutic options, and allow the identification of carrier females and the application of prenatal tests [8][9][10].…”

Section: Introductionmentioning

confidence: 99%

Metabolic Alterations in Cardiomyocytes of Patients with Duchenne and Becker Muscular Dystrophies

Esposito

Carsana

2019

JCM

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Duchenne and Becker muscular dystrophies (DMD/BMD) result in progressive weakness of skeletal and cardiac muscles due to the deficiency of functional dystrophin. Respiratory failure is a leading cause of mortality in DMD patients; however, improved management of the respiratory symptoms have increased patients’ life expectancy, thereby also increasing the clinical relevance of heart disease. In fact, the prevalence of cardiomyopathy, which significantly contributes to mortality in DMD patients, increases with age and disease progression, so that over 95% of adult patients has cardiomyopathy signs. We here review the current literature featuring the metabolic alterations observed in the dystrophic heart of the mdx mouse, i.e., the best-studied animal model of the disease, and discuss their pathophysiological role in the DMD heart. It is well assessed that dystrophin deficiency is associated with pathological alterations of lipid metabolism, intracellular calcium levels, neuronal nitric oxide (NO) synthase localization, and NO and reactive oxygen species production. These metabolic stressors contribute to impair the function of the cardiac mitochondrial bulk, which has a relevant pathophysiological role in the development of cardiomyopathy. In fact, mitochondrial dysfunction becomes more severe as the dystrophic process progresses, thereby indicating it may be both the cause and the consequence of the dystrophic process in the DMD heart.

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Section: Case Study: Clinical Molecular Biology Investigationmentioning

confidence: 99%

“…Informed written consent for DNA analysis was obtained from the proband, his parents, and his younger brother, according to the procedure established by the Second Helsinki Declaration and according to Italian and local Regulations (Ethic Committee of the Monaldi Hospital-Naples-#189 15/03/2009). Genomic DNA was isolated from peripheral whole blood, as described elsewhere [7]. The patient's DNA was amplified by polymerase chain reaction (PCR) for all exons, the flanking regions ranged from a minimum of 25 bp, and a maximum of 220 bp of the six following most often involved genes in arrhythmogenic right ventricular cardiomyopathy (ARVC) disease: Plakophilin (PKP2), Desmoplakin (DSP), Desmoglein-2 (DSG2), Desmocollin-2 (DSC2), Ryanodine Receptor 2 (RYR2), Junction Plakoglobin (JUP), using previously reported protocols [8].…”

Section: Case Study: Clinical Molecular Biology Investigationmentioning

confidence: 99%

Genotype-Phenotype Correlation: A Triple DNA Mutational Event in a Boy Entering Sport Conveys an Additional Pathogenicity Risk

Limongelli

Nunziato

Mazzaccara

et al. 2020

Genes

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The purpose of this paper is to present a clinical and laboratory study of a family, in which a 12-year-old boy was examined to assess his health status before starting competitive sports. A variety of clinical and instrumental tests were used to evaluate the status of the heart and its functions. Using Sanger sequencing (SS), we sequenced six related genes to verify suspected arrhythmogenic right ventricular cardiomyopathy (ARVC) hypothesized at the cardiac assessment and, subsequently, by a next-generation sequencing (NGS)-based multi-gene panel for more paramount genetic risk of sudden cardiac death (SCD) assessment. SS revealed two variants in the PKP2 gene, one was inherited from the father and the other from the mother. The analysis on a large panel of genes (n = 138), putatively associated with sudden cardiac death, revealed, in the proband, a third variant in a different gene (DES) that encodes the protein desmin. Our results indicate that: i) NGS revealed a mutational event in a gene not conventionally screened as a first-line test in the presence of clinical suspicion of the arrhythmic disease; ii) a plurality of variants in different genes in the same subject (the proband) may increase the risk of heart disease; iii) in silico analysis with various methodological software and bioinformatic prediction tools indicates that the cumulative effects of the three variants in the same subject constitute an additional risk factor. This case report indicates that more pathogenic variants or likely pathogenic variants can contribute to the clinical phenotype of an individual, thereby contributing to the diagnosis and prognosis of inherited heart diseases.

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A Larger Spectrum of Intragenic Short Tandem Repeats Improves Linkage Analysis and Localization of Intragenic Recombination Detection in the Dystrophin Gene

Cited by 18 publications

References 21 publications

Protein Thermodynamic Destabilization in the Assessment of Pathogenicity of a Variant of Uncertain Significance in Cardiac Myosin Binding Protein C

Protein Thermodynamic Destabilization in the Assessment of Pathogenicity of a Variant of Uncertain Significance in Cardiac Myosin Binding Protein C

Metabolic Alterations in Cardiomyocytes of Patients with Duchenne and Becker Muscular Dystrophies

Genotype-Phenotype Correlation: A Triple DNA Mutational Event in a Boy Entering Sport Conveys an Additional Pathogenicity Risk

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