Hybrid minigene splicing assay verified the pathogenicity of a novel splice site variant in the dystrophin gene of a Chinese patient with typical Duchenne muscular dystrophy phenotype

Wang, Zhihong; Lin, Yuan; Qiu, Liping; Zheng, Deyou; Yan, Ai-zhen; Zeng, Jian; Lan, Fenghua

doi:10.1515/cclm-2015-1042

Cited by 7 publications

(6 citation statements)

References 20 publications

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“…Therefore, transcriptomic analysis of the patient's muscle biopsy or functional studies (Minigenes), are required to confirm this prediction. 29 We did not identify missense mutations in our sample of 69 previously clinically diagnosed patients. According to the literature, this type of mutation is the less frequent within DMD/BMD patients, which was also supported by our results.…”

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confidence: 73%

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Genetic Profile of the Dystrophin Gene Reveals New Mutations in Colombian Patients Affected with Muscular Dystrophinopathy

Triana-Fonseca¹,

Parada-Márquez²,

Silva-Aldana³

et al. 2021

TACG

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Background Duchenne and Becker muscular dystrophies (DMD/BMD) are the most common human dystrophinopathies with recessive X-linked inheritance. Dystrophin gene deletions and duplications are the most common mutations, followed by point mutations. The aim of this study is to characterize the mutational profile of the dystrophin gene in Colombian patients with DMD/BMD. Material and Methods Mutational profiling was determined in 69 affected patients using Sanger sequencing, next-generation sequencing (NGS) and/or multiplex ligation dependent-probes amplification (MLPA). Genetic variants were classified according to molecular consequence and new variants were determined through database and literature analysis. Results Mutational profile in affected patients revealed that large deletions/duplications analyzed by MLPA accounted for 72.5% of all genetic variations. By using Sanger sequencing or NGS, we identified point mutations in 15.9% and small deletions in 11.6% of the patients. New mutations were found, most of them were point mutations or small deletions (10.1%). Conclusion Our results described the genetic profile of the dystrophin gene in Colombian patients with DMD and contribute to efforts to identify molecular variants in Latin American populations. For our population, 18.8% of cases could be treated with FDA or MDA approved molecular therapies based on specific mutations. These data contribute to the establishment of appropriate genetic counseling and potential treatment.

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confidence: 73%

“…Therefore, transcriptomic analysis of the patient’s muscle biopsy or functional studies (Minigenes), are required to confirm this prediction. 29 …”

Section: Discussionmentioning

confidence: 99%

Genetic Profile of the Dystrophin Gene Reveals New Mutations in Colombian Patients Affected with Muscular Dystrophinopathy

Triana-Fonseca¹,

Parada-Márquez²,

Silva-Aldana³

et al. 2021

TACG

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“…The inclusion of intronic sequences can disrupt the reading frame and generate stop codons. 22 Intronic mutations detected by NGS have been reported in a number of cases of DMD, [51][52][53][54] and muscle biopsies also have been used to confirm the presence of aberrant dystrophin mRNA in patients with DMD. 36,39,[55][56][57] In addition to cases of deep intronic mutations, muscle biopsies may also be considered if the patient presents with a discordant phenotype (ie, the genotype would predict DMD, but the patient presents with BMD, or vice versa) 22,58,59 ; however, obtaining this information will not ultimately change the disease course or how the patient is managed.…”

Section: Discussionmentioning

confidence: 99%

“…A + = 73%; A = 27% Strongly in favor Low 1C 3,11,12,35,43,[45][46][47][48][49][50][51][52][53][54] and expert opinion Statement 9: Delays in the initial clinical diagnosis/referral to a specialist, the sequential nature of the genetic testing process, and incomplete or nonexhaustive genetic testing should be addressed in order to prevent delays in reaching a complete genetic diagnosis for patients with DMD.…”

Section: The Journal Of Pediatrics • Wwwjpedscommentioning

confidence: 99%

Evidence-Based Consensus and Systematic Review on Reducing the Time to Diagnosis of Duchenne Muscular Dystrophy

Aartsma‐Rus

Hegde

Ben‐Omran

et al. 2019

The Journal of Pediatrics

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Section 1: Reducing the time to diagnosis of DMD Statement 1: The following signs, symptoms, and characteristics should be considered typical indicators of DMD: calf hypertrophy (pseudohypertrophy); delayed walking; difficulty climbing/descending stairs; difficulty rising from the floor; difficulty running/walking; elevated serum CK levels (including elevated ALT and AST); a family history of DMD; frequent falls; Gowers' sign; male sex; and muscle weakness. ALT, alanine aminotransferase; AST, aspartate aminotransferase; CGH, comparative genome hybridization; CK, creatine kinase; DMD, Duchenne muscular dystrophy; GRADE, Grading of Recommendations Assessment, Development, and Evaluation. *The level of evidence for most of the statements was graded as either low or moderate, owing to the fact that most of the studies included here are observational in nature rather than randomized controlled trials (due to the nature of this initiative). When there were multiple corroborative supporting observational studies, we have selected "moderate" for quality of evidence. †Consensus: A + = strongly agree; A = agree; N = neither agree nor disagree; D = disagree; D + = strongly disagree. Grade of recommendation: 1A = strong recommendation, high-quality evidence; 1B = strong recommendation, moderate-quality evidence; 1C = strong recommendation, low-quality or very low-quality evidence; 2A = weak recommendation, high-quality evidence; 2B = weak recommendation, moderate-quality evidence; 2C = weak recommendation, low-quality or very low-quality evidence. ‡Where applicable depending on country-specific legislation on presymptomatic testing of patients aged ≤18 years.

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“…It was a pity that the muscle for RNA samples of the proband was degraded and was unfeasible to design a hybrid minigene to transfection of HeLa cells for splicing assay because of the huge intron 1. [ 24 ] So, we could not confirm the splicing mutation, but only assume these 2 different intron retentions and a new splice site from the RNA transcript, through computational prediction. It has deleterious effect on the precursor (pre) mRNAs splicing.…”

Section: Discussionmentioning

confidence: 99%

A novel DMD splicing mutation found in a family responsible for X-linked dilated cardiomyopathy with hyper-CKemia

Tang

Song²,

Ji³

et al. 2018

Medicine

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This study was aimed to detect a new mutation responsible for X-linked dilated cardiomyopathy with hyper-CKemia.We studied a proband who presented with cardiac symptoms with hyper-CKemia, but no clinical skeletal involvement in physical examination, laboratory tests, electromyography, echocardiography, and magnetic resonance imaging (MRI) of cardiac muscles. Muscle biopsy for histopathology and immunohistochemistry for accessing sarcolemma changes. The next-generation sequencing and bioinformatics analysis were performed on the patient and Sanger sequencing was confirmed on the other 6 unaffected families.The clinic investigations illustrated a dilated cardiomyopathy. Histopathology and immunohistochemistry showed dystrophic changes and an obvious reduction of dystrophin-N and δ-sarcoglycan, respectively. One hemizygous splicing pathogenic mutation c.31 + 1G > C of exon 1 in the DMD gene (chrX33229398, NM_00 4006) was finally identified in the patient and his nephew, but it was carried in his mother and sister.A novel small mutation was identified at the first exon-intron boundary splicing site by next-generation sequencing and bioinformatics analysis.

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Hybrid minigene splicing assay verified the pathogenicity of a novel splice site variant in the dystrophin gene of a Chinese patient with typical Duchenne muscular dystrophy phenotype

Abstract: The novel splice site mutation caused DMD in the proband by aberrant splicing. We suggested that combined applications of MLPA, NGS, HMSA and bioinformatics are comprehensive and effective methods for diagnosis and aberrant splicing study of DMD.

Cited by 7 publications

References 20 publications

Genetic Profile of the Dystrophin Gene Reveals New Mutations in Colombian Patients Affected with Muscular Dystrophinopathy

Genetic Profile of the Dystrophin Gene Reveals New Mutations in Colombian Patients Affected with Muscular Dystrophinopathy

Evidence-Based Consensus and Systematic Review on Reducing the Time to Diagnosis of Duchenne Muscular Dystrophy

A novel DMD splicing mutation found in a family responsible for X-linked dilated cardiomyopathy with hyper-CKemia

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