Use of RNAsequencing to detect abnormal transcription of the collagen α2 (VI) chain gene that can lead to Bethlem myopathy.
Int J Mol Med
; 47(3)2021 03.
Article
em En
| MEDLINE
| ID: mdl-33537799
Bethlem myopathy (BM) is an autosomal dominant or autosomal recessive disorder and is usually associated with mutations in the collagen VI genes. In the present study, the pathogenicity of a novel splicesite mutation was explored using RNAsequencing in a family with suspected BM, and a myopathy panel was performed in the proband. The genetic status of all family members was confirmed using Sanger sequencing. Clinical data and magnetic resonance imaging (MRI) features were also documented. In silico analysis was performed to predict the effects of the splice mutation. RNAsequencing and reverse transcription (RT)PCR were used to assess aberrant splicing. Immunocytochemistry was conducted to measure collagen VI protein levels within the gastrocnemius and in cultured skin fibroblasts. The results revealed that three patients in the family shared a similar classic BM presentation. MRI revealed distinct patterns of fatty infiltration in the lower extremities. A novel splicing mutation c.7361G>C in the collagen α2 (VI) chain (COL6A2) gene was found in all three patients. In silico analysis predicted that the mutation would destroy the normal splice acceptor site. RNAsequencing detected two abnormal splicing variants adjacent to the mutation site, and RTPCR confirmed the RNAsequencing findings. Furthermore, a defect in the collagen protein within cultured fibroblasts was detected using immunocytochemistry. The mutation c.7361G>C in the COL6A2 gene caused aberrant splicing and led to premature termination of protein translation. In conclusion, these findings may improve our knowledge of mutations of the COL6A2 gene associated with BM and demonstrated that RNAsequencing can be a powerful tool for finding the underlying mechanism of a diseasecausing mutations at a splice site.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Transcrição Gênica
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Mutação Puntual
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Contratura
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Sítios de Splice de RNA
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Colágeno Tipo VI
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RNA-Seq
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Distrofias Musculares
Tipo de estudo:
Prognostic_studies
Limite:
Adolescent
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Adult
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Humans
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Male
Idioma:
En
Revista:
Int J Mol Med
Ano de publicação:
2021
Tipo de documento:
Article