Compound heterozygous c.598_612del and c.1746-20C\u2009>\u2009G CAPN3 genotype cause autosomal recessive limb-girdle muscular dystrophy-1: a case report

Evelina Siavrienė,Violeta Mikštienė,Gunda Petraitytė,Aušra Morkūnienė,Tautvydas Rančelis,Eglė Preikšaitienė,Vaidutis Kučinskas,Algirdas Utkus,Birutė Burnytė

doi:10.1186/s12891-021-04920-3

Evelina Siavrienė, Violeta Mikštienė + Show 7 more

Open Access

https://doi.org/10.1186/s12891-021-04920-3

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Abstract

BackgroundAutosomal recessive limb–girdle muscular dystrophy-1 (LGMDR1), also known as calpainopathy, is a genetically heterogeneous disorder characterised by progression of muscle weakness. Homozygous or compound heterozygous variants in the CAPN3 gene are known genetic causes of this condition. The aim of this study was to confirm the molecular consequences of the CAPN3 variant NG_008660.1(NM_000070.3):c.1746-20C > G of an individual with suspected LGMDR1 by extensive complementary DNA (cDNA) analysis.Case presentationIn the present study, we report on a male with proximal muscular weakness in his lower limbs. Compound heterozygous NM_000070.3:c.598_612del and NG_008660.1(NM_000070.3):c.1746-20C > G genotype was detected on the CAPN3 gene by targeted next-generation sequencing (NGS). To confirm the pathogenicity of the variant c.1746-20C > G, we conducted genetic analysis based on Sanger sequencing of the proband’s cDNA sample. The results revealed that this splicing variant disrupts the original 3′ splice site on intron 13, thus leading to the skipping of the DNA fragment involving exon 14 and possibly exon 15. However, the lack of exon 15 in the CAPN3 isoforms present in a blood sample was explained by cell-specific alternative splicing rather than an aberrant splicing mechanism. In silico the c.1746-20C > G splicing variant consequently resulted in frameshift and formation of a premature termination codon (NP_000061.1:p.(Glu582Aspfs*62)).ConclusionsBased on the results of our study and the literature we reviewed, both c.598_612del and c.1746-20C > G variants are pathogenic and together cause LGMDR1. Therefore, extensive mRNA and/or cDNA analysis of splicing variants is critical to understand the pathogenesis of the disease.

Highlights

Autosomal recessive limb–girdle muscular dystrophy-1 (LGMDR1), known as calpainopathy, is a genetically heterogeneous disorder characterised by progression of muscle weakness
Extensive mRNA and/or complementary DNA (cDNA) analysis of splicing variants is critical to understand the pathogenesis of the disease
The underlying pathological mechanisms involved in LGMDR1 are not well known, studies of Capn3 knockdown mice identified a human calpainopathy-like phenotype, indicating that this disease is caused by defects in the CAPN3 gene (MIM #114240), which is located on chromosome 15q15 [3]

Summary

Introduction

Autosomal recessive limb–girdle muscular dystrophy-1 (LGMDR1), known as calpainopathy, is a genetically heterogeneous disorder characterised by progression of muscle weakness. The heterozygous CAPN3 variants have been reported to cause autosomal dominant limb–girdle muscular dystrophy-4 (LGMDD4; MIM #618129; ORPHA # 102014), which has a later onset and milder phenotype than LGMDR1 [2, 4, 5]. Besides DNA sequence variants in the CAPN3 gene, it has been suggested that other genetic and environmental factors modulate the clinical phenotype of LGMDR1, because variable degrees of muscle involvement and disease progression have been reported in individuals with the same genetic variant [6]

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