GM17726
LCL from B-Lymphocyte
Description:
MUSCULAR DYSTROPHY, LIMB-GIRDLE, TYPE 2A; LGMD2A
|
Repository
|
NIGMS Human Genetic Cell Repository
|
| Subcollection |
Heritable Diseases
Muscular Dystrophies |
| Class |
Congenital Muscle Diseases |
| Class |
Disorders with Trinucleotide Expansions |
|
Biopsy Source
|
Peripheral vein
|
|
Cell Type
|
B-Lymphocyte
|
|
Tissue Type
|
Blood
|
|
Transformant
|
Epstein-Barr Virus
|
|
Sample Source
|
LCL from B-Lymphocyte
|
|
Family Member
|
1
|
|
Relation to Proband
|
proband
|
|
Confirmation
|
Clinical summary/Case history
|
|
Species
|
Homo sapiens
|
|
Common Name
|
Human
|
|
Remarks
|
|
| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin Confirmed by Nucleoside Phosphorylase,Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis |
| |
| creatine kinase |
According to the submitter, biochemical test results for this subject showed increased enzyme activity. EC Number: 2.7.3.2 |
| |
| Gene |
CAPN3 |
| Chromosomal Location |
15q15.1-q21.1 |
| Allelic Variant 1 |
114240.0010; LIMB-GIRDLE MUSCULAR DYSTROPHY, TYPE 2A |
| Identified Mutation |
ARG490GLN |
| |
| Gene |
CAPN3 |
| Chromosomal Location |
15q15.1-q21.1 |
| Allelic Variant 2 |
THR184ARGfs*36; LIMB-GIRDLE MUSCULAR DYSTROPHY, TYPE 2A |
| Identified Mutation |
THR184ARGfs*36 |
| Remarks |
Clinically affected; progressive bilateral weakness of arms and legs; wheelchair-dependent; weak facial muscles; scapular winging; elevated serum CPK; no chromosome 4 contraction; normal methylation levels; D4Z4 repeats for this donor subject are as follows: at 4q35: 28/59 copies (clinically unaffected individuals usually have >10 D4Z4 repeats on both alleles at chromosome 4q35) and at 10q26: 5/22 copies; this patient presented with FSHD symptoms, but does not have typical D4Z4 repeats (an occurrence in about 5% of cases); for additional genetic and epigenetic characterization, D4Z4 allele sizes, subtelomeric polymorphisms and methylation were tested using pulsed-field gel electrophoresis and Southern blotting with non-radioactive digoxigenin-labeled probes (Leidenroth et al. Eur J Hum Genet, 2012,PMID 22378277); GM17726 has three A-type telomeres and four D4Z4 alleles - the two larger alleles are from chromosome 4 while two smaller alleles are from chromosome 10; the original diagnosis was indicated as non-4q FSHD; Leidenroth et al. (2012) used whole exome sequencing to identify two known pathogenic mutations in CAPN3: a heterozygous G>A transition resulting in a substitution (Arg490Gln) and a 1 bp deletion of adenine in exon 4 causing a frameshift (Thr184Argfs*36); RT-PCR and Sanger sequencing confirmed that the two mutations are on different chromosomes; this compound heterozygous mutation in CAPN3 indicates a diagnosis of LGMD2A and not FSHD. |
| Leidenroth A, Sorte HS, Gilfillan G, Ehrlich M, Lyle R, Hewitt JE, Diagnosis by sequencing: correction of misdiagnosis from FSHD2 to LGMD2A by whole-exome analysis European journal of human genetics : EJHG20:999-1003 2012 |
| PubMed ID: 22378277 |
| Split Ratio |
1:5 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Medium |
Roswell Park Memorial Institute Medium 1640 with 2mM L-glutamine or equivalent |
| Serum |
15% fetal bovine serum Not Inactivated |
| Substrate |
None specified |
| Subcultivation Method |
dilution - add fresh medium |
| Supplement |
- |
|