GM27200
LCL from B-Lymphocyte
Description:
CEREBRAL ARTERIOPATHY, AUTOSOMAL DOMINANT, WITH SUBCORTICAL INFARCTS AND LEUKOENCEPHALOPATHY, TYPE 1; CADASIL1
NOTCH, DROSOPHILA, HOMOLOG OF, 3; NOTCH3
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Repository
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NIGMS Human Genetic Cell Repository
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| Subcollection |
Heritable Diseases
PIGI Consented Sample |
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Biopsy Source
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Peripheral vein
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Cell Type
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B-Lymphocyte
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Tissue Type
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Blood
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Transformant
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Epstein-Barr Virus
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Sample Source
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LCL from B-Lymphocyte
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Race
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White
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Ethnicity
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Not Hispanic/Latino
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Country of Origin
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USA
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Family History
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N
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Relation to Proband
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proband
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Confirmation
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Molecular characterization before cell line submission to CCR
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Species
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Homo sapiens
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Common Name
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Human
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Remarks
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| IDENTIFICATION OF SPECIES OF ORIGIN |
Species of Origin Confirmed by LINE assay |
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| Gene |
NOTCH3 |
| Chromosomal Location |
19p13.12 |
| Allelic Variant 1 |
600276.8; CADASIL |
| Identified Mutation |
ARG133CYS; In 3 unrelated patients with CADASIL1 (125310), Joutel et al. (1997) identified a 475C-T transition in exon N4 of the NOTCH3 gene, resulting in an arg133-to-cys (R133C) substitution in the EGF3 domain.
Mykkanen et al. (2004) performed haplotype analysis in 60 patients from 18 Finnish CADASIL families with the R133C mutation. Using 10 microsatellite markers, the authors found a similar haplotype linked to the mutation in all 18 pedigrees, indicating a single common ancestor for all of the Finnish R133C families. Age analysis of the founder mutation placed the introduction of the mutation in the late 1600s or early 1700s.
Opherk et al. (2009) showed that both wildtype and CADASIL-mutated (R133C) NOTCH3 receptor spontaneously formed oligomers and higher order multimers in vitro and that multimerization was mediated by disulfide bonds. CADASIL-associated mutations significantly enhanced multimerization compared with wildtype. Opherk et al. (2009) argued for a neomorphic effect of CADASIL mutations |
| Remarks |
Clinically affected; diagnosed by a neurologist at 46 years of age; frequent migraine headaches with aura; cerebrovascular disease; encephalopathy; cerebral infarction; increasing forgetfulness; depression; brain MRI showed no evidence of recent ischemic injury or abnormal restriction and no intracranial abnormality, but findings support the diagnosis of CADASIL: anterior subcortical temporal lobe signal alteration, signal changes within the external capsules and involvement of the bilateral thalami, portions of the basal ganglia and early changes within the pons; next-generation sequencing of whole blood genomic DNA revealed two heterozygous missense mutations in the Notch3 gene: autosomal dominant pathogenic c.397C>T (p.Arg133Cys) and autosomal VOUS c.386C>T (p.Ala129Val); medications and treatments: Prodrin (for migraines), Aleve, co-enzyme Q-10, Lipitor, Butterbur, fish oil, glucosamine. |
| Split Ratio |
1:5 |
| Temperature |
37 C |
| Percent CO2 |
5% |
| Percent O2 |
AMBIENT |
| 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 |
- |
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