GM16000

GM16000 LCL from B-Lymphocyte

Description:

COAGULATION FACTOR II; F2
HEMOCHROMATOSIS; HFE
5,10-@METHYLENETETRAHYDROFOLATE REDUCTASE; MTHFR
HUMAN GENE MUTATION PANEL - DISORDERS OF THROMBOSIS
HUMAN GENE MUTATION PANEL - HEMOCHROMATOSIS
HOMEOSTATIC IRON REGULATOR; HFE

Affected:

Yes

Sex:

Female

Age:

33 YR (At Sampling)

Overview

Repository

NIGMS Human Genetic Cell Repository

Subcollection

Heritable Diseases

Class

Disorders of Uncertain Biochemical Etiology

Biopsy Source

Peripheral vein

Cell Type

B-Lymphocyte

Tissue Type

Blood

Transformant

Epstein-Barr Virus

Sample Source

LCL from B-Lymphocyte

Race

White

Country of Origin

USA

Relation to Proband

proband

Confirmation

Clinical summary/Case history

Species

Homo sapiens

Common Name

Human

Remarks

Clinically affected; hyperprothrombinemia; donor subject is homozygous for the G-to-A transition at position 20210 of the prothrombin gene (20210G>A); donor subject is also heterozygous for a C>G transversion at nucleotide 187 in exon 2 of the HFE (HLA-H) gene [187C>G] resulting in a substitution of aspartic acid for histidine at codon 63 [His63Asp (H63D)]; donor subject is heterozygous for a C>T mutation at nucleotide 677 in exon 4 of the methylenetetrahydrofolate reductase (MTHFR) gene [677C>T] that results in a substitution of a valine for an alanine at codon 222 [Ala222Val (A222V)]

Characterizations

IDENTIFICATION OF SPECIES OF ORIGIN

Species of Origin Confirmed by Nucleoside Phosphorylase,Glucose-6-Phosphate Dehydrogenase, and Lactate Dehydrogenase Isoenzyme Electrophoresis

MUTATION VERIFICATION

The prothrombin mutations in this cell line have been verified by 5 laboratories. Methods used for mutation identification include: PCR with mismatched primer introducing allele-specific restriction enzyme site and gel electrophoresis; PCR + restriction endonuclease digestion and gel electrophoresis; Invader assay.

Gene

Chromosomal Location

11p11-q12

Allelic Variant 1

176930.0009; HYPERPROTHROMBINEMIA WITH RISK OF THROMBOSIS

Identified Mutation

20210G>A; Poort et al. (1996) described a common genetic variation in the 3-prime untranslated region of the prothrombin gene that is associated with elevated plasma prothrombin levels and an increased risk of venous thrombosis: a G-to-A transition at position 20210 Degen and Davie, 1987. They found this single base substitution in 18% of probands of thrombophilic families, 6% of unselected consecutive patients with deep-vein thrombosis, and 2% of healthy controls. Rosendaal et al. (1997) found that the mutation was associated with a 4-fold increased risk of myocardial infarction in women, while among men the risk was increased 1.5-fold Doggen et al., 1998. Rosendaal et al. (1998) presented data from 11 centers and 9 countries, representing a total of 5,527 tested individuals. Among these, 111 heterozygous carriers of the 20210A mutation were found. The overall prevalence estimate was 2.0%. In southern Europe, the prevalence was 3.0%, nearly twice as high as the prevalence in northern Europe (1.7%). The prothrombin variant appeared to be very rare in individuals of Asian and African descent.

Gene

HFE

Chromosomal Location

6p22.2

Allelic Variant 1

613609.0002; HEMOCHROMATOSIS

Identified Mutation

c.187C>G (p.HIS63ASP); A mutation caused by a C-to-G transversion in exon 2 results in a histidine to aspartic acid substitution at codon position 63 [his63asp (H63D)] in the HFE gene.

Gene

MTHFR

Chromosomal Location

1p36.3

Allelic Variant 1

607093.0003; MTHFR THERMOLABILE POLYMORPHISM

Identified Mutation

677C>T; Frosst et al. [Nature Genet. 10: 111-113 (1995)] identified a C-to-T substitution at nucleotide 677 that converted an alanine to a valine residue. The alteration created a HinfI site that was used to screen 114 unselected French-Canadian chromosomes; the allele frequency of the substitution was 0.38. The mutation in the heterozygous or homozygous state correlated with reduced enzyme activity and increased thermolability in lymphocyte extracts; in vitro expression of the mutagenized cDNA containing the mutation confirmed its effect on thermolability of MTHFR. Individuals homozygous for the mutation had significantly elevated plasma homocysteine levels. Thus, the 677C-T mutation may represent an important genetic risk factor in vascular disease.

Gene

Chromosomal Location

11p11-q12

Allelic Variant 2

176930.0009; HYPERPROTHROMBINEMIA WITH RISK OF THROMBOSIS

Identified Mutation

Phenotypic Data

Remarks

Publications

Bao YP, Huber M, Wei TF, Marla SS, Storhoff JJ, Müller UR, SNP identification in unamplified human genomic DNA with gold nanoparticle probes Nucleic acids research33:e15 2005

PubMed ID: 15659576

Bernacki SH, Beck JC, Muralidharan K, Schaefer FV, Shrimpton AE, Richie KL, Levin BC, Pont-Kingdon G, Stenzel TT., Characterization of publicly available lymphoblastoid cell lines for disease-associated mutations in 11 genes. Clin Chem51(11):2156-9 2005

PubMed ID: 16244288

Moser MJ, Marshall DJ, Grenier JK, Kieffer CD, Killeen AA, Ptacin JL, Richmond CS, Roesch EB, Scherrer CW, Sherrill CB, Van Hout CV, Zanton SJ, Prudent JR, Exploiting the enzymatic recognition of an unnatural base pair to develop a universal genetic analysis system. Clin Chem49(3):407-14 2003

PubMed ID: 12600952