GM16028

GM16028 LCL from B-Lymphocyte

Description:

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

Affected:

No Data

Sex:

Female

Age:

64 YR (At Sampling)

Overview

Repository

NIGMS Human Genetic Cell Repository

Subcollection

Heritable Diseases

Class

Mutations of the Hemoglobin Loci

Biopsy Source

Peripheral vein

Cell Type

B-Lymphocyte

Tissue Type

Blood

Transformant

Epstein-Barr Virus

Sample Source

LCL from B-Lymphocyte

Race

White

Relation to Proband

proband

Confirmation

Molecular characterization before cell line submission to CCR

Species

Homo sapiens

Common Name

Human

Remarks

Deep vein thrombosis; Donor subject is heterozygous for the G-to-A transition at position 20210 of the prothrombin gene (20210G>A); 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)]; heterozygous for a G>A transition at nucleotide 1691 (1691G>A) of the Factor V (F5) gene resulting in a substitution of glutamine for arginine at codon 506 [Arg506Gln (R506Q)]; heterozygous for an A>T transversion at nucleotide 193 in exon 2 of the HFE (HLA-H) gene [193A>T] resulting in a substitution of cysteine for serine at codon 65 [Ser65Cys (S65C)]; family history: one daughter had postpartum thrombosis.

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 MTHFR mutation in this cell line has been verified by 5 laboratories. Methods used for mutation identification include: allele-specific amplification assay with gel electrophoresis; PCR + restriction endonuclease digestion and gel electrophoresis.

Gene

Chromosomal Location

1q23

Allelic Variant 1

227400.0001; THROMBOPHILIA DUE TO DEFICIENCY OF COFACTOR FOR ACTIVATED PROTEIN

Identified Mutation

20009404T>C; Bertina et al. [Nature 369: 64-67 (1994)] identified a mutation in the F5 gene as the basis of deficiency of the cofactor of activated protein C in a family with APC resistance and proneness to thrombosis. In 2 patients classified as homozygous for the deficiency of the cofactor, they found homozygosity for a guanine-to-adenine substitution at nucleotide 1691. This mutation predicted the replacement of arg506 (CGA) by gln (CAA). They referred to the mutation as FV Q506 or FV Leiden. (This mutation is also known as R506Q, using the single letter symbols for the amino acid change. It is also known as G1691A, or, to avoid confusion of the single letter symbol for nucleotides with similar symbols for amino acids, 1691G-A.)

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

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

HFE

Chromosomal Location

6p22.2

Allelic Variant 1

613609.0003; HEMOCHROMATOSIS

Identified Mutation

SER65CYS; Mura et al. [Blood 93: 2502-2505 (1999)] reported on the analysis of the cys282-to-tyr (C282Y; 235200.0001), his63-to-asp (H63D; 235200.0002), and ser65-to-cys (S65C) mutations of the HFE gene in a series of 711 probands with hereditary hemochromatosis and 410 controls. The results confirmed that the C282Y substitution is the main mutation involved in HH, accounting for 85% of carrier chromosomes, whereas the H63D substitution represented 39% of the HH chromosomes that did not carry the C282Y mutation. In addition, the screening showed that the S65C substitution, which results from a 193A-T transversion, was significantly enriched in probands with at least one chromosome without an assigned mutation. This substitution accounted for 7.8% of HH chromosomes that were neither C282Y nor H63D. This enrichment of S65C among HH chromosomes suggested that the S65C substitution is associated with a mild form of hemochromatosis.

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

Poort SR, Rosendaal FR, Reitsma PH, Bertina RM, A common genetic variation in the 3'-untranslated region of the prothrombin gene is associated with elevated plasma prothrombin levels and an increase in venous thrombosis. Blood88(10):3698-703 1996

PubMed ID: 8916933

Frosst P, Blom HJ, Milos R, Goyette P, Sheppard CA, Matthews RG, Boers GJ, den Heijer M, Kluijtmans LA, van den Heuvel LP, et al, A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase. Nat Genet10(1):111-3 1995

PubMed ID: 7647779

Bertina RM, Koeleman BP, Koster T, Rosendaal FR, Dirven RJ, de Ronde H, van der Velden PA, Reitsma PH, Mutation in blood coagulation factor V associated with resistance to activated protein C. Nature369(6475):64-7 1994

PubMed ID: 8164741