GM13591

GM13591 LCL from B-Lymphocyte

Description:

CYSTIC FIBROSIS; CF
CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR; CFTR
HEMOCHROMATOSIS; HFE
5,10-@METHYLENETETRAHYDROFOLATE REDUCTASE; MTHFR
HUMAN GENE MUTATION PANEL - HEMOCHROMATOSIS
HUMAN GENE MUTATION PANEL - CYSTIC FIBROSIS (VERSION 2)
HOMEOSTATIC IRON REGULATOR; HFE

Affected:

Yes

Sex:

Female

Age:

32 YR (At Sampling)

Overview

Repository

NIGMS Human Genetic Cell Repository

Subcollection

Heritable Diseases

Class

Other Disorders of Known Biochemistry

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 after cell line submission to CCR

Species

Homo sapiens

Common Name

Human

Remarks

Mild symptoms; donor subject is a compound heterozygote for mutations in the CFTR gene: allele one carries a deletion of codon 508 (CTT) in exon 10 which leads to deletion of phenylalanine-508 [PHE508DEL] and allele two carries a G-to-A transition at nucleotide 482 (482G>A) in exon 4, resulting in a change of arginine to histidine [ARG117HIS (R117H)]; analysis of a DNA variant in a noncoding region of the CFTR gene (polypyrimidine tract in intron 8) showed this donor has alleles 5T/9T; donor subject is homozygous 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

GENE MAPPING & DOSAGE STUDIES - Y CHROMOSOME

PCR analysis of DNA from this cell culture gave a negative result with a primer for Yq11, DYS227.

MUTATION VERIFICATION

The CFTR mutations in this cell line have been verified by 6 laboratories. Methods used for mutation identification include: commercial kits (ABI and Roche); electrophoresis for RFLP and size analysis; sequencing; mutation scanning (heteroduplex analysis, dHPLC, SSCP, DGGE); chip technology; oligonucleotide ligation assay.

CYSTIC FIBROSIS TRANSMEMBRANE CONDUCTANCE REGULATOR

The CFTR gene mutation data for this repository number was verified by sequencing.

Gene

CFTR

Chromosomal Location

7q31.2

Allelic Variant 1

602421.0001; CYSTIC FIBROSIS

Identified Mutation

PHE508DEL; Deletion of codon 508 (CTT) in exon 10 leads to deletion of phenylalanine-508 (delta-F508).

Gene

HFE

Chromosomal Location

6p22.2

Allelic Variant 1

613609.0002; HEMOCHROMATOSIS

Identified Mutation

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

HFE

Chromosomal Location

6p22.2

Allelic Variant 2

613609.0002; HEMOCHROMATOSIS

Identified Mutation

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

CFTR

Chromosomal Location

7q31.2

Allelic Variant 2

602421.0005; CYSTIC FIBROSIS

Identified Mutation

ARG117HIS; In 2 presumably unrelated families with mild CF, Dean et al. [Cell 61: 863-870 (1990)] found a G-to-A transition at nucleotide 482 in exon 4, resulting in a change of arginine to histidine (R117H).

Phenotypic Data

Remarks

Publications

Xia Y, Katz M, Chandramohan D, Bechor E, Podgursky B, Hoxie M, Zhang Q, Chertman W, Kang J, Blue E, Chen J, Schleede J, Slotnick NR, Du X, Boostanfar R, Urcia E, Behr B, Cohen J, Siddiqui N, The first clinical validation of whole-genome screening on standard trophectoderm biopsies of preimplantation embryos F&S reports5:63-71 2023

PubMed ID: 38524212

Leung ML, Watson DJ, Vaccaro CN, Mafra F, Wenocur A, Wang T, Hakonarson H, Santani A, Evaluating sequence data quality from the Swift Accel-Amplicon CFTR Panel Scientific data7:8 2019

PubMed ID: 31913291

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

Hadd AG, Laosinchai-Wolf W, Novak CR, Badgett MR, Isgur LA, Goldrick M, Walkerpeach CR, Microsphere bead arrays and sequence validation of 5/7/9T genotypes for multiplex screening of cystic fibrosis polymorphisms The Journal of molecular diagnostics : JMD6:348-55 2004

PubMed ID: 15507674

Pont-Kingdon G, Jama M, Miller C, Millson A, Lyon E, Long-range (177 kb) allele-specific polymerase chain reaction method for direct haplotyping of R117H and IVS-8 mutations of the cystic fibrosis transmembrane regulator gene The Journal of molecular diagnostics : JMD6:264-70 2004

PubMed ID: 15269305

Sebastian S, Spitzer SG, Grosso LE, Amos J, Schaefer FV, Lyon E, Wolff DJ, Hajianpour A, Taylor AK, Millson A, Stenzel TT, Multicenter characterization and validation of the intron-8 poly(T) tract (IVS8-T) status in 25 Coriell cell repository cystic fibrosis reference cell lines for cystic fibrosis transmembrane conductance regulator (CFTR) gene mutation assays. Clin Chem50(1):251-4 2004

PubMed ID: 14709668

Bernacki SH, Farkas DH, Shi W, Chan V, Liu Y, Beck JC, Bailey KS, Pratt VM, Monaghan KG, Matteson KJ, Schaefer FV, Friez M, Shrimpton AE, Stenzel TT, Bioelectronic sensor technology for detection of cystic fibrosis and hereditary hemochromatosis mutations. Arch Pathol Lab Med127(12):1565-72 2003

PubMed ID: 14632577

Dunbar SA, Jacobson JW, Application of the luminex LabMAP in rapid screening for mutations in the cystic fibrosis transmembrane conductance regulator gene: A pilot study. Clin Chem46(9):1498-500 2000

PubMed ID: 10973900