Illegitimate transcription: its use for studying genetic abnormalities in lymphoblastoid cells from patients with Glanzmann thrombasthenia

Négrier, Claude; Vinciguerra, Christine; Attali, Olivier; C, Grenier; Larcher, Marie Estelle; Dechavanne, M

doi:10.1046/j.1365-2141.1998.00515.x

Cited by 15 publications

(11 citation statements)

References 29 publications

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“…The mRNA that was used for RT‐PCR was extracted from B‐cell lines. Although the same analysis could not be performed on the granulocytes of AGR patients, it is assumed that the amount of the specific mRNA in the B‐cell lines of patients and controls represents the proportional amount of mRNA present in any other cells where NQO2 is transcribed (30).…”

Section: Resultsmentioning

confidence: 99%

NQO2 gene is associated with clozapine‐induced agranulocytosis

et al. 2003

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Clozapine is a dibenzodiazepine neuroleptic with atypical pharmacological and clinical profiles. Treatment with this drug may be complicated with agranulocytosis (AGR). It is likely that defective oxidative mechanism may be the cause of AGR. A candidate gene, dihydronicotinamide riboside (NRH) quinone oxidoreductase 2 (NQO2), which is involved in detoxification of drugs, was selected. This gene has been mapped to the short arm of chromosome six. The gene was studied by single-strand conformation polymorphism analysis and direct sequencing in 98 schizophrenic patients that were treated with clozapine. Eighteen of these patients developed AGR. Ten polymorphisms in the coding regions, in intron 1, and in the promoter region were found, two of which were novel. Comparisons of the polymorphisms in the first intron in AGR patients and controls suggested that this site might be connected with AGR. Quantitative reverse transcriptase-polymerase chain reaction analysis showed that the level of NQO2 mRNA is low in AGR patients compared with the control group. Such a reduction in message suggests that the NQO2 gene may be involved in the development of clozapine-induced AGR.

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Section: Resultsmentioning

confidence: 99%

NQO2 gene is associated with clozapine‐induced agranulocytosis

et al. 2003

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“…12 and in exons 1 and 3 of the GPIIIa gene responsible for GT 17,18 were easily detected by DGGE analysis (data not shown). Moreover, mutations introduced in the promoter region of the GPIIb (a gift from Dr G. Uzan, Inserm U217, Grenoble, France) presented an altered melting behaviour with heteroduplex analysis (data not shown).…”

Section: Choice Of Experimental Conditionsmentioning

confidence: 87%

Description of 10 new mutations in platelet glycoprotein IIb (αIIb) and glycoprotein IIIa (β3) genes

et al. 2001

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In this study we have used denaturing gradient gel electrophoresis (DGGE) for identifying sequence alterations in glycoprotein (GP) IIb and IIIa genes from 20 patients affected by Glanzmann's thrombasthenia. These patients were from 16 different families. Using computer modelling, we divided the promoters, coding sequences and flanking splicing regions, in 31 segments for the GPIIb gene and 19 domains for the GPIIIa gene. We were able to find a mutation potentially affecting GPIIb-IIIa expression or function in 16 patients out of 20. In six patients from three families, the gypsy mutation modifying the splice donor site of intron 15 of the GPIIb gene was detected. In the other patients, 10 novel mutations were characterised, which were located either in the GPIIb gene (nine cases) or in the GPIIIa gene (one case). The type of mutation was nonsense mutation (one case), missense mutation (five cases), small insertion of 1 bp (one case) and splicing modifications (three cases). Among these genetic events, three were directly responsible for Glanzmann's thrombasthenia, four were localised in regions known to be involved in GPIIb-IIIa complex expression and three mutations were potentially responsible for Glanzmann's thrombasthenia.

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“…The use of LCLs has been previously proven to be instrumental in uncovering important pathogenic mechanisms in other diseases, such as hypertension, glycogen and lipid storage diseases, chronic granulomatous disease, hematologic disorders, and solid tumors (27)(28)(29)(30)(31)(32)(33). One illustrative example is essential hypertension.…”

Section: Discussionmentioning

confidence: 99%

Sphingosine kinase 1–mediated inhibition of Fas death signaling in rheumatoid arthritis B lymphoblastoid cells

Pi¹,

Tan²,

Hayes³

et al. 2006

Arthritis & Rheumatism

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Objective. It is becoming increasingly apparent that B cells play an important role in the pathogenesis of rheumatoid arthritis (RA). Due to the scarcity of B cells in RA, it has been technically difficult to functionally characterize B cell apoptosis in this disease. As a necessary first step to identify candidate aberrations, we investigated Fas-mediated signaling events in immortalized peripheral blood B lymphoblastoid cell lines (LCLs) from patients with RA and controls.Methods. Cell death was determined by the MTS assay, and apoptosis was detected by the TUNEL assay and DNA laddering. Proteolytic activation of caspase 3 was determined by immunoblotting, and its enzymatic activity was determined by a fluorometric technique. Messenger RNA (mRNA) expression was quantified by real-time polymerase chain reaction (PCR) analysis. The functional role of sphingosine kinase (SPHK) was determined by measuring its enzymatic activity, by quantifying the levels of its product, sphingosine 1-phosphate (S1P), and by investigating the ability of the SPHK inhibitor N,N-dimethylsphingosine and isozyme-specific small interfering RNA (siRNA) oligonucleotides to reverse signaling aberrations.Results. LCLs from patients with RA displayed disease-specific Fas-mediated signal transduction impairment with consequent resistance to cell death. RA LCLs displayed high constitutive SPHK activity and increased levels of S1P. Real-time PCR analysis showed higher SPHK-1 mRNA expression levels in RA patients compared with paired controls. Increased SPHK-1 (but not SPHK-2) mRNA levels were observed in synovial tissue from RA patients. Competitive inhibitors of SPHK reversed the resistance of RA LCLs to Fasinduced apoptosis. Additionally, resistance to Fasmediated signaling was reversed by siRNA oligonucleotides specific for SPHK-1 but not by oligonucleotides specific for SPHK-2.Conclusion. These findings demonstrate diseasespecific resistance to Fas-mediated death signaling in patients with RA and implicate increased SPHK-1 activity as the cause of this aberration.

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Illegitimate transcription: its use for studying genetic abnormalities in lymphoblastoid cells from patients with Glanzmann thrombasthenia

Cited by 15 publications

References 29 publications

NQO2 gene is associated with clozapine‐induced agranulocytosis

NQO2 gene is associated with clozapine‐induced agranulocytosis

Description of 10 new mutations in platelet glycoprotein IIb (αIIb) and glycoprotein IIIa (β3) genes

Sphingosine kinase 1–mediated inhibition of Fas death signaling in rheumatoid arthritis B lymphoblastoid cells

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