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Genetic basis of cystinosis in Tunisian patients: Identification of novel mutation in CTNS gene.

Chkioua L, Khedhiri S, Grissa O, Aloui C, Turkia HB, Ferchichi S, Miled A, Froissart R, Acquaviva C, Laradi S - Meta Gene (2015)

Bottom Line: Nephropathic cystinosis (NC) is an autosomal recessive disorder characterized by defective transport of cystine across the lysosomal membrane and resulting in renal, ophthalmic, and other organ abnormalities.In addition, eight polymorphisms were identified in the 3 patients and their parents.This study expands the mutational and population spectrum of NC, representing the first molecular diagnosis of NC in Tunisian population.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biochemistry, F. Hached Hospital, 4000 Sousse, Tunisia ; University of Monastir, 5000 Monastir, Tunisia.

ABSTRACT
Nephropathic cystinosis (NC) is an autosomal recessive disorder characterized by defective transport of cystine across the lysosomal membrane and resulting in renal, ophthalmic, and other organ abnormalities. Mutations in the CTNS gene cause a deficiency of the transport protein, cystinosin. This study was performed to investigate mutations of the CTNS gene in three Tunisian families with NC. Polymerase chain reaction (PCR), ARMS multiplex PCR and direct sequencing were performed for molecular characterization of the CTNS gene in 3 unrelated Tunisian patients and their parents. Based on family history, prenatal diagnosis (PND) was performed in fetal DNA isolated from chorionic villi obtained at 10-12  weeks of gestation. None of the patients showed the most common 57-kb deletion in heterozygous or homozygous status. One patient was homozygous for the previously reported mutation c.1515G > A (p.G308R). One patient presented the novel gross deletion of 20,327 bp. One was homozygote for the previously reported mutation c.771_793del (p.Gly258Serfs*30). In addition, eight polymorphisms were identified in the 3 patients and their parents. The prenatal diagnosis in one family showed that the fetus DNA was heterozygous for the c.771_793del (p.Gly258Serfs*30) mutation. This study expands the mutational and population spectrum of NC, representing the first molecular diagnosis of NC in Tunisian population. The mutation screening of the CTNS gene was used for prenatal diagnosis to prevent and/or limit this inheritable disease in our country where the families are particularly large and have a high rate of consanguinity.

No MeSH data available.


Related in: MedlinePlus

Direct sequencing of CTNS gene in Tunisian patients.A: Sequence electropherograms of exon 11 of the CTNS gene for c.1515G > A (p.G308R) mutation.B: Sequence electropherograms of exon 10 of the CTNS gene c.771_793del (p.Gly258Serfs*30) mutation.C: Characterization of the large rearrangement removing exons 1 to 5 of the CTNS gene and a part of the intron 1 of CARKL gene.1C: Schematic representation of the CTNS and CARKL genes rearrangement. Blue boxes represent SGSH exons and purple boxes represent CARLK exons. The 20,327 bp deleted region is delineated. The black arrows correspond to the primers (IVS1F_CARKL/IVS6R_CTNS) used to amplify the junction fragment.PCR of the deletion breakpoint. Lane 1: DNA 100-bp increment ladder; Lane 2: patient; Lane 3: patient; Lane 4: patient's mother; and Lane 5: control.The amplification showed a 540 bp product corresponding to the junction fragment in the patients and her mother. This product was not present in the control DNA sample.
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f0015: Direct sequencing of CTNS gene in Tunisian patients.A: Sequence electropherograms of exon 11 of the CTNS gene for c.1515G > A (p.G308R) mutation.B: Sequence electropherograms of exon 10 of the CTNS gene c.771_793del (p.Gly258Serfs*30) mutation.C: Characterization of the large rearrangement removing exons 1 to 5 of the CTNS gene and a part of the intron 1 of CARKL gene.1C: Schematic representation of the CTNS and CARKL genes rearrangement. Blue boxes represent SGSH exons and purple boxes represent CARLK exons. The 20,327 bp deleted region is delineated. The black arrows correspond to the primers (IVS1F_CARKL/IVS6R_CTNS) used to amplify the junction fragment.PCR of the deletion breakpoint. Lane 1: DNA 100-bp increment ladder; Lane 2: patient; Lane 3: patient; Lane 4: patient's mother; and Lane 5: control.The amplification showed a 540 bp product corresponding to the junction fragment in the patients and her mother. This product was not present in the control DNA sample.

Mentions: Analysis of the entire CTNS gene in all patients and their parents revealed the presence of three mutations (Table 3): Two of these mutations were previously described c.1515G > A (p.G308R) in exon 11 of CTNS gene in patient TN6 of family 3 (Fig. 3A) and c.771_793del (p.Gly258Serfs*30) in exon 10 of CTNS gene in patient TN3 of family 2 (Fig. 3B) and one was previously unreported deletion of 20,327 bp in patient TN1 of family 1 (Fig. 3C).


Genetic basis of cystinosis in Tunisian patients: Identification of novel mutation in CTNS gene.

Chkioua L, Khedhiri S, Grissa O, Aloui C, Turkia HB, Ferchichi S, Miled A, Froissart R, Acquaviva C, Laradi S - Meta Gene (2015)

Direct sequencing of CTNS gene in Tunisian patients.A: Sequence electropherograms of exon 11 of the CTNS gene for c.1515G > A (p.G308R) mutation.B: Sequence electropherograms of exon 10 of the CTNS gene c.771_793del (p.Gly258Serfs*30) mutation.C: Characterization of the large rearrangement removing exons 1 to 5 of the CTNS gene and a part of the intron 1 of CARKL gene.1C: Schematic representation of the CTNS and CARKL genes rearrangement. Blue boxes represent SGSH exons and purple boxes represent CARLK exons. The 20,327 bp deleted region is delineated. The black arrows correspond to the primers (IVS1F_CARKL/IVS6R_CTNS) used to amplify the junction fragment.PCR of the deletion breakpoint. Lane 1: DNA 100-bp increment ladder; Lane 2: patient; Lane 3: patient; Lane 4: patient's mother; and Lane 5: control.The amplification showed a 540 bp product corresponding to the junction fragment in the patients and her mother. This product was not present in the control DNA sample.
© Copyright Policy - CC BY-NC-ND
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4528043&req=5

f0015: Direct sequencing of CTNS gene in Tunisian patients.A: Sequence electropherograms of exon 11 of the CTNS gene for c.1515G > A (p.G308R) mutation.B: Sequence electropherograms of exon 10 of the CTNS gene c.771_793del (p.Gly258Serfs*30) mutation.C: Characterization of the large rearrangement removing exons 1 to 5 of the CTNS gene and a part of the intron 1 of CARKL gene.1C: Schematic representation of the CTNS and CARKL genes rearrangement. Blue boxes represent SGSH exons and purple boxes represent CARLK exons. The 20,327 bp deleted region is delineated. The black arrows correspond to the primers (IVS1F_CARKL/IVS6R_CTNS) used to amplify the junction fragment.PCR of the deletion breakpoint. Lane 1: DNA 100-bp increment ladder; Lane 2: patient; Lane 3: patient; Lane 4: patient's mother; and Lane 5: control.The amplification showed a 540 bp product corresponding to the junction fragment in the patients and her mother. This product was not present in the control DNA sample.
Mentions: Analysis of the entire CTNS gene in all patients and their parents revealed the presence of three mutations (Table 3): Two of these mutations were previously described c.1515G > A (p.G308R) in exon 11 of CTNS gene in patient TN6 of family 3 (Fig. 3A) and c.771_793del (p.Gly258Serfs*30) in exon 10 of CTNS gene in patient TN3 of family 2 (Fig. 3B) and one was previously unreported deletion of 20,327 bp in patient TN1 of family 1 (Fig. 3C).

Bottom Line: Nephropathic cystinosis (NC) is an autosomal recessive disorder characterized by defective transport of cystine across the lysosomal membrane and resulting in renal, ophthalmic, and other organ abnormalities.In addition, eight polymorphisms were identified in the 3 patients and their parents.This study expands the mutational and population spectrum of NC, representing the first molecular diagnosis of NC in Tunisian population.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Biochemistry, F. Hached Hospital, 4000 Sousse, Tunisia ; University of Monastir, 5000 Monastir, Tunisia.

ABSTRACT
Nephropathic cystinosis (NC) is an autosomal recessive disorder characterized by defective transport of cystine across the lysosomal membrane and resulting in renal, ophthalmic, and other organ abnormalities. Mutations in the CTNS gene cause a deficiency of the transport protein, cystinosin. This study was performed to investigate mutations of the CTNS gene in three Tunisian families with NC. Polymerase chain reaction (PCR), ARMS multiplex PCR and direct sequencing were performed for molecular characterization of the CTNS gene in 3 unrelated Tunisian patients and their parents. Based on family history, prenatal diagnosis (PND) was performed in fetal DNA isolated from chorionic villi obtained at 10-12  weeks of gestation. None of the patients showed the most common 57-kb deletion in heterozygous or homozygous status. One patient was homozygous for the previously reported mutation c.1515G > A (p.G308R). One patient presented the novel gross deletion of 20,327 bp. One was homozygote for the previously reported mutation c.771_793del (p.Gly258Serfs*30). In addition, eight polymorphisms were identified in the 3 patients and their parents. The prenatal diagnosis in one family showed that the fetus DNA was heterozygous for the c.771_793del (p.Gly258Serfs*30) mutation. This study expands the mutational and population spectrum of NC, representing the first molecular diagnosis of NC in Tunisian population. The mutation screening of the CTNS gene was used for prenatal diagnosis to prevent and/or limit this inheritable disease in our country where the families are particularly large and have a high rate of consanguinity.

No MeSH data available.


Related in: MedlinePlus