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Targeted Next-Generation Sequencing Reveals Hot Spots and Doubly Heterozygous Mutations in Chinese Patients with Familial Cardiomyopathy.

Zhao Y, Feng Y, Zhang YM, Ding XX, Song YZ, Zhang AM, Liu L, Zhang H, Ding JH, Xia XS - Biomed Res Int (2015)

Bottom Line: Two hot spots (TNNI3-p.Arg145Gly, and LMNA-p.Arg190Trp) and double (LMNA-p.Arg190Trp plus MYH7-p.Arg1045His) heterozygous mutations were found to be highly correlated with familial cardiomyopathy.FDCM patients with doubly heterozygous mutations show a notably severe phenotype as we could confirm in our study; this indicates that the double mutations had a dose effect.In addition, it is proposed that genetic testing using NGS technology can be used as a cost-effective screening tool and help guide the treatment of patients with familial cardiomyopathy particularly regarding the risk of family members who are clinically asymptomatic.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Life Science and Technology, Center for Molecular Diagnosis in Yunnan Province, Kunming University of Science and Technology, Kunming 650500, China.

ABSTRACT
As a common cardiac disease mainly caused by gene mutations in sarcomeric cytoskeletal, calcium-handling, nuclear envelope, desmosomal, and transcription factor genes, inherited cardiomyopathy is becoming one of the major etiological factors of sudden cardiac death (SCD) and heart failure (HF). This disease is characterized by remarkable genetic heterogeneity, which makes it difficult to screen for pathogenic mutations using Sanger sequencing. In the present study, three probands, one with familial hypertrophic cardiomyopathy (FHCM) and two with familial dilated cardiomyopathy (FDCM), were recruited together with their respective family members. Using next-generation sequencing technology (NGS), 24 genes frequently known to be related to inherited cardiomyopathy were screened. Two hot spots (TNNI3-p.Arg145Gly, and LMNA-p.Arg190Trp) and double (LMNA-p.Arg190Trp plus MYH7-p.Arg1045His) heterozygous mutations were found to be highly correlated with familial cardiomyopathy. FDCM patients with doubly heterozygous mutations show a notably severe phenotype as we could confirm in our study; this indicates that the double mutations had a dose effect. In addition, it is proposed that genetic testing using NGS technology can be used as a cost-effective screening tool and help guide the treatment of patients with familial cardiomyopathy particularly regarding the risk of family members who are clinically asymptomatic.

No MeSH data available.


Related in: MedlinePlus

Results of the Sanger sequencing analysis. (a) shows the results for the TNNI3-p.Arg145Gly mutation in Family A; the results were positive in II: 3 and negative in III: 1; (b) shows the results for the LMNA-p.Arg190Trp mutation in Family B; the results were positive in II: 1, II: 3, and III: 1 and negative in I: 1; (c) shows the results for the MYH7-p.Arg1045His mutation in Family B; family members II: 1 and II: 3 tested positive, and III: 1 and I: 1 tested negative.
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fig3: Results of the Sanger sequencing analysis. (a) shows the results for the TNNI3-p.Arg145Gly mutation in Family A; the results were positive in II: 3 and negative in III: 1; (b) shows the results for the LMNA-p.Arg190Trp mutation in Family B; the results were positive in II: 1, II: 3, and III: 1 and negative in I: 1; (c) shows the results for the MYH7-p.Arg1045His mutation in Family B; family members II: 1 and II: 3 tested positive, and III: 1 and I: 1 tested negative.

Mentions: In family A, the proband (II: 3) was diagnosed with typical HCM. In the sequenced genomic regions, we identified the known C>T pathogenic heterozygotic mutation located at nucleotide position c.433 (Transcript name: NM_000363.4) in TNNI3 exon 7 (Table 1 and Figure 3(a)). This mutation results in the replacement of arginine at the 145th amino acid position by glycine (p.Arg145Gly). Using Sanger sequencing, this mutation was verified; it was not found in the daughter (Figure 3(a)). Further analysis showed that the amino acid Arg145 is highly conserved in many species and is localized in the first actin binding domain of the TNNI3 protein.


Targeted Next-Generation Sequencing Reveals Hot Spots and Doubly Heterozygous Mutations in Chinese Patients with Familial Cardiomyopathy.

Zhao Y, Feng Y, Zhang YM, Ding XX, Song YZ, Zhang AM, Liu L, Zhang H, Ding JH, Xia XS - Biomed Res Int (2015)

Results of the Sanger sequencing analysis. (a) shows the results for the TNNI3-p.Arg145Gly mutation in Family A; the results were positive in II: 3 and negative in III: 1; (b) shows the results for the LMNA-p.Arg190Trp mutation in Family B; the results were positive in II: 1, II: 3, and III: 1 and negative in I: 1; (c) shows the results for the MYH7-p.Arg1045His mutation in Family B; family members II: 1 and II: 3 tested positive, and III: 1 and I: 1 tested negative.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4495182&req=5

fig3: Results of the Sanger sequencing analysis. (a) shows the results for the TNNI3-p.Arg145Gly mutation in Family A; the results were positive in II: 3 and negative in III: 1; (b) shows the results for the LMNA-p.Arg190Trp mutation in Family B; the results were positive in II: 1, II: 3, and III: 1 and negative in I: 1; (c) shows the results for the MYH7-p.Arg1045His mutation in Family B; family members II: 1 and II: 3 tested positive, and III: 1 and I: 1 tested negative.
Mentions: In family A, the proband (II: 3) was diagnosed with typical HCM. In the sequenced genomic regions, we identified the known C>T pathogenic heterozygotic mutation located at nucleotide position c.433 (Transcript name: NM_000363.4) in TNNI3 exon 7 (Table 1 and Figure 3(a)). This mutation results in the replacement of arginine at the 145th amino acid position by glycine (p.Arg145Gly). Using Sanger sequencing, this mutation was verified; it was not found in the daughter (Figure 3(a)). Further analysis showed that the amino acid Arg145 is highly conserved in many species and is localized in the first actin binding domain of the TNNI3 protein.

Bottom Line: Two hot spots (TNNI3-p.Arg145Gly, and LMNA-p.Arg190Trp) and double (LMNA-p.Arg190Trp plus MYH7-p.Arg1045His) heterozygous mutations were found to be highly correlated with familial cardiomyopathy.FDCM patients with doubly heterozygous mutations show a notably severe phenotype as we could confirm in our study; this indicates that the double mutations had a dose effect.In addition, it is proposed that genetic testing using NGS technology can be used as a cost-effective screening tool and help guide the treatment of patients with familial cardiomyopathy particularly regarding the risk of family members who are clinically asymptomatic.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Life Science and Technology, Center for Molecular Diagnosis in Yunnan Province, Kunming University of Science and Technology, Kunming 650500, China.

ABSTRACT
As a common cardiac disease mainly caused by gene mutations in sarcomeric cytoskeletal, calcium-handling, nuclear envelope, desmosomal, and transcription factor genes, inherited cardiomyopathy is becoming one of the major etiological factors of sudden cardiac death (SCD) and heart failure (HF). This disease is characterized by remarkable genetic heterogeneity, which makes it difficult to screen for pathogenic mutations using Sanger sequencing. In the present study, three probands, one with familial hypertrophic cardiomyopathy (FHCM) and two with familial dilated cardiomyopathy (FDCM), were recruited together with their respective family members. Using next-generation sequencing technology (NGS), 24 genes frequently known to be related to inherited cardiomyopathy were screened. Two hot spots (TNNI3-p.Arg145Gly, and LMNA-p.Arg190Trp) and double (LMNA-p.Arg190Trp plus MYH7-p.Arg1045His) heterozygous mutations were found to be highly correlated with familial cardiomyopathy. FDCM patients with doubly heterozygous mutations show a notably severe phenotype as we could confirm in our study; this indicates that the double mutations had a dose effect. In addition, it is proposed that genetic testing using NGS technology can be used as a cost-effective screening tool and help guide the treatment of patients with familial cardiomyopathy particularly regarding the risk of family members who are clinically asymptomatic.

No MeSH data available.


Related in: MedlinePlus