Genetic analysis of essential cardiac transcription factors in 256 patients with non-syndromic congenital heart defects

Kazuki Kodo, Tsutomu Nishizawa, Michiko Furutani, Shoichi Arai, Kazuaki Ishihara, Mayumi Oda, Shinji Makino, Keiichi Fukuda, Takao Takahashi, Rumiko Matsuoka, Toshio Nakanishi, Hiroyuki Yamagishi

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

Background: The genetic basis of most congenital heart defects (CHDs), especially non-syndromic and non-familial conditions, remains largely unknown. Methods and Results: DNA samples were collected from immortalized cell lines and original genomes of 256 nonsyndromic, non-familial patients with cardiac outflow tract (OFT) defects. Genes encoding NKX2.5, GATA4, GATA6, MEF2C, and ISL1, essential for heart development, were analyzed using PCR-based bidirectional sequencing. The transcriptional activity of proteins with identified sequence variations was analyzed using a luciferase assay. A novel sequence variant (A103V in MEF2C) was identified, in addition to 4 unreported non-synonymous sequence variants in 3 known causative genes (A6V in NKX2.5, T330R and S339R in GATA4, and E142K in GATA6) in 5 individuals. None of these was found in 500 controls without CHDs. In vitro functional assay showed that all proteins with identified sequence variations exhibited significant changes in transcriptional activity and/or synergistic activity with other transcription factors. Furthermore, overexpression of the A103V MEF2C variant in a fish system disturbed early cardiac development. Conclusions: New mutations in the transcription factors NKX2.5, GATA4, GATA6, and MEF2C that affect their protein function were identified in 2.3% (6/256) of patients with OFT defects. Our results provide the first demonstration of MEF2C mutation and suggest that disturbances in the regulatory circuits involving these cardiac transcription factors may cause a subset of non-syndromic and non-familial CHDs.

Original languageEnglish
Pages (from-to)1703-1711
Number of pages9
JournalCirculation Journal
Volume76
Issue number7
DOIs
Publication statusPublished - 2012

Fingerprint

Congenital Heart Defects
Transcription Factors
GATA4 Transcription Factor
Mutation
Proteins
Luciferases
Genes
Fishes
Genome
Cell Line
Polymerase Chain Reaction
DNA

Keywords

  • Congenital heart defects
  • Genetics
  • Genotype
  • Pediatrics
  • Screening

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

Cite this

Genetic analysis of essential cardiac transcription factors in 256 patients with non-syndromic congenital heart defects. / Kodo, Kazuki; Nishizawa, Tsutomu; Furutani, Michiko; Arai, Shoichi; Ishihara, Kazuaki; Oda, Mayumi; Makino, Shinji; Fukuda, Keiichi; Takahashi, Takao; Matsuoka, Rumiko; Nakanishi, Toshio; Yamagishi, Hiroyuki.

In: Circulation Journal, Vol. 76, No. 7, 2012, p. 1703-1711.

Research output: Contribution to journalArticle

Kodo, Kazuki ; Nishizawa, Tsutomu ; Furutani, Michiko ; Arai, Shoichi ; Ishihara, Kazuaki ; Oda, Mayumi ; Makino, Shinji ; Fukuda, Keiichi ; Takahashi, Takao ; Matsuoka, Rumiko ; Nakanishi, Toshio ; Yamagishi, Hiroyuki. / Genetic analysis of essential cardiac transcription factors in 256 patients with non-syndromic congenital heart defects. In: Circulation Journal. 2012 ; Vol. 76, No. 7. pp. 1703-1711.
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abstract = "Background: The genetic basis of most congenital heart defects (CHDs), especially non-syndromic and non-familial conditions, remains largely unknown. Methods and Results: DNA samples were collected from immortalized cell lines and original genomes of 256 nonsyndromic, non-familial patients with cardiac outflow tract (OFT) defects. Genes encoding NKX2.5, GATA4, GATA6, MEF2C, and ISL1, essential for heart development, were analyzed using PCR-based bidirectional sequencing. The transcriptional activity of proteins with identified sequence variations was analyzed using a luciferase assay. A novel sequence variant (A103V in MEF2C) was identified, in addition to 4 unreported non-synonymous sequence variants in 3 known causative genes (A6V in NKX2.5, T330R and S339R in GATA4, and E142K in GATA6) in 5 individuals. None of these was found in 500 controls without CHDs. In vitro functional assay showed that all proteins with identified sequence variations exhibited significant changes in transcriptional activity and/or synergistic activity with other transcription factors. Furthermore, overexpression of the A103V MEF2C variant in a fish system disturbed early cardiac development. Conclusions: New mutations in the transcription factors NKX2.5, GATA4, GATA6, and MEF2C that affect their protein function were identified in 2.3{\%} (6/256) of patients with OFT defects. Our results provide the first demonstration of MEF2C mutation and suggest that disturbances in the regulatory circuits involving these cardiac transcription factors may cause a subset of non-syndromic and non-familial CHDs.",
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AU - Kodo, Kazuki

AU - Nishizawa, Tsutomu

AU - Furutani, Michiko

AU - Arai, Shoichi

AU - Ishihara, Kazuaki

AU - Oda, Mayumi

AU - Makino, Shinji

AU - Fukuda, Keiichi

AU - Takahashi, Takao

AU - Matsuoka, Rumiko

AU - Nakanishi, Toshio

AU - Yamagishi, Hiroyuki

PY - 2012

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N2 - Background: The genetic basis of most congenital heart defects (CHDs), especially non-syndromic and non-familial conditions, remains largely unknown. Methods and Results: DNA samples were collected from immortalized cell lines and original genomes of 256 nonsyndromic, non-familial patients with cardiac outflow tract (OFT) defects. Genes encoding NKX2.5, GATA4, GATA6, MEF2C, and ISL1, essential for heart development, were analyzed using PCR-based bidirectional sequencing. The transcriptional activity of proteins with identified sequence variations was analyzed using a luciferase assay. A novel sequence variant (A103V in MEF2C) was identified, in addition to 4 unreported non-synonymous sequence variants in 3 known causative genes (A6V in NKX2.5, T330R and S339R in GATA4, and E142K in GATA6) in 5 individuals. None of these was found in 500 controls without CHDs. In vitro functional assay showed that all proteins with identified sequence variations exhibited significant changes in transcriptional activity and/or synergistic activity with other transcription factors. Furthermore, overexpression of the A103V MEF2C variant in a fish system disturbed early cardiac development. Conclusions: New mutations in the transcription factors NKX2.5, GATA4, GATA6, and MEF2C that affect their protein function were identified in 2.3% (6/256) of patients with OFT defects. Our results provide the first demonstration of MEF2C mutation and suggest that disturbances in the regulatory circuits involving these cardiac transcription factors may cause a subset of non-syndromic and non-familial CHDs.

AB - Background: The genetic basis of most congenital heart defects (CHDs), especially non-syndromic and non-familial conditions, remains largely unknown. Methods and Results: DNA samples were collected from immortalized cell lines and original genomes of 256 nonsyndromic, non-familial patients with cardiac outflow tract (OFT) defects. Genes encoding NKX2.5, GATA4, GATA6, MEF2C, and ISL1, essential for heart development, were analyzed using PCR-based bidirectional sequencing. The transcriptional activity of proteins with identified sequence variations was analyzed using a luciferase assay. A novel sequence variant (A103V in MEF2C) was identified, in addition to 4 unreported non-synonymous sequence variants in 3 known causative genes (A6V in NKX2.5, T330R and S339R in GATA4, and E142K in GATA6) in 5 individuals. None of these was found in 500 controls without CHDs. In vitro functional assay showed that all proteins with identified sequence variations exhibited significant changes in transcriptional activity and/or synergistic activity with other transcription factors. Furthermore, overexpression of the A103V MEF2C variant in a fish system disturbed early cardiac development. Conclusions: New mutations in the transcription factors NKX2.5, GATA4, GATA6, and MEF2C that affect their protein function were identified in 2.3% (6/256) of patients with OFT defects. Our results provide the first demonstration of MEF2C mutation and suggest that disturbances in the regulatory circuits involving these cardiac transcription factors may cause a subset of non-syndromic and non-familial CHDs.

KW - Congenital heart defects

KW - Genetics

KW - Genotype

KW - Pediatrics

KW - Screening

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