Spectrum of heart disease associated with murine and human GATA4 mutation

doi:10.1016/j.yjmcc.2007.06.004

Journal of Molecular and Cellular Cardiology

Volume 43, Issue 6, December 2007, Pages 677-685

https://doi.org/10.1016/j.yjmcc.2007.06.004 Get rights and content

Abstract

The transcription factor GATA4 is essential for heart morphogenesis. Heterozygous mutation of GATA4 causes familial septal defects. However, the phenotypic spectrum of heterozygous GATA4 mutation is not known. In this study, we defined the cardiac phenotypes that result from heterozygous mutation of murine Gata4. We then asked if GATA4 mutation occurs in humans with these forms of congenital heart disease (CHD). In mice, heterozygous Gata4 mutation was associated with atrial and ventricular septal defect (ASD, VSD), endocardial cushion defect (ECD), RV hypoplasia, and cardiomyopathy. Genetic background strongly influenced the expression of ECD and cardiomyopathy, indicating the presence of important genetic modifiers. In humans, non-synonymous GATA4 sequence variants were associated with ECD (2/43), ASD (1/8), and RV hypoplasia in the context of double inlet left ventricle (1/9), forms of CHD that overlapped with abnormalities seen in the mouse model. These variants were not found in at least 500 control chromosomes, and encode proteins with non-conservative amino acid substitutions at phylogenetically conserved positions, suggesting that they are disease-causing mutations. Cardiomyopathy was not associated with GATA4 mutation in humans. These data establish the phenotypic spectrum of heterozygous Gata4 mutation in mice, and suggest that heterozygous GATA4 mutation leads to partially overlapping phenotypes in humans. Additional studies will be required to determine the degree to which GATA4 mutation contributes to human CHD characterized by ECD or RV hypoplasia.

Introduction

The morphogenetic complexity of fashioning a four-chambered heart from a straight tube mandates a precisely orchestrated interplay of multiple transcription factors, adhesion molecules, ion channels, signaling molecules and structural proteins [1]. Errors in this process result in congenital heart disease (CHD), the most common form of birth defect. Mutation of a small but growing number of genes has been shown to cause CHD [2]. Recently, mutation of the zinc finger transcription factor GATA4 was shown to cause atrial and ventricular septal defects in several unrelated extended pedigrees (Table 1) [3], [4], [5], [6]. Among CHD patients without a family history (“sporadic” CHD), GATA4 mutations appear to be infrequent. In published studies of sporadic CHD, GATA4 mutations were found in only 2 out of 376 probands examined (Table 1) [7], [8], [9], [10]. However, without prior knowledge of the phenotypic spectrum of GATA4 mutation, it was not possible to target these studies to forms of CHD most likely to be caused by GATA4 mutation. Therefore GATA4 mutation may be more frequently associated with specific forms of CHD that are not well represented in current literature.

We studied the spectrum of cardiac abnormalities found in mice with mutation of one copy of GATA4. We found that heterozygous GATA4 mutation in mice caused endocardial cushion defect (ECD), atrial or ventricular septal defect (ASD or VSD), hypoplastic right ventricle, and cardiomyopathy. Reasoning that this might give insight into the types of heart defects that might be caused by GATA4 mutation in humans, we then looked for GATA4 mutations in patients with similar forms of CHD. We found non-synonymous GATA4 sequence variants in association with ECD, hypoplastic RV in the context of double inlet left ventricle (DILV), and ASD. These results refine our understanding of the phenotypic spectrum of GATA4 mutation.

Section snippets

Mice

The Gata4^Δex2 allele has been described [11]. Mice were backcrossed for more than 7 generations into either the C57BL6/J (abbreviated C57; Jackson Labs) or the FVB/NCrl (FVB; Charles River Labs) genetic backgrounds. Structural abnormalities were diagnosed on H&E-stained serial paraffin sections. Echocardiography was performed on unsedated 8-week-old mice. Mice were held in a supine position and imaged with a 15-MHz probe. All analyses were performed blinded to genotype. Animal use was according

Murine model of Gata4 mutation

The mutant Gata4 allele used in this study, Gata4^Δex2, contains a deletion of the start codon and 46% of the coding region [11], [15]. This allele does not express full-length protein. In fetal heart, but not in adult heart, the allele does express a truncated protein lacking the essential N-terminal transcriptional activation domain (Fig. 1) [15]. In vitro reporter assays showed loss of transcriptional activity and did not suggest dominant negative activity [15]. Prior in vivo characterization

Spectrum of phenotypes associated with murine Gata4 mutation

Using tissue-restricted gene inactivation approaches, we previously showed that Gata4 is required in the myocardial compartment for normal myocardial growth and RV morphogenesis [24], and in endocardially derived structures for normal atrioventricular valve development [15]. These processes were also disrupted by a hypomorphic mutation of Gata4, which reduced Gata4 protein by 70% and resulted in embryonic lethality due to defects in myocardial growth, endocardial cushion development, and

Acknowledgments

This work was supported by grants from the NIH (SKR, NIH training grant T32HLO07572; WTP, SCCOR P50HLO74734; DWB SCCOR P50HL74728 and HL69712; and EG SCCOR P50HL74731). WTP was also supported by charitable donations from Edward Marran and Karen Carpenter. Jie Shen was supported as a Bang Bao Research Scholar by a grant from Proctor and Gamble. We thank M. Sarkar, C. Seidman, and J. Seidman for unpublished data.

References (32)

J. Ransom et al.
The genetics of cardiac birth defects
Semin. Cell Dev. Biol.
(2007)
W.T. Pu et al.
GATA4 is a dosage-sensitive regulator of cardiac morphogenesis
Dev. Biol.
(2004)
P.Y. Jay et al.
Impaired mesenchymal cell function in Gata4 mutant mice leads to diaphragmatic hernias and primary lung defects
Dev. Biol.
(2007)
D.B. McElhinney et al.
NKX2.5 mutations in patients with congenital heart disease
J. Am. Coll. Cardiol.
(2003)
E.E. Morrisey et al.
GATA-4 activates transcription via two novel domains that are conserved within the GATA-4/5/6 subfamily
J. Biol. Chem.
(1997)
S. Ghatpande et al.
Anterior endoderm is sufficient to rescue foregut apoptosis and heart tube morphogenesis in an embryo lacking retinoic acid
Dev. Biol.
(2000)
Y. Sakata et al.
The spectrum of cardiovascular anomalies in CHF1/Hey2 deficient mice reveals roles in endocardial cushion, myocardial and vascular maturation
J. Mol. Cell. Cardiol.
(2006)
D. Srivastava et al.
A genetic blueprint for cardiac development
Nature
(2000)
V. Garg et al.
GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5
Nature
(2003)
K. Hirayam-Yamada et al.
Phenotypes with GATA4 or NKX2.5 mutations in familial atrial septal defect
Am. J. Med. Genet., A
(2005)

A. Okubo et al.

A novel GATA4 mutation completely segregated with atrial septal defect in a large Japanese family

J. Med. Genet.

(2004)

A. Sarkozy et al.

Spectrum of atrial septal defects associated with mutations of NKX2.5 and GATA4 transcription factors

J. Med. Genet.

(2005)

A. Sarkozy et al.

CRELD1 and GATA4 gene analysis in patients with nonsyndromic atrioventricular canal defects

Am. J. Med. Genet., A

(2005)

G. Nemer et al.

A novel mutation in the GATA4 gene in patients with tetralogy of Fallot

Hum. Mutat.

(2006)

L. Zhang et al.

Screening of 99 Danish patients with congenital heart disease for GATA4 mutations

Genet. Test

(2006)

M.K. Schluterman et al.

Screening and biochemical analysis of GATA4 sequence variations identified in patients with congenital heart disease

Am. J. Med.Genet., A

(2007)

Cited by (211)

Y chromosome damage underlies testicular abnormalities in ATR-X syndrome
2024, iScience
ATR-X (alpha thalassemia, mental retardation, X-linked) syndrome features genital and testicular abnormalities including atypical genitalia and small testes with few seminiferous tubules. Our mouse model recapitulated the testicular defects when Atrx was deleted in Sertoli cells (ScAtrxKO) which displayed G2/M arrest and apoptosis. Here, we investigated the mechanisms underlying these defects. In control mice, Sertoli cells contain a single novel “GATA4 PML nuclear body (NB)” that contained the transcription factor GATA4, ATRX, DAXX, HP1α, and PH3 and co-localized with the Y chromosome short arm (Yp). ScAtrxKO mice contain single giant GATA4 PML-NBs with frequent DNA double-strand breaks (DSBs) in G2/M-arrested apoptotic Sertoli cells. HP1α and PH3 were absent from giant GATA4 PML-NBs indicating a failure in heterochromatin formation and chromosome condensation. Our data suggest that ATRX protects a Yp region from DNA damage, thereby preventing Sertoli cell death. We discuss Y chromosome damage/decondensation as a mechanism for testicular failure.
Genetics of Common Birth Defects in Newborns
2023, Principles of Neonatology
AbstractBirth defects are among the leading causes of morbidity and mortality in children and are present in 3% to 6% of births. The majority of birth defects are thought to be isolated and nonsyndromic at birth; however, as the child grows and develops, many are appreciated to be associated with other medical problems, difficulty with growth, and/or neurodevelopmental and behavioral issues. The etiologies for most birth defects are unknown and are likely multifactorial. However, as genomic technologies have matured and been used to interrogate large cohorts of individuals with birth defects, a range of genetic causes have been identified, including chromosome disorders, copy number variants, monogenic disorders, epigenetics, and common variants. In some cases, there may be contributions from both the maternal and fetal genomes because the mother’s genotype influences the metabolism of cofactors such as folate that may be critical to certain birth defects including neural tube defects. A limitation to the systematic analysis of the etiology of birth defects has been the limited availability of unbiased prospective data from mothers during pregnancy along with birth and long-term outcomes paired with comprehensive genomic data to assess the contribution of genes, the environment, and their interactions. Advances in genomic tools have now made it possible to genomically assess fetuses and newborns with birth defects to diagnose the 20% to 30% of cases with identifiable genetic etiologies and provide more accurate prognostic information and tailored surveillance as well as intervention to those infants likely to have associated medical and neurodevelopmental issues. In addition to supporting the care of the infant, this genetic information can provide important information to parents to accurately estimate the risk of recurrence and provide families with informed reproductive strategies for future pregnancies.
High-risk genes involved in common septal defects of congenital heart disease
2022, Gene
The septation defect is one of the main categories of congenital heart disease (CHD). They can affect the septation of the atria leading to atrial septal defect (ASD), septation of ventricles leading to ventricular septal defect (VSD), and formation of the central part of the heart leading to atrioventricular septal defect (AVSD). Disruption of critical genetic factors involved in the proper development of the heart structure leads to CHD manifestation. Because of this, to identify the high-risk genes involved in common septal defects, a comprehensive search of the literature with the help of databases and the WebGestalt analysis tool was performed. The high-risk genes identified in the analysis were checked in 16 Indian whole-exome sequenced samples, including 13 VSD and three Tetralogy of Fallot for in silico validation. This data revealed three variations in GATA4, i.e., c.C1223A at exon 6: c.C602A and c.C1220A at exon 7; and one variation in MYH6, i.e., c.G3883C at exon 28 in two VSD cases. This study supports previously published studies that suggested GATA4 and MYH6 as the high-risk genes responsible for septal defects. Thus, this study contributes to a better understanding of the genes involved in heart development by identifying the high-risk genes and interacting proteins in the pathway.
Transcription factor protein interactomes reveal genetic determinants in heart disease
2022, Cell
Congenital heart disease (CHD) is present in 1% of live births, yet identification of causal mutations remains challenging. We hypothesized that genetic determinants for CHDs may lie in the protein interactomes of transcription factors whose mutations cause CHDs. Defining the interactomes of two transcription factors haplo-insufficient in CHD, GATA4 and TBX5, within human cardiac progenitors, and integrating the results with nearly 9,000 exomes from proband-parent trios revealed an enrichment of de novo missense variants associated with CHD within the interactomes. Scoring variants of interactome members based on residue, gene, and proband features identified likely CHD-causing genes, including the epigenetic reader GLYR1. GLYR1 and GATA4 widely co-occupied and co-activated cardiac developmental genes, and the identified GLYR1 missense variant disrupted interaction with GATA4, impairing in vitro and in vivo function in mice. This integrative proteomic and genetic approach provides a framework for prioritizing and interrogating genetic variants in heart disease.
Rare variants in KDR, encoding VEGF Receptor 2, are associated with tetralogy of Fallot
2021, Genetics in Medicine
Rare genetic variants in KDR, encoding the vascular endothelial growth factor receptor 2 (VEGFR2), have been reported in patients with tetralogy of Fallot (TOF). However, their role in disease causality and pathogenesis remains unclear.
We conducted exome sequencing in a familial case of TOF and large-scale genetic studies, including burden testing, in >1,500 patients with TOF. We studied gene-targeted mice and conducted cell-based assays to explore the role of KDR genetic variation in the etiology of TOF.
Exome sequencing in a family with two siblings affected by TOF revealed biallelic missense variants in KDR. Studies in knock-in mice and in HEK 293T cells identified embryonic lethality for one variant when occurring in the homozygous state, and a significantly reduced VEGFR2 phosphorylation for both variants. Rare variant burden analysis conducted in a set of 1,569 patients of European descent with TOF identified a 46-fold enrichment of protein-truncating variants (PTVs) in TOF cases compared to controls (P = 7 × 10^-11).
Rare KDR variants, in particular PTVs, strongly associate with TOF, likely in the setting of different inheritance patterns. Supported by genetic and in vivo and in vitro functional analysis, we propose loss-of-function of VEGFR2 as one of the mechanisms involved in the pathogenesis of TOF.
Rs4841587 in GATA4 and rs6999593 in DNMT1 gene associated with congenital heart diseases in the southeast of Iran
2020, Meta Gene
Different genetic and epigenetic variations are implicated in congenital heart diseases (CHDs). GATA4 and DNMT1 genes play an important role in embryonic heart development. We aim to investigate the correlation between rs4841587 in GATA4 and rs6999593 in DNMT1 genes with susceptibility and severity of symptoms in ventricular septal defects (VSDs) and tetralogy of fallot (TF) diseases in the southeast of Iran.
This case-control study was evaluated 200 children diagnosed with VSDs or TF as well as 200 healthy controls. SNPs genotyping were performed using T-ARMS-PCR. Odds ratio (OR) and 95% confidence interval (CI), as well as Chi-square, was used to assess the strength of the association.
TT genotype at rs4841587 in GATA4 was associated with the susceptibility of CHDs (OR = 2.02, 95% CI = 1.06–3.83, P = .031). Rs4841587 was related with VSDs under the dominant model (GT + TT vs. GG: OR = 1.70, 95% CI = 1.08–2.65, P = .020). However, no significant differences were observed in genotype frequencies between TF cases and controls at this position. AG genotype at rs6999593 in DNMT1 was strongly correlated with the risk of CHDs (OR = 2.27, 95% CI = 1.43–3.60, P = .000). Significant differences also were found between AG genotype with VSDs (OR = 2.12, 95% CI = 1.19–3.77, P = .011) and TF (OR = 2.58, 95% CI = 1.19–5.57, P = .016).
rs4841587 and rs6999593 can be proposed as a significant marker for CHDs. Rs4841587 T/G polymorphism correlated with the susceptibility and severity of symptoms in VSDs. Moreover, rs6999593 A/G affects the susceptibility of VSDs and TF, in the southeast of Iran. However, further investigations are necessary to confirm these results.

View all citing articles on Scopus

¹: Current address: Cardiology Department, Shanghai Children’s Hospital, Shanghai Jiaotong University, 24 Lane 1400 Western Beijing Road, Shanghai 200040, China.

²: Current address: Department of Surgery, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53226, USA.

³: Current address: Gladstone Institute of Cardiovascular Disease, 1650 Owens Street, San Francisco, CA 94158, USA.

View full text

Original articleSpectrum of heart disease associated with murine and human GATA4 mutation

Abstract

Introduction

Section snippets

Mice

Murine model of Gata4 mutation

Spectrum of phenotypes associated with murine Gata4 mutation

Acknowledgments

Semin. Cell Dev. Biol.

Dev. Biol.

Dev. Biol.

J. Am. Coll. Cardiol.

J. Biol. Chem.

Dev. Biol.

J. Mol. Cell. Cardiol.

A genetic blueprint for cardiac development

Nature

GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5

Nature

Phenotypes with GATA4 or NKX2.5 mutations in familial atrial septal defect

Am. J. Med. Genet., A

A novel GATA4 mutation completely segregated with atrial septal defect in a large Japanese family

J. Med. Genet.

Spectrum of atrial septal defects associated with mutations of NKX2.5 and GATA4 transcription factors

J. Med. Genet.

CRELD1 and GATA4 gene analysis in patients with nonsyndromic atrioventricular canal defects

Am. J. Med. Genet., A

A novel mutation in the GATA4 gene in patients with tetralogy of Fallot

Hum. Mutat.

Screening of 99 Danish patients with congenital heart disease for GATA4 mutations

Genet. Test

Screening and biochemical analysis of GATA4 sequence variations identified in patients with congenital heart disease

Am. J. Med.Genet., A

Original article
Spectrum of heart disease associated with murine and human GATA4 mutation