Heart formation, development, and differentiation in the human embryo must result from the ordered expression of specific gene products. Unfortunately, little is currently known about the genes and regulatory factors involved in cardiac morphogenesis in humans and other mammals. For this reason there is virtually no scientific foundation for understanding the causes of congenital heart disease. Our laboratory has recently identified a novel mouse transcription factor, termed GATA-4. This factor belongs to a group of related zinc finger proteins that recognize the consensus DNA sequence (a/T)GATA(A?G), known as the """"""""GATA"""""""" motif. Expression of GATA-4 is restricted to heart, with some expression also found in gonads, small intestinal epithelium, and yolk sac endoderm. Based on DNA-binding and co-transfection studies, we propose that GATA-4 functions as a transcriptional activator in heart and other tissues. The goal of this project is to elucidate the role of GATA-4 in cardiac gene expression and development. RNA analysis, in situ hybridization, and immunohistochemistry will be used to determine the pattern of GATA-4 expression in rodent heart during development and in response to hemodynamic overload and hormonal manipulation. The cis-elements regulating cardiac-specific expression of GATA-4 will be defined through deletional and mutational analysis of the promoter using several assay systems including transient transfection of cultured cardiocytes, stable transfection of pluripotential cell lines followed by differentiation into cardiac tissue, direct injection of rodent ventricle, and transgenic mice. To assess the role of GATA-4 in differentiation and development, targeted mutagenesis followed by gene conversion will be used to disrupt the GATA-4 gene in embryonic stem (ES) cells. The consequences of this mutation on in vitro differentiation of ES cell into myocardial and endodermal tissues will be determined. Chimeric mice will be derived by injecting blastocysts with ES cells containing a disrupted GATA-4 allele, and the subsequent breeding of these founder mice will produce GATA-4 homozygous deficient mice. Analysis of these mice will provide insight into the function of GATA-4 in the developing heart. Future studies will then build upon these results and provide a framework for detailed analyses of GATA-4 and its target genes in human hart during normal and pathological development and differentiation.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL052134-05
Application #
2702244
Study Section
Human Embryology and Development Subcommittee 1 (HED)
Project Start
1994-05-01
Project End
2000-04-30
Budget Start
1998-05-01
Budget End
2000-04-30
Support Year
5
Fiscal Year
1998
Total Cost
Indirect Cost
Name
Washington University
Department
Pediatrics
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
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Bielinska, Malgorzata; Seehra, Amrita; Toppari, Jorma et al. (2007) GATA-4 is required for sex steroidogenic cell development in the fetal mouse. Dev Dyn 236:203-13
Bielinska, Malgorzata; Jay, Patrick Y; Erlich, Jonathan M et al. (2007) Molecular genetics of congenital diaphragmatic defects. Ann Med 39:261-74
Heikinheimo, M; Ermolaeva, M; Bielinska, M et al. (1997) Expression and hormonal regulation of transcription factors GATA-4 and GATA-6 in the mouse ovary. Endocrinology 138:3505-14
Narita, N; Bielinska, M; Wilson, D B (1997) Cardiomyocyte differentiation by GATA-4-deficient embryonic stem cells. Development 124:3755-64
Narita, N; Bielinska, M; Wilson, D B (1997) Wild-type endoderm abrogates the ventral developmental defects associated with GATA-4 deficiency in the mouse. Dev Biol 189:270-4
Bielinska, M; Narita, N; Heikinheimo, M et al. (1996) Erythropoiesis and vasculogenesis in embryoid bodies lacking visceral yolk sac endoderm. Blood 88:3720-30
Bielinska, M; Wilson, D B (1995) Regulation of J6 gene expression by transcription factor GATA-4. Biochem J 307 ( Pt 1):183-9
White, R A; Dowler, L L; Pasztor, L M et al. (1995) Assignment of the transcription factor GATA4 gene to human chromosome 8 and mouse chromosome 14: Gata4 is a candidate gene for Ds (disorganization). Genomics 27:20-6
Ip, H S; Wilson, D B; Heikinheimo, M et al. (1994) The GATA-4 transcription factor transactivates the cardiac muscle-specific troponin C promoter-enhancer in nonmuscle cells. Mol Cell Biol 14:7517-26

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