title: Multiple gene variations contributed to congenital heart disease via GATA family transcriptional regulation.
abstract: Congenital heart disease (CHD) is a common birth defect, and most cases occur sporadically. Mutations in key genes that are responsible for cardiac development could contribute to CHD. To date, the genetic causes of CHD remain largely unknown.
  
  
  
  In this study, twenty-nine candidate genes in CHD were sequenced in 106 patients with Tetralogy of Fallot (TOF) using target exome sequencing (TES). The co-immunoprecipitation (CO-IP) and luciferase reporter gene assays were performed in HEK293T cells, and wild-type and mutant mRNA of ZFPM2 were microinjected into zebrafish embryos.
  
  
  
  Rare variants in key cardiac transcriptional factors and JAG1 were identified in the patients. Four patients carried multiple gene variants. The novel E1148K variant was located at the eighth Zinc-finger domain of FOG2 protein. The CO-IP assays in the HEK293T cells revealed that the variant significantly damaged the interaction between ZFPM2/FOG2 and GATA4. The luciferase reporter gene assays revealed that the E1148K mutant ZFPM2 protein displayed a significantly greater inhibition of the transcriptional activation of GATA4 than the wild-type protein. The wild-type mRNA and the E1148K mutant mRNA of ZFPM2 were injected into zebrafish embryos. At 48 hpf, in the mutant mRNA injection group, the number of embryos with an abnormal cardiac chamber structure and a loss of left-right asymmetry was increased. By 72 hpf, the defects in the chamber and left-right asymmetry became obvious.
  
  
  
  We performed TES in sporadic TOF patients and identified rare variants in candidate genes in CHD. We first validated the E1148 K variant in ZFPM2, which is likely involved in the pathogenesis of CHD via GATA4. Moreover, our results suggest that TES could be a useful tool for discovering sequence variants in CHD patients.
authors.affiliation: Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Collaborative Innovation Center of Genetics and Development, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai, 20032, China.|Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China.
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Qian
authors.affiliation: Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Collaborative Innovation Center of Genetics and Development, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai, 20032, China.
authors.forename: Deyong
authors.initials: D
authors.lastname: Xiao
authors.affiliation: Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Guo
authors.affiliation: Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China.
authors.forename: Hongbo
authors.initials: H
authors.lastname: Chen
authors.affiliation: Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Collaborative Innovation Center of Genetics and Development, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai, 20032, China.
authors.forename: Lili
authors.initials: L
authors.lastname: Hao
authors.affiliation: Pediatric Heart Center, Children's Hospital of Fudan University, Shanghai, 201102, China.
authors.forename: Xiaojing
authors.initials: X
authors.lastname: Ma
authors.affiliation: Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China.|Pediatric Heart Center, Children's Hospital of Fudan University, Shanghai, 201102, China.
authors.forename: Guoying
authors.initials: G
authors.lastname: Huang
authors.affiliation: Key Laboratory of Metabolism and Molecular Medicine, Ministry of Education, Department of Biochemistry and Molecular Biology, Collaborative Innovation Center of Genetics and Development, Institutes of Biomedical Sciences, School of Basic Medical Sciences, Fudan University, Shanghai, 20032, China. duanma@fudan.edu.cn.|Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China. duanma@fudan.edu.cn.|Research Center for Birth Defects, School of Basic Medical Sciences, Fudan University, 130 Dongan Road, Shanghai, 200030, China. duanma@fudan.edu.cn.
authors.forename: Duan
authors.initials: D
authors.lastname: Ma
authors.affiliation: Shanghai Key Lab of Birth Defect, Children's Hospital of Fudan University, Shanghai, 201102, China. huijunwang@fudan.edu.cn.|Molecular Genetics Laboratory, Children's Hospital of Fudan University, 399 Wanyuan Road, Shanghai, 201102, China. huijunwang@fudan.edu.cn.
authors.forename: Huijun
authors.initials: H
authors.lastname: Wang
mesh_terms: D000293:Adolescent; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002648:Child; D002675:Child, Preschool; D005260:Female; D050980:GATA Transcription Factors / Q000737:chemistry / Q000235:genetics*; D005786:Gene Expression Regulation; D014644:Genetic Variation; D057809:HEK293 Cells; D006330:Heart Defects, Congenital / Q000235:genetics*; D006801:Humans; D007223:Infant; D007231:Infant, Newborn; D008297:Male; D013771:Tetralogy of Fallot / Q000235:genetics; D014158:Transcription, Genetic; D015533:Transcriptional Activation / Q000235:genetics; D015027:Zebrafish
chemical_list: D050980:GATA Transcription Factors
keywords: Congenital heart disease (CHD); Functional study; Multigene disease; Next-generation sequencing; Variants