title: Gene network and familial analyses uncover a gene network involving Tbx5/Osr1/Pcsk6 interaction in the second heart field for atrial septation.
abstract: Atrial septal defects (ASDs) are a common human congenital heart disease (CHD) that can be induced by genetic abnormalities. Our previous studies have demonstrated a genetic interaction between Tbx5 and Osr1 in the second heart field (SHF) for atrial septation. We hypothesized that Osr1 and Tbx5 share a common signaling networking and downstream targets for atrial septation. To identify this molecular networks, we acquired the RNA-Seq transcriptome data from the posterior SHF of wild-type, Tbx5(+/) (-), Osr1(+/-), Osr1(-/-) and Tbx5(+/-)/Osr1(+/-) mutant embryos. Gene set analysis was used to identify the Kyoto Encyclopedia of Genes and Genomes pathways that were affected by the doses of Tbx5 and Osr1. A gene network module involving Tbx5 and Osr1 was identified using a non-parametric distance metric, distance correlation. A subset of 10 core genes and gene-gene interactions in the network module were validated by gene expression alterations in posterior second heart field (pSHF) of Tbx5 and Osr1 transgenic mouse embryos, a time-course gene expression change during P19CL6 cell differentiation. Pcsk6 was one of the network module genes that were linked to Tbx5. We validated the direct regulation of Tbx5 on Pcsk6 using immunohistochemical staining of pSHF, ChIP-quantitative polymerase chain reaction and luciferase reporter assay. Importantly, we identified Pcsk6 as a novel gene associated with ASD via a human genotyping study of an ASD family. In summary, our study implicated a gene network involving Tbx5, Osr1 and Pcsk6 interaction in SHF for atrial septation, providing a molecular framework for understanding the role of Tbx5 in CHD ontogeny.
authors.affiliation: Department of Pathology, School of Medicine and Health Sciences, ND INBRE Bioinformatics Core, University of North Dakota, Grand Forks, ND 58202, USA.
authors.forename: Ke K
authors.initials: KK
authors.lastname: Zhang
authors.affiliation: Department of Basic Sciences, School of Medicine and Health Sciences and ND INBRE Bioinformatics Core, University of North Dakota, Grand Forks, ND 58202, USA.
authors.forename: Menglan
authors.initials: M
authors.lastname: Xiang
authors.affiliation: Department of Basic Sciences, School of Medicine and Health Sciences and Department of Gerontology, Tongji Hospital, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
authors.forename: Lun
authors.initials: L
authors.lastname: Zhou
authors.affiliation: Department of Basic Sciences, School of Medicine and Health Sciences and.
authors.forename: Jielin
authors.initials: J
authors.lastname: Liu
authors.affiliation: Department of Basic Sciences, School of Medicine and Health Sciences and.
authors.forename: Nathan
authors.initials: N
authors.lastname: Curry
authors.affiliation: Laboratori de Genetica Molecular, Hospital Son Espases, Palma de Mallorca 07010, Spain.
authors.forename: Damian
authors.initials: D
authors.lastname: Heine Suñer
authors.affiliation: Department of Cardiology, Heart Failure and Inherited Cardiac Diseases Unit, Hospital Universitario Puerta de Hierro Majadahonda, Manuel de Falla, 1, 28222 Majadahonda, Madrid, Spain.
authors.forename: Pablo
authors.initials: P
authors.lastname: Garcia-Pavia
authors.affiliation: Nemours Research Institute, Nemours Children's hospital, Orlando, FL 32827, USA.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Zhang
authors.affiliation: Department of Molecular Cardiology, Center for Cardiovascular Genetics, Lerner Research Institute, Cleveland Clinic, Cleveland, OH 44195, USA, Department of Molecular Medicine and Department of Genetics and Genome Sciences, Case Western Reserve University, Cleveland, OH 44106, USA and.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wang
authors.affiliation: Department of Basic Sciences, School of Medicine and Health Sciences and Department of Nutrition and Food Science, Texas A&M University, Cater-Mattil Hall Rm 217B, TAMU 2253, College Station, TX 77843, USA linglin.xie@tamu.edu.
authors.forename: Linglin
authors.initials: L
authors.lastname: Xie
mesh_terms: D000818:Animals; D053263:Gene Regulatory Networks; D057809:HEK293 Cells; D006330:Heart Defects, Congenital / Q000235:genetics / Q000378:metabolism; D006344:Heart Septal Defects, Atrial / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D010375:Pedigree; D043484:Proprotein Convertases / Q000235:genetics; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics* / Q000378:metabolism; D015398:Signal Transduction; D020825:T-Box Domain Proteins / Q000235:genetics* / Q000378:metabolism
chemical_list: D020825:T-Box Domain Proteins; C103172:T-box transcription factor 5; C118135:OXSR1 protein, human; C534421:OXSR1 protein, mouse; D017346:Protein Serine-Threonine Kinases; C080654:PCSK6 protein, human; D043484:Proprotein Convertases; D012697:Serine Endopeptidases