Dominant-Negative ALK2 Allele Associates With Congenital Heart Defects

Background— Serious congenital heart defects occur as a result of improper atrioventricular septum (AVS) development during embryogenesis. Despite extensive knowledge of the genetic control of AVS development, few genetic lesions have been identified that are responsible for AVS-associated congenital heart defects. Methods and Results— We sequenced 32 genes known to be important in AVS development in patients with AVS defects and identified 11 novel coding single-nucleotide polymorphisms that are predicted to impair protein function. We focused on variants identified in the bone morphogenetic protein receptor, ALK2, and subjected 2 identified variants to functional analysis. The coding single-nucleotide polymorphisms R307L and L343P are heterozygous missense substitutions and were each identified in single individuals. The L343P allele had impaired functional activity as measured by in vitro kinase and bone morphogenetic protein–specific transcriptional response assays and dominant-interfering activity in vivo. In vivo analysis of zebrafish embryos injected with ALK2 L343P RNA revealed improper atrioventricular canal formation. Conclusion— These data identify the dominant-negative allele ALK2 L343P in a patient with AVS defects.

[1]  Jeroen Bakkers,et al.  Zebrafish integrin-linked kinase is required in skeletal muscles for strengthening the integrin-ECM adhesion complex. , 2008, Developmental biology.

[2]  P. Doevendans,et al.  Genes in congenital heart disease: atrioventricular valve formation , 2008, Basic Research in Cardiology.

[3]  Jeroen Bakkers,et al.  Rotation and asymmetric development of the zebrafish heart requires directed migration of cardiac progenitor cells. , 2008, Developmental cell.

[4]  Mauro W. Costa,et al.  Mutations in cardiac T-box factor gene TBX20 are associated with diverse cardiac pathologies, including defects of septation and valvulogenesis and cardiomyopathy. , 2007, American journal of human genetics.

[5]  A. Moorman,et al.  Role of bone morphogenetic proteins in cardiac differentiation. , 2007, Cardiovascular research.

[6]  C. Tabin,et al.  Bmp2 instructs cardiac progenitors to form the heart-valve-inducing field. , 2006, Developmental biology.

[7]  Edwin Cuppen,et al.  Generation of gene knockouts and mutant models in the laboratory rat by ENU-driven target-selected mutagenesis , 2006, Pharmacogenetics and genomics.

[8]  R. Schwartz,et al.  Bmp2 is essential for cardiac cushion epithelial-mesenchymal transition and myocardial patterning , 2005, Development.

[9]  H. Lehrach,et al.  Identification and functional analysis of CITED2 mutations in patients with congenital heart defects , 2005, Human mutation.

[10]  J. Epstein,et al.  Atrioventricular cushion transformation is mediated by ALK2 in the developing mouse heart. , 2005, Developmental biology.

[11]  B. Mulder,et al.  CONCOR, an initiative towards a national registry and DNA-bank of patients with congenital heart disease in the Netherlands: Rationale, design, and first results , 2005, European Journal of Epidemiology.

[12]  Joyce Bischoff,et al.  Heart valve development: endothelial cell signaling and differentiation. , 2004, Circulation research.

[13]  R. Schwartz,et al.  Bmp4 signaling is required for outflow-tract septation and branchial-arch artery remodeling. , 2004, Proceedings of the National Academy of Sciences of the United States of America.

[14]  Jonathan C. Cohen,et al.  GATA4 mutations cause human congenital heart defects and reveal an interaction with TBX5 , 2003, Nature.

[15]  C. Maslen,et al.  Missense mutations in CRELD1 are associated with cardiac atrioventricular septal defects. , 2003, American journal of human genetics.

[16]  C. Heldin,et al.  Elucidation of Smad Requirement in Transforming Growth Factor-β Type I Receptor-induced Responses* , 2003, The Journal of Biological Chemistry.

[17]  M. Goumans,et al.  Stimulation of Id1 Expression by Bone Morphogenetic Protein Is Sufficient and Necessary for Bone Morphogenetic Protein–Induced Activation of Endothelial Cells , 2002, Circulation.

[18]  J. Hoffman,et al.  The incidence of congenital heart disease. , 2002, Journal of the American College of Cardiology.

[19]  P. ten Dijke,et al.  Identification and Functional Characterization of Distinct Critically Important Bone Morphogenetic Protein-specific Response Elements in the Id1 Promoter* , 2002, The Journal of Biological Chemistry.

[20]  M. Mullins,et al.  Dorsoventral patterning in the zebrafish: bone morphogenetic proteins and beyond. , 2002, Results and problems in cell differentiation.

[21]  R. Kim,et al.  Bmp6 and Bmp7 are required for cushion formation and septation in the developing mouse heart. , 2001, Developmental biology.

[22]  J. Visser,et al.  The serine/threonine transmembrane receptor ALK2 mediates Müllerian inhibiting substance signaling. , 2001, Molecular endocrinology.

[23]  S. Henikoff,et al.  Predicting deleterious amino acid substitutions. , 2001, Genome research.

[24]  Warren C. Lathe,et al.  Prediction of deleterious human alleles. , 2001, Human molecular genetics.

[25]  Thomas N. Sato,et al.  Universal GFP reporter for the study of vascular development , 2000, Genesis.

[26]  M. Shaw,et al.  Activin receptor-like kinase 2 can mediate atrioventricular cushion transformation. , 2000, Developmental biology.

[27]  A. Bradley,et al.  Multiple roles for activin-like kinase-2 signaling during mouse embryogenesis. , 1999, Developmental biology.

[28]  P. Donahoe,et al.  The type I serine/threonine kinase receptor ActRIA (ALK2) is required for gastrulation of the mouse embryo. , 1999, Development.

[29]  R. Hennekam,et al.  Associations between chromosomal anomalies and congenital heart defects: a database search. , 1999, American journal of medical genetics.

[30]  P. Hoodless,et al.  Specific Activation of Smad1 Signaling Pathways by the BMP7 Type I Receptor, ALK2* , 1998, The Journal of Biological Chemistry.

[31]  J. Seidman,et al.  Congenital heart disease caused by mutations in the transcription factor NKX2-5. , 1998, Science.

[32]  L. Zon,et al.  The molecular nature of zebrafish swirl: BMP2 function is essential during early dorsoventral patterning. , 1997, Development.

[33]  C. Nüsslein-Volhard,et al.  Left-right pattern of cardiac BMP4 may drive asymmetry of the heart in zebrafish. , 1997, Development.

[34]  D A Kane,et al.  Genes establishing dorsoventral pattern formation in the zebrafish embryo: the ventral specifying genes. , 1996, Development.

[35]  M. Westerfield.  The zebrafish book : a guide for the laboratory use of zebrafish (Danio rerio) , 1995 .

[36]  K. Miyazono,et al.  Characterization of type I receptors for transforming growth factor-beta and activin. , 1994, Science.