Vesa M Kaartinen
University Of Michigan At Ann Arbor
Project start date: 2001-02-01
Project end date: 2015-02-28
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THE ROLE OF TGF-BETA3 IN PALATOGENESIS
Vesa M Kaartinen, Assistant Professor
Biologic And Materials Sciencesuniversity Of Michigan At Ann Arbor
Grant 5R01DE013085-10 from National Institute Of Dental & Craniofacial Research IRG: SBDD
Abstract: The long-term goal of this proposal is to understand specific molecular mechanisms of Tgf-beta signaling that take place during palatal fusion. This is very important since failure of these processes has been shown to lead to cleft palate, one of the most common birth defects in humans. Others and we have previously shown that during palatal fusion transforming growth factor-betas (Tgf-betas) induce several downstream responses that contribute to the successful palatogenesis. This proposal focuses on the role of epithelial and mesenchymal Tgf-beta signals in coordination of palatal fusion. Our overall hypothesis is Tgf-beta/Bmp signaling events both in the palatal mesenchyme and in the epithelium are critical for successful palatal fusion and play a concerted role during palatogenesis. We propose to test this hypothesis by five Specific Aims. In Aim 1 we propose to determine the effect of neural crest cell specific abrogation of the Tgf-b type I receptor AIk2 on palatal development. In Aim 2, we propose to define the role of Tgf-beta type I receptors AIk2 and AIk5 in the palatal midline epithelium during palatal fusion, in Aim 3 we propose to determine the role of reciprocal epithelio-mesenchymal Tgf-beta signaling during palatal fusion, in Aim 4 we propose to analyze why direct contact between the palatal epithelium and mesenchyme can bypass the Tgf-beta3 signaling defect, and finally in Aim 5 we propose to determine the role of gap junctions in Tgf-beta-induced palatal midline epithelial fusion. Our unique experimental models and the state-of-art strategy allow us to determine the role Tgf-beta signaling via AIk5 and AIk2 in palatogenesis. Collectively, the proposed experiments are likely to be of critical importance in attempting to understand the molecular bases of facial and palatal cleftings in humans, and to develop possible therapeutic approaches to treat cleft palate during the fetal period
Keywords: biological signal transduction, bone morphogenetic protein, cellular pathology, cleft palate, growth factor receptor, molecular pathology, transforming growth factor cell cell interaction, cell migration, epithelium, gap junction, gene expression, genetic promoter element, membrane channel, mesenchyme, mutant, neural crest, protein structure function genetically modified animal, laboratory mouse, tissue /cell culture
Project start date: 2001-02-01
Project end date: 2009-12-31
5R01DE013085-07 (2007): $361020
5R01DE013085-06 (2006): $371802
5R01DE013085-04 (2004): $260925
5R01DE013085-03 (2003): $260925
5R01DE013085-02 (2002): $260925
Grants awarded to Vesa M Kaartinen
THE ROLE OF TGF-BETA3 IN PALATOGENESIS
Vesa M Kaartinen, Assistant Professor
Children s Hospital Los Angeles Los Angeles, Ca 900276062
Grant 2R01DE013085-05 from National Institute Of Dental And Craniofacial Research IRG: SBDD
Abstract: The long-term goal of this proposal is to understand specific molecular mechanisms of Tgf-beta signaling that take place during palatal fusion. This is very important since failure of these processes has been shown to lead to cleft palate, one of the most common birth defects in humans. Others and we have previously shown that during palatal fusion transforming growth factor-betas (Tgf-betas) induce several downstream responses that contribute to the successful palatogenesis. This proposal focuses on the role of epithelial and mesenchymal Tgf-beta signals in coordination of palatal fusion. Our overall hypothesis is Tgf-beta/Bmp signaling events both in the palatal mesenchyme and in the epithelium are critical for successful palatal fusion and play a concerted role during palatogenesis. We propose to test this hypothesis by five Specific Aims. In Aim 1 we propose to determine the effect of neural crest cell specific abrogation of the Tgf-b type I receptor AIk2 on palatal development. In Aim 2, we propose to define the role of Tgf-beta type I receptors AIk2 and AIk5 in the palatal midline epithelium during palatal fusion, in Aim 3 we propose to determine the role of reciprocal epithelio-mesenchymal Tgf-beta signaling during palatal fusion, in Aim 4 we propose to analyze why direct contact between the palatal epithelium and mesenchyme can bypass the Tgf-beta3 signaling defect, and finally in Aim 5 we propose to determine the role of gap junctions in Tgf-beta-induced palatal midline epithelial fusion. Our unique experimental models and the state-of-art strategy allow us to determine the role Tgf-beta signaling via AIk5 and AIk2 in palatogenesis. Collectively, the proposed experiments are likely to be of critical importance in attempting to understand the molecular bases of facial and palatal cleftings in humans, and to develop possible therapeutic approaches to treat cleft palate during the fetal period.
Keywords: biological signal transduction, bone morphogenetic protein, cellular pathology, cleft palate, growth factor receptor, molecular pathology, transforming growth factor, cell cell interaction, cell migration, epithelium, gap junction, gene expression, genetic promoter element, membrane channel, mesenchyme, mutant, neural crest, protein structure function, genetically modified animal, laboratory mouse, tissue /cell culture
Project start date: 2001-02-01
Project end date: 2009-12-31
2R01DE013085-05 (2005): $380750
1R01DE013085-01A2 (2001): $260925
Activin Receptor-Like Kinase-2 In Cardiac Morphogenesis
Vesa M Kaartinen, Assistant Professor
Children s Hospital Los Angeles Los Angeles, Ca 900276062
Grant 5R01HL074862-04 from National Heart, Lung, And Blood Institute IRG: HED
Abstract: Malformations of the cardiovascular system are among the most common birth defects in humans. Members of the evolutionary conserved TGF-beta superfamily of secreted growth factors have been shown to play important roles during normal heart development. They all signal through several different type I receptors (called Activin Receptor-Like Kinases; Alks), which are the primary determinants of signaling specificity. Among these receptors AIk2 is of particular interest, since it appears that AIk2 mediates both BMP, Activin and TGFbeta signals. AIk2 is strongly expressed in the heart, and our preliminary experiments demonstrate that AIk2 is required for normal cardiac development. Therefore, we hypothesize that AIk2 plays a key role in cardiac outflow tract development by regulating differentiation, proliferation and/or survival of cardiac neural crest cells. We will test this hypothesis by three Specific Aims by utilizing a genetically manipulated mouse strain we recently developed that allows inactivation of AIk2 specifically in neural crest cells. In Aim 1, defects in valves, septa and outflow tracts will be analyzed in detail. In Aim 2, we will define the specific process controlled by AIk2 during cardiac outflow tract morphogenesis, and in Aim 3 we will identify the relevant downstream signaling molecules, Smads, and transcriptional targets in the AIk2-mediated pathway. Our experimental strategy will allow us to determine the biological role of AIk2 in cardiac outflow tract development. The results of these studies are likely to be of critical importance in attempting to reach our long-term goal to understand the molecular basis of life-threatening congenital heart defects in humans.
Keywords: cardiogenesis, congenital heart disorder, growth factor receptor, protein structure function, activin, biological signal transduction, bone morphogenetic protein, cell differentiation, cell growth regulation, cell proliferation, genetic transcription, morphology, neural crest, transforming growth factor, genetically modified animal, laboratory mouse, microarray technology
Project start date: 2004-02-01
Project end date: 2009-01-31
5R01HL074862-04 (2007): $264742
5R01HL074862-03 (2006): $290606
5R01HL074862-02 (2005): $297600
1R01HL074862-01 (2004): $292720
5R01HL074862-07 (2010): $373320
5R01HL074862-06 (2009): $373320
THE ROLE OF TGF-BETA3 IN PALATOGENESIS
Vesa M Kaartinen, Associate Professor
University Of Michigan At Ann Arbor, 1040 Wolverine Tower, Ann Arbor, Mi 48109-1274
Grant 2R01DE013085-11 from National Institute Of Dental & Craniofacial Research
Abstract: Cleft palate is among the most common birth defects in humans. We and others have shown that transforming growth factor-¿3 (Tgf-¿3) plays a critical role in palatal epithelial fusion both in mice and in humans. Failure in Tgf-¿3 signaling leads to a condition in which the palatal medial edge epithelium fails to disappear, and subsequently fully- grown palatal shelves fail to fuse. During previous funding years of this grant, we have shown that Tgf-¿ signaling is required both in the palatal epithelium and in the palatal mesenchyme for appropriate palatal shelf growth, patterning and fusion. Consequently, our results suggest that the role of Tgf-¿3 during palatogenesis is broader and more complex than previously thought. Based on the recent findings and our preliminary data, we have formulated an overall hypothesis that specific, modular, long-distance regulatory elements are responsible of targeting Tgf¿3 expression to the prefusion palatal epithelium. This epithelial-specific expression of Tgf¿3, in turn, induces both Smad4-dependent (canonical) and Smad4-independent (non-canonical) signaling events in the palatal epithelium that are together with the mesenchymal Tgf-¿3 signaling, required for successful palatal fusion. In specific aim 1, we propose to test the hypothesis that Tgf¿3 transcription in palatal medial edge epithelial cells is regulated by distant 5´ and/or 3´ enhancers. In specific aim 2, we propose to test the hypothesis that Tgf-¿-activated kinase-1 (Tak1) functions together with the Smad4-mediated signaling pathway to induce Tgf-¿3-induced cell cycle arrest and subsequent apoptosis in palatal epithelial cells. In specific aim 3, we propose to test the hypothesis that in Tgfb3 null mutants, imbalance between Tgf-¿ and Notch signaling leads to an attenuated expression of cell cycle inhibitors ultimately resulting in cleft palate, and finally, in specific aim 4, we propose to test the hypothesis that Tgf-¿3 plays a previously unexpected role in the palatal mesenchyme by regulating appropriate patterning of prefusion palatal shelves. Our unique experimental models and state-of-art strategy will allow us to define to the role of canonical and non-canonical Tgf-¿ signaling during palatogenesis. Collectively, the proposed experiments are likely to be of critical importance in attempting to understand the molecular basis of the cleft palate syndrome in humans. Cleft palate is one of the most common congenital birth defects in humans. The proposed studies dissect the role of a transforming growth factor-beta signaling in the pathogenesis of cleft palate. We expect that the proposed studies will be important in understanding of the underlying molecular mechanisms that lead to the cleft palate syndrome in humans
Keywords: Animal growth regulators, transforming growth factors; Apoptosis; Apoptosis Pathway; Arts; Attenuated; BACs (Chromosomes); Bacterial Artificial Chromosomes; Birth Defects; Body Tissues; Bone-Derived Transforming Growth Factor; CSBP1; CSBP2; CSPB1; Cell Communication and Signaling; Cell Cycle; Cell Cycle Arrest; Cell Death, Programmed; Cell Division Cycle; Cell Growth in Number; Cell Multiplication; Cell Proliferation; Cell Signaling; Cellular Proliferation; Cleft Palate; Complex; Congenital Abnormality; Congenital Anatomic Abnormality; Congenital Anatomical Abnormality; Congenital Defects; Congenital Deformity; Congenital Malformation; Culturing, in vitro Organ; Culturing, in vitro Vertebrate, Organ; Data; Distant; EC 2.7; EXIP; Engineering; Engineerings; Enhancers; Epithelial; Epithelial Cells; Epithelium; Event; Experimental Models; Experimental Models, Other; FLR; Failure (biologic function); Funding; Gene Expression; Gene Transcription; Generalized Growth; Genes; Genes, LacZ; Genetic Transcription; Genomics; Grant; Growth; Health; Human; Human, General; In Vitro; Intracellular Communication and Signaling; Kinases; LacZ; LacZ Genes; Lead; MAPK14; MAPK14 gene; Mammals, Mice; Man (Taxonomy); Man, Modern; Medial; Mediating; Mesenchymal; Mesenchymas; Mesenchyme; Methods and Techniques; Methods, Other; Mice; Mice, Transgenic; Milk Growth Factor; Models, Experimental; Molecular; Molecular Genetic Abnormality; Morphology; Murine; Mus; Mxi2; Organ Culture; Organ Culture Techniques; PRKM14; PRKM15; Pathogenesis; Pattern; Pb element; Phosphotransferases; Platelet Transforming Growth Factor; Play; RNA Expression; Regulatory Element; RegulatoryElement; Reporter; Reporting; Role; SAPK2A; Signal Pathway; Signal Transduction; Signal Transduction Systems; Signaling; Surface; Survey Instrument; Surveys; Syndrome; TGF B; TGF-Beta-3; TGF-beta; TGF-beta3; TGFB3; TGFbeta; Techniques; Testing; Tissue Growth; Tissues; Transcription; Transcription, Genetic; Transforming Growth Factor Beta 3; Transforming Growth Factor beta; Transforming Growth Factors; Transgenic Mice; Transgenic Organisms; Transphosphorylases; Tumor Growth Factors; Work; base; biological signal transduction; experiment; experimental research; experimental study; failure; heavy metal Pb; heavy metal lead; in vivo; inhibitor; inhibitor/antagonist; knockout gene; mutant; notch; notch protein; notch receptors; novel; ontogeny; p38; p38 MAPK Gene; p38Alpha; palatal fusion; palatal shelves; palatogenesis; public health relevance; research study; social role; transforming growth factor beta3; transgenic
Relevance: Cleft palate is one of the most common congenital birth defects in humans. The proposed studies dissect the role of a transforming growth factor-beta signaling in the pathogenesis of cleft palate. We expect that the proposed studies will be important in understanding of the underlying molecular mechanisms that lead to the cleft palate syndrome in humans
Project start date: 2001-02-01
Project end date: 2015-02-28
Budget start date: 15-MAR-2010
Budget end date: 28-FEB-2011
PFA/PA: PA-07-070
2R01DE013085-11 (2010): $358803