Regulation of EMT During Avian Heart Valve Formation

禽类心脏瓣膜形成过程中 EMT 的调节

• 批准号: 7267533
• 负责人: RAYMOND Bruce RUNYAN
• 金额: $ 36.68万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7267533
• 关键词: ACVR1 gene; Adult; Antibodies; Antisense Oligonucleotides; Biological Assay; Biology; Birds; Cardiac; Cell Death; Cell Separation; Cell physiology; Cells; Collagen; Complex; Cytoskeleton; Data; Development; Dimensions; Disruption; Dissociation; Elements; Embryo; Embryonic Heart; Endoglin; Endopeptidases; Endothelial Cells; Epithelial; Epithelium; Extracellular Matrix; Family; Fibrosis; Gel; Gene Expression; Genes; Growth Factor; Heart; Heart Valves; Homeobox; Hyaluronan; Invasive; Islets of Langerhans; Kidney; Laboratories; Laboratory Study; Lateral; Ligands; Light; Mediating; Mediator of activation protein; Mesenchymal; Modeling; Mus; Neoplasm Metastasis; Neural Crest; Neural Crest Cell; Neural tube; Neuregulins; Outcome; Pathologic Processes; Peptide Hydrolases; Phenotype; Polymerase Chain Reaction; Population; Process; Production; Protein Isoforms; Proteins; Regulation; Role; Shapes; Signal Transduction; Site; Somites; Staging; Surface; System; TGF-beta type I receptor; TGFB1 gene; TGFB3 gene; Time; Tissues; Transforming Growth Factor Beta 2; Tricuspid valve structure; Variant; Vascular Endothelial Growth Factors; Vertebrates; Work; Wound Healing; Zebrafish; Zinc Fingers; cadherin 5; cardiogenesis; cell motility; cell transformation; cell type; congenital heart disorder; experimental analysis; migration; novel; progenitor; receptor; receptor expression; research study; response; slug; transcription factor; two-dimensional

DESCRIPTION (provided by applicant): During cardiac development, valve progenitors first arise in the atrioventricular canal as a result of a TGF¿-mediated epithelial-mesenchymal cell transformation (EMT). Prior studies showed that this EMT can be divided into two stages, an activation stage and an invasion stage. Stage 1 is mediated, in part, by transforming growth factor beta 2 (TGF¿2) and stage 2 is mediated, in part, by TGF¿3. Previous work showed that EMT can be disrupted by the inhibition of 5 different TGF¿ receptors, TGF¿ receptor II, TGF¿ receptor III, Endoglin, ALK2 and ALK5. It is our hypothesis that there are two more discrete signaling complexes mediating TGF¿-regulated EMT. To continue the analysis of TGF¿-mediated regulation of normal heart development, the first aim will examine the correlation between specific TGF¿ isoforms and receptors in mediating EMT regulation. Microarray experiments will be performed to assess the effect loss of specific TGF¿ isoforms on altered gene expression in tissues prior to EMT, during EMT and in mesenchymal cells after EMT. Selected markers among the TGF¿-regulated genes will then be used to assess whether gene expression is altered as each of the specific receptors is disrupted. Correlations between these data will point to specific receptor and ligand signaling complexes that mediate EMT. These putative complexes will be confirmed at the protein level. Data from other labs has implicated a variety of signal transduction mechanisms and intracellular regulators in cardiac EMT including ErbB2, NF-1, VEGF and NFATc1. In the second aim, experiments will be undertaken to determine whether disruption of TGF¿ signaling alters any of the other candidate mechanisms or whether loss of these molecules alters TGF¿ ligand or receptor expression. It was observed in the neural crest system that forced expression of Sox 8, 9 or 10 in the neural tube would cause an EMT and produce ectopic neural crest cells. We conjecture that this is analogous to the cell invasion step in the heart and that it requires some TGF¿ activity. In the third aim, experiments will be undertaken to determine whether exogenous expression of Sox genes will force EMT and whether Sox gene expression is regulated by TGF¿ signal transduction or regulates TGF¿-mediated responses. Together these experiments will shed new light on the mechanisms of EMT regulation and aid in an understanding of congenital heart disease.

描述（由申请人提供）：在心脏发育过程中，由于 TGF 的作用，瓣膜祖细胞首先在房室管中出现。介导的上皮间质细胞转化 (EMT) 先前的研究表明，这种 EMT 可以分为两个阶段，即激活阶段和侵袭阶段，第 1 阶段部分是由转化生长因子 β 2 (TGF¿2) 介导的。 ），第 2 阶段部分由 TGF¿ 3. 之前的研究表明，EMT 可以通过抑制 5 种不同的 TGF 来破坏？受体、TGF¿受体 II、TGF¿受体 III、内皮糖蛋白、ALK2 和 ALK5 我们假设有两种更离散的信号复合物介导 TGF¿ -调节EMT，继续分析TGF¿ -介导正常心脏发育的调节，第一个目标将检查特定 TGF 之间的相关性将进行介导 EMT 调节的同工型和受体的微阵列实验，以评估特定 TGF 的效应损失。 EMT 之前、EMT 期间和 EMT 后间充质细胞中基因表达改变的异构体 TGF¿然后，将使用受调节的基因来评估基因表达是否随着每个特定受体被破坏而改变。这些数据之间的相关性将指向介导 EMT 的特定受体和配体信号复合物。其他实验室的数据表明心脏 EMT 中存在多种信号转导机制和细胞内调节因子，包括 ErbB2、NF-1、VEGF 和 NFATc1。在第二个目标中，将进行实验来确定。是否破坏 TGF¿信号传导会改变任何其他候选机制，或者这些分子的丢失是否会改变 TGF¿在神经嵴系统中观察到，Sox 8、9或10在神经管中的强制表达会引起EMT并产生异位神经嵴细胞。心脏，它需要一些 TGF¿第三个目标是通过实验来确定 Sox 基因的外源表达是否会强制发生 EMT，以及 Sox 基因的表达是否受到 TGF 的调节。信号转导或调节 TGF¿这些实验将共同揭示 EMT 调节机制并有助于了解先天性心脏病。