Mechanisms for Has2 activation and hyaluronan-mediated cellular invasion

Has2 激活和透明质酸介导的细胞侵袭机制

• 批准号: 7799760
• 负责人: Evisabel A Craig
• 金额: $ 2.23万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-04-15 至 2010-09-17
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7799760
• 关键词: Binding; Biological Assay; Cardiac; Cell Proliferation; Cells; Cessation of life; Cloning; Congenital Heart Defects; Data; Development; Diagnostic; Early Diagnosis; Embryo; Embryonic Development; Event; Extracellular Matrix; Family member; Fibroblasts; Goals; Growth; Heart; Heart Diseases; Heart Valves; Hyaluronan; Immunoprecipitation; Integral Membrane Protein; Knowledge; Lead; Mediating; Modification; Molecular; Morphogenesis; Mus; Process; Production; Protein Family; Research; Research Proposals; Role; Series; Signal Pathway; Signal Transduction; Signaling Protein; Small Interfering RNA; Snails; Structure; System; Techniques; Testing; Time; United States; Western Blotting; base; cardiogenesis; cell behavior; cell motility; effective therapy; epithelial to mesenchymal transition; hyaluronan synthase 1; innovation; malformation; research study; snail protein; tool

DESCRIPTION (provided by applicant): Every year, approximately 50 out of 1,000 babies are born with a congenital heart defect (CHD) in the United States. Our long range goal is to understand the signaling mechanisms involved in heart septation and valve formation as miscues during these events constitute the most common cause of CHD. The heart valves and septae arise from molecular interactions between cardiac cushion cells and extracellular matrix components such as hyaluronan (HA). Throughout development, HA is primarily synthesized by the transmembrane protein hyaluronan synthase-2 (Has2). Thus, the objective of this application is to determine the role of key signaling proteins in Has2 activation and HA mediated cell invasion. HA can activate signaling effectors such as ERK and NFkB which, at the same time, have been shown to induce important regulators of cell behavior such as the Snail proteins. However, to this point no functional connection has been established between HA, NFkB and Snail proteins and the promotion of cellular invasion, a crutial process during cardiac development. Hence, our central hypothesis is that the activation of Has2 and subsequent production of HA leads to cell invasion via the induction of NFkB and Snail family members. We propose the following two specific aims to test our hypothesis: 1. Define the molecular modifications and interactions that regulate Has2 activity. 2. Determine whether HA signals through NFkB and Snail to induce invasive cellular migration. For our studies, we will utilize mouse embryonic fibroblast cells to perform a variety of experiments such as NFkB activity assays, real-time PCR, Western blots, immunoprecipitations, cloning, siRNA techniques, invasion and proliferation assays, etc. Our research will define the convergence of important signaling pathways for heart development through the actions of Has2 and its product hyaluronan. Understanding how these molecules exert their actions is crucial to help uncover the underlying causes for congenital heart defects. With this knowledge we expect to facilitate the development of diagnostic tools and more effective treatment for these cardiac diseases.

描述（由申请人提供）：每年，在美国，大约有1000名婴儿中约有50名患有先天性心脏缺陷（CHD）。我们的远距离目标是了解这些事件期间的错误和瓣膜形成所涉及的信号传导机制构成了CHD的最常见原因。心脏瓣膜和隔膜来自心脏缓冲细胞和细胞外基质成分（例如透明质酸（HA））之间的分子相互作用。在整个开发过程中，HA主要由跨膜蛋白透明质酸合酶2（HAS2）合成。因此，该应用的目的是确定关键信号蛋白在HAS2激活和HA介导的细胞侵袭中的作用。 HA可以激活诸如ERK和NFKB之类的信号传导效应子，同时已被证明会诱导细胞行为的重要调节剂，例如蜗牛蛋白。但是，到目前为止，HA，NFKB和蜗牛蛋白之间尚未建立功能连接，并且促进细胞侵袭，这是心脏发育过程中的残酷过程。因此，我们的核心假设是，HAS2的激活和随后的HA产生通过诱导NFKB和Snail家族成员导致细胞侵袭。我们提出以下两个特定目的来检验我们的假设：1。定义调节HAS2活性的分子修饰和相互作用。 2。确定HA是否通过NFKB和蜗牛发出信号以诱导侵入性细胞迁移。在我们的研究中，我们将利用小鼠胚胎成纤维细胞进行多种实验，例如NFKB活性测定，实时PCR，蛋白质印迹，免疫沉淀，克隆，克隆，siRNA技术，入侵和增殖试验等等。我们的研究将通过hys 2和ITS HYAR的速度来定义对心脏开发的重要信号途径的研究。了解这些分子如何采取行动对于帮助揭示先天性心脏缺陷的根本原因至关重要。有了这些知识，我们期望促进诊断工具的开发和对这些心脏病的更有效治疗。