Mineralization: Integrins and Focal Adhesion Kinase

矿化：整合素和粘着斑激酶

• 批准号: 6478029
• 负责人: GALEN B SCHNEIDER
• 金额: $ 7.35万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至 2004-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6478029
• 关键词: atomic absorption spectrometry; calcium; cell adhesion; cell differentiation; cell growth regulation; collagen; cytoskeleton; developmental genetics; extracellular matrix; focal adhesion kinase; gene expression; genetic regulation; immunologic techniques; integrins; intermolecular interaction; microscopy; osteoblasts; osteogenesis; polymerase chain reaction; protein structure function; receptor coupling; statistics /biometry; tissue /cell culture; transcription factor; transfection; western blottings

Osteogenesis involves the recruitment of multi-potential mesenchymal stem cells and the progressive differentiation of these cells into osteoblasts. Osteoblast differentiation and skeletal formation during embryonic development is mediated by an essential transcription factor protein core binding core binding factor-alpha1 (cbfa1). Integrin receptors are widely expressed and bind extracellular matrix proteins. They also regulate intracellular signaling pathways that contribute to the development of cranial-facial structures, wound healing, cell motility, cell growth and differentiation, and apoptosis. Signaling pathways and proteins mediated by integrins, such as the focal adhesion tyrosine kinase FAK, can regulate cell growth and differentiation, as well as cell fate. Yet the role of integrin receptor mediation of osteoblast differentiation and subsequent mineralization is not fully understood. Adhesion- mediated signaling transduced through osteoblast-extracellular matrix interactions during osteogenesis may represent essential pathways for processing environmental cues necessary for gene expression, as well as bone formation and remodeling during osteoblast differentiation. The goal of this proposal is to determine if the integrin associated protein tyrosine kinase FAK mediates osteogenic gene expression (Cbfa1, BSP, Collagen type I), and subsequent mineralization. We hypothesize that blocking the interaction of various osteoblast integrin receptors with their underlying extracellular matrix with specific antibodies will affect gene expression and minerization. We also predict that over-expressing the integrin associated protein tyrosine kinase FAK in a non- mineralizing. We also predict that over-expressing the integrin associated protein tyrosine kinase FAK in a non-mineralizing OB clones will lead to mineralization. We will use histological, biochemical, and molecular approaches to perform these studies. Data acquired from these studies will allow us to better understand how osteoblast differentiation and osteogenesis are mediated through integrin receptors and associated signaling proteins such as FAK. This will allow for development of new therapeutic approaches that in turn could lead to novel treatments and tissue-engineered biological applications to increase the quality of life for many patients.

成骨涉及多潜能间充质干细胞的招募和这些细胞向成骨细胞的渐进分化。胚胎发育过程中的成骨细胞分化和骨形成是由必需转录因子蛋白核心结合核心结合因子α1(Cbfa1)介导的。整合素受体广泛表达并与细胞外基质蛋白结合。它们还调节细胞内的信号通路，这些信号通路有助于颅面结构的发育、伤口愈合、细胞运动、细胞生长和分化以及细胞凋亡。整合素介导的信号通路和蛋白，如粘着斑酪氨酸激酶FAK，可以调节细胞的生长和分化，以及细胞的命运。然而，整合素受体在成骨细胞分化和随后的矿化中的作用尚不完全清楚。在成骨过程中，通过成骨细胞-细胞外基质相互作用传递的黏附介导的信号可能是处理基因表达所需的环境信号以及成骨细胞分化过程中骨形成和重塑的重要途径。这项建议的目的是确定整合素相关蛋白酪氨酸激酶FAK是否介导成骨基因的表达(Cbfa1、BSP、I型胶原)以及随后的矿化。我们假设，用特定的抗体阻断各种成骨细胞整合素受体与其潜在的细胞外基质的相互作用将影响基因的表达和矿化。我们还预测整合素相关蛋白酪氨酸激酶FAK在非矿化中过表达。我们还预测，在非矿化OB克隆中过表达整合素相关蛋白酪氨酸激酶FAK将导致矿化。我们将使用组织学、生化和分子方法来进行这些研究。从这些研究中获得的数据将使我们更好地了解成骨细胞分化和成骨是如何通过整合素受体和相关信号蛋白如FAK介导的。这将允许开发新的治疗方法，进而可能导致新的治疗方法和组织工程生物应用，以提高许多患者的生活质量。