DYNAMIC INTERACTION OF ECM AND CARDIAC FIBROBLAST

描述：(改编自《调查员摘要》)表格和 心脏的功能由组成心脏的大肌细胞决定。 心肌和细胞外基质的大部分体积 (ECM)。心脏成纤维细胞是参与 细胞外基质的动态调节，也是细胞外基质的主要细胞类型 心肌约占细胞总数的90%。这项建议 提出成纤维细胞功能受以下因素调节的总体假设 特定细胞表面受体(整合素)之间的动态相互作用 成纤维细胞的内在遗传调控和细胞的结构 ECM本身。成纤维细胞经历表型变化以响应 发育过程中的生理信号，导致大脑 ECM的组成和组织。然而，其机制是如何 这些表型变化对成纤维细胞功能的影响尚不清楚。在 研究人员使用分离的心脏成纤维细胞进行拟议研究 不同的生理阶段(年龄)来检验总体假设 细胞外基质和成纤维细胞之间的动态相互作用对于 对成纤维细胞的调节，从而调节组织和 ECM的组成。具体地说，它假设合成 心脏成纤维细胞对胶原蛋白的表达与 MMPs和胶原特异性整合素。这项建议的具体目的 目的是：1)检测特定整合素、金属蛋白酶的表达 胎儿、新生儿和成人发育过程中体内的基质金属蛋白酶和胶原蛋白； 2)在体外分离的心脏成纤维细胞中检测这些相同的参数 3)改变整合素的表达， 反义MMPs与成纤维细胞胶原合成的关系 寡核苷酸和腺病毒转导相互作用的研究 这3个参数：4)决定了机械刺激对运动能力的影响。 心脏整合素、胶原蛋白和基质金属蛋白酶的表达和积聚 不同生理阶段的成纤维细胞；5)改变结构 以确定异常的细胞外基质是否会影响细胞外基质。 整合素、基质金属蛋白酶和胶原蛋白的表达。这些研究将利用 用于分析的各种形态、生化和分子技术 细胞外基质与基因表达的动态相互作用 关键的生理阶段，导致成分和 ECM的组织。然而，这些表型如何表现的机制 变化对成纤维细胞功能的影响尚不清楚。在拟议的研究中， 研究人员将使用分离的不同生理来源的成纤维细胞 检验动态相互作用的总体假设的阶段 在细胞外基质和成纤维细胞之间是至关重要的传递 机械和化学信号对调节 ECM在开发过程中的组织和组成。具体地说，它是 假设心脏成纤维细胞合成胶原是 与MMPs和胶原特异性整合素的表达相协调。

DESCRIPTION: (Adapted from the Investigator's Abstract) The form and function of the heart is determined by the large myocytes which comprise the majority of the volume of the myocardium and by the extracellular matrix (ECM). The cardiac fibroblast is the primary cell type involved in the dynamic regulation of the ECM and is also the dominant cell type in the myocardium accounting for approximately 90% of the cells. This proposal addresses the overall hypothesis that fibroblast function is regulated by the dynamic interaction between specific cell surface receptors (integrins), the intrinsic genetic regulation of the fibroblast and the structure of the ECM itself. The fibroblast undergoes phenotypic changes in response to physiological signals during development that result in changes in the composition and organization of the ECM. However, the mechanisms of how these phenotypic changes affect fibroblast function is not clear. In the proposed studies the investigators use isolated cardiac fibroblasts from different physiological stages (ages) to examine the overall hypothesis that the dynamic interaction between the ECM and the fibroblast is essential for the regulation of the fibroblast and consequently the organization and composition of the ECM. Specifically, it is hypothesized that the synthesis of collagen by cardiac fibroblasts is coordinated with the expression of MMPs and collagen specific integrins. The specific aims of this proposal are to: 1) examine the expression of specific integrins, metalloproteases (MMP), and collagen in vivo during fetal, neonatal, and adult development; 2) examine these same parameters in vitro in cardiac fibroblasts isolated from the same physiological stages; 3) alter the expression of integrins, MMPs, and collagen synthesis in isolated fibroblasts by using antisense oligonucleotides and adenoviral transduction to determine the interaction of these 3 parameters; 4) determine the effect of mechanical stimulation on the expression and accumulation of integrins, collagens and MMPs of cardiac fibroblasts at different physiological stages; and 5) modify the structure of the ECM components in vitro to determine if abnormal ECM can affect the expression of integrins, MMPs and collagens. These studies will utilize a variety of morphological, biochemical and molecular techniques to analyze the dynamic interaction between the ECM and gene expression at several critical physiological stages that result in changes in the composition and organization of the ECM. However, the mechanisms of how these phenotypic changes affect fibroblast function is not clear. In the proposed studies, the investigators will use isolated fibroblasts from different physiological stages to examine the overall hypothesis that the dynamic interaction between the ECM and the fibroblast is critical to the transmission of mechanical and chemical signals which are essential for the regulation of ECM organization and composition during development. Specifically, it is hypothesized that the synthesis of collagen by cardiac fibroblasts is coordinated with the expression of MMPs and collagen specific integrins.