DYNAMIC INTERACTION OF ECM AND CARDIAC FIBROBLAST

描述：（改编自研究者摘要） 心脏的功能是由大的肌细胞决定的， 心肌的大部分体积和细胞外基质 （ECM）。 心脏成纤维细胞是参与心肌纤维化的主要细胞类型。 ECM的动态调节，也是ECM中的主要细胞类型。 心肌占细胞的大约90%。 这项建议 解决了成纤维细胞功能受以下因素调节的总体假设： 特异性细胞表面受体（整联蛋白）之间的动态相互作用， 成纤维细胞的内在遗传调节和细胞的结构， ECM本身。 成纤维细胞经历表型变化， 在发育过程中的生理信号，导致的变化， 企业内容管理的组成和组织。 然而， 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， 不同的生理阶段（年龄），以检查整体假设， ECM和成纤维细胞之间的动态相互作用对于 成纤维细胞的调节，从而组织， ECM的组成。 具体而言，假设合成 心脏成纤维细胞的胶原蛋白的表达与 MMP和胶原特异性整合素。 本提案的具体目标 是：1）检测特异性整合素、金属蛋白酶 (MMP)在胎儿、新生儿和成人发育过程中， 2)在体外分离的心脏成纤维细胞中检查这些相同的参数 从相同的生理阶段; 3）改变整联蛋白的表达， MMPs和胶原合成在分离的成纤维细胞中的作用 寡核苷酸和腺病毒转导，以确定 这3个参数; 4）确定机械刺激对 心肌细胞整合素、胶原和基质金属蛋白酶的表达和积累 不同生理阶段的成纤维细胞;和5）修饰结构 体外分析ECM成分，以确定异常的ECM是否会影响 整合素、MMPs和胶原蛋白的表达。 这些研究将利用 多种形态学、生物化学和分子技术分析 ECM和基因表达之间的动态相互作用， 导致组成变化的关键生理阶段， ECM的组织。 然而，这些表型的机制 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成纤维细胞从不同的生理 阶段，以检查整体假设，即动态相互作用 细胞外基质和成纤维细胞之间的相互作用对于病毒的传播至关重要 机械和化学信号，这是必不可少的调节 开发过程中ECM的组织和组成。 具体来说就是 假设心脏成纤维细胞胶原蛋白的合成是 与MMP和胶原特异性整合素的表达协调。

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.