LUNG CONNECTIVE TISSUE--RESPONSES TO INJURY AND REPAIR

肺间质弹性纤维和胶原纤维在肺间质纤维化中起关键作用 肺结构和功能。这些结缔组织中的异常 蛋白质被认为与阻塞性和纤维性肺部疾病有关。 这两种疾病的动物模型已被广泛应用于 在过去的几十年里定义了降解性物质的具体机制 和纤维性病变。尽管这些研究提供了有用的 关于病原体和组织反应的信息，细胞 而这些反应背后的分子机制在很大程度上是未知的。 该计划的总体目标是确定调控机制 肺间质结缔组织蛋白对条件的反应 模拟受伤/修复情况。我们建议研究这些机制 使用肺成纤维细胞的体外模型，我们可以减少 细胞内外调节反应的研究 整个肺组织的复杂性与细胞的相对简单性 文化体系。细胞培养的使用将使我们能够控制 损伤/扰动暴露的程度和修复/反应的参数。 涉及的人员包括高级和初级教职员工以及 主要员工的概念和培训经验和培训 生物化学、呼吸生理学和 分子生物学、细胞生物学和发育生物学在我们的 拟开展的研究。研究的基础是建立在强大的核心之上的 他们的专业知识和资源结合在一起创造了 一支独特而强大的团队，负责解剖肺内潜在的突起 结缔组织损伤与修复。 我们研究这些机制的计划分为五个，高度 综合项目。项目1侧重于调节弹性蛋白和 微原纤维基因在基质蛋白降解条件下的表达。 研究旨在探索降解产物、基质的作用 以及可能的内生生长因素控制上下- 原弹性蛋白基因表达水平的调控。项目2侧重于机制 由此，弹性蛋白和胶原基因的表达被效应器下调 物质。总体目标是确定独联体的行动和 调节这一调节的反式作用因子。项目3的重点是 伴随着基质蛋白分解的细胞外弹性反应 使用了各种蛋白酶和曝光条件。研究的目的是 确定弹性蛋白重新合成(修复)的机制。项目4重点 多肽因子对赖氨酰氧化酶基因表达的调节作用 改变其弹性蛋白和胶原蛋白表达的环境扰动 底物。项目5的重点是不同的多肽单位的作用 原弹性蛋白在引导和排列分子形成不溶物质中起作用 弹性蛋白和调节弹性蛋白基因的表达。这些项目 由提供基本细胞的三个核心单元支持 自动培养、分子、生化和免疫探针 分析、显微镜和行政支持。

Elastic and collagenous fibers of lung interstitium play key roles in pulmonary structure and function. Abnormalities in these connective tissue proteins have been implicated in obstructive and fibrotic lung diseases. Animal models of both types of disease have been used extensively in the last several decades to define specific mechanisms underlying degradative and fibrotic pathologies. Although these studies have provided useful information regarding causative agents and tissue responses, the cellular and molecular mechanisms underlying these responses are largely unknown. The overall goal of this program is to determine the mechanisms regulating pulmonary interstitium connective tissue protein responses to conditions mimicking injury/repair situations. We propose to study these mechanisms using an in vitro model of pulmonary fibroblast cells where we can reduce studies of intra- and extracellular regulatory responses from the complexity of whole lung tissue to the relative simplicity of a cell culture system. The use of cell cultures will allow us to control the extent of injury/perturbant exposure and the parameters of repair/response. The personnel involved include both senior and junior faculty as well as key staff members whose experience and training in the conceptual and technological aspects of biochemistry, respiratory physiology and molecular, cellular and developmental biology are integrated throughout our proposed research. The research foundation is built upon a strong nucleus of established investigators whose combined expertise and resources create an unique and powerful team for dissecting processes underlying pulmonary connective tissue injury and repair. Our plan for investigating these mechanisms is divided among five, highly integrated projects. Project 1 focuses on the regulation of elastin and microfibril gene expression under conditions of matrix proteolysis. Studies are designed to explore the role of degradation products, matrix and possible endogenous growth factors in controlling up and down- regulation of tropoelastin mRNA levels. Project 2 focuses on mechanisms whereby elastin and collagen gene expression is down regulated by effector substances. The overall objective is to determine the actions of cis and trans acting factors in modulating this regulation. Project 3 focuses on the extracellular elastogenic responses that accompany matrix proteolysis using a variety of proteases and exposure conditions. Studies are aimed at determining mechanisms of elastin resynthesis (repair). Project 4 focuses on the regulation of lysyl oxidase gene expression by peptide factors and environmental perturbants that alter expression of its elastin and collagen substrates. Project 5 focuses on the role that various peptide units of tropoelastin play in directing and aligning the molecule to form insoluble elastin and in the regulation of elastin gene expression. These projects are supported by three core units which provide the essential cell cultures, molecular, biochemical and immunological probes, automated analyses, microscopy and administrative support.