RESPONSE IN LUNG EXTRACELLULAR MATRIX TO ASBESTOS

肺细胞外基质对石棉的反应

• 批准号: 7959561
• 负责人: Elizabeth A Putnam
• 金额: $ 10.78万
• 依托单位: UNIVERSITY OF MONTANA
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7959561
• 关键词: Address; Amphiboles; Asbestos; Asbestosis; Attention; Breathing; Cell Communication; Computer Retrieval of Information on Scientific Projects Database; Crocidolite Asbestos; Deposition; Development; Disease; Elements; Environmental Health; Exposure to; Extracellular Matrix; Extracellular Matrix Proteins; Fiber; Fibroblasts; Fibrosis; Funding; Gene Expression; Goals; Grant; Growth Factor; Health; Individual; Institution; Investigation; Knockout Mice; Lung; Lung diseases; Malignant neoplasm of lung; Measures; Mesothelioma; Mus; Process; Production; Proteins; Public Health; RNA Interference; Regulation; Research; Research Personnel; Resources; Risk; Role; Saline; Site; Source; Testing; Therapeutic; United States National Institutes of Health; Wild Type Mouse; extracellular; in vivo; lung development; new therapeutic target; response; vermiculite

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Attention to asbestos-related diseases (ARD) has intensified due to the exposure of Libby, MT residents to asbestos-contaminated vermiculite. Vermiculite distribution to 200+ sites nationwide and the long latent period for disease development make ARD a continuing public health issue. Exposures to asbestos fibers by inhalation cause multiple ARD including asbestosis, lung cancer and mesothelioma. This investigation will focus on asbestos-caused fibrosis by studying the role of SPARC in this process, since we found increased expression of SPARC in mouse lungs exposed to several types of asbestos fibers. SPARC is a matricellular protein involved in the regulation of extracellular matrix (ECM)cell interactions through modulation of growth factor activity. These previously identified functions make SPARC an exciting candidate for involvement in asbestosis. The investigations proposed here will elucidate the role of SPARC in asbestosis, specifically targeting how inhibition of SPARC expression can regulate ECM production. The goals of this project are to delineate the involvement of the matricellular protein SPARC in the regulation of ECM deposition in response to crocidolite and the Libby amphibole. The central hypothesis to be tested in our studies is that expression of SPARC is a significant step in the development of lung fibrosis through the modulation of ECM production. To test the hypothesis, we propose the following aims: 1) Establish the in vivo expression of SPARC and ECM proteins in Sparc-null and matched wild-type (WT) mice after exposure to saline, the Libby amphibole, and crocidolite asbestos. Key elements of this aim include the use of Sparc-null mice constitutively lacking the protein as well as RNA interference (RNAi) as a mechanism to control SPARC expression in WT mice after fiber exposure. 2) Characterize the response of primary lung fibroblast cultures isolated from Sparc-null and matched WT mice to saline, the Libby amphibole, and crocidolite asbestos. Key elements of this aim include analysis of ECM proteins as well as changes in gene expression after amphibole exposure. As a result of the proposed studies, we hope to achieve a better understanding of the role of SPARC in the development of lung fibrosis as measured by ECM production after amphibole exposure. We will also study the potential for blocking SPARC expression as a therapeutic measure to reduce the fibrotic response. By investigating a critical question concerning fibrosis development, this proposal will also begin to address the potential for using RNA interference to control SPARC expression as a treatment for fibrotic diseases of the lung. Because of the health risks faced by thousands of asbestos-exposed individuals, the ultimate goal of these studies is to identify a direction for the development of novel therapeutic targets for these and other similar environmentally-caused lung diseases. In addition, this potential therapy has implications for treatment of all forms of fibrosis, including IPF.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 由于蒙大拿州利比市居民接触受石棉污染的蛭石，人们对石棉相关疾病 (ARD) 的关注不断加强。 蛭石分布于全国 200 多个地点，疾病发展潜伏期长，使急性呼吸道疾病成为持续的公共卫生问题。 吸入石棉纤维会导致多种急性呼吸道疾病，包括石棉肺、肺癌和间皮瘤。这项研究将通过研究 SPARC 在这一过程中的作用来重点关注石棉引起的纤维化，因为我们发现暴露于几种类型石棉纤维的小鼠肺部 SPARC 表达增加。 SPARC 是一种基质细胞蛋白，通过调节生长因子活性参与调节细胞外基质 (ECM) 细胞相互作用。这些先前确定的功能使 SPARC 成为参与石棉沉着症的令人兴奋的候选者。本文提出的研究将阐明 SPARC 在石棉沉着症中的作用，特别针对 SPARC 表达的抑制如何调节 ECM 的产生。该项目的目标是描述基质细胞蛋白 SPARC 在响应青石棉和利比角闪石的 ECM 沉积调节中的参与。我们研究中要测试的中心假设是 SPARC 的表达是通过调节 ECM 产生而导致肺纤维化发展的重要一步。 为了检验这一假设，我们提出以下目标：1) 在暴露于盐水、利比角闪石和青石棉后，在 Sparc-null 和匹配的野生型 (WT) 小鼠中建立 SPARC 和 ECM 蛋白的体内表达。这一目标的关键要素包括使用本质上缺乏该蛋白质的 Sparc-null 小鼠以及 RNA 干扰 (RNAi) 作为控制纤维暴露后 WT 小鼠 SPARC 表达的机制。 2) 表征从 Sparc-null 和匹配的 WT 小鼠分离的原代肺成纤维细胞培养物对盐水、Libby 角闪石和青石棉的反应。该目标的关键要素包括 ECM 蛋白分析以及角闪石暴露后基因表达的变化。 作为拟议研究的结果，我们希望更好地了解 SPARC 在肺纤维化发展中的作用（通过角闪石暴露后 ECM 的产生来测量）。我们还将研究阻断 SPARC 表达作为减少纤维化反应的治疗措施的潜力。通过研究有关纤维化发展的关键问题，该提案还将开始探讨使用 RNA 干扰控制 SPARC 表达作为肺部纤维化疾病治疗的潜力。由于成千上万接触石棉的人面临着健康风险，这些研究的最终目标是为这些和其他类似的环境引起的肺部疾病确定新的治疗靶点的开发方向。此外，这种潜在的疗法对治疗所有形式的纤维化（包括特发性肺纤维化）具有重要意义。