GENE TRANSFER OF FIBRINOLYTIC AGENTS IN LUNG FIBROSIS

肺纤维化中纤溶剂的基因转移

• 批准号: 6110715
• 负责人: Richard H Simon
• 金额: $ 25.55万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-12-01 至 1999-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6110715
• 关键词: blood coagulation; fibrinolysis; gene therapy; human tissue; inflammation; laboratory mouse; nonhuman therapy evaluation; plasminogen activator inhibitors; pulmonary fibrosis /granuloma; tissue /cell culture; urokinase

The spectrum of pathogenic processes that leads to pulmonary fibrosis represents a number of parallel, redundant pathways. A therapeutic intervention that targets a single step common to multiple pathways would be a welcome addition to the currently dismal state of treatment options for fibrotic lung diseases. During many inflammatory lung diseases, fibrin accumulates within the alveolar compartment, due in part to impairment of the normally fibrinolytic activity of the alveolar space. The persistence of fibrin has pathologic importance because it can serve as a scaffold on which fibroblasts invade to form collagenous scars. Using transgenic mice, we have recently found that the amount of fibrosis induced by intratracheal bleomycin is strongly influenced by the activity of the fibrinolytic system. Transgenic mice with suppressed fibrinolytic activity from over-expression of a murine plasminogen activator inhibitor-1 (PAI-1) transgene develop increased fibrosis, while mice with inactivated PAI-1 genes are relatively protected. These results, combined with the observations of others, have encouraged us to explore the strategy of increasing plasminogen activation in the alveolar space as a means to limit fibrosis during pulmonary inflammation. Although a number of approaches could be employed, we have elected to use gene transfer technology as the means to augment fibrolysis. Our experience with this modality leads us to conclude that pulmonary directed gene transfer using recombinant adenovirus-based vectors can supply the means to test our Hypothesis: Enhancement of fibrinolytic activity within the alveolar space using gene transfer technology will reduce the pulmonary fibrosis that accompanies inflammatory lung injury. We propose three Specific Aims: 1) Transfer genes for urokinase-type plasminogen activator (uPA) and PAI- 1-resistant uPA to human and murine cells in vitro and determine the effects on cell-mediated plasminogen activation and fibrin matrix degradation. 2) Transfer genes for uPA and PAI-1-resistant uPA to the lungs of mice and determine the effects on plasminogen activator activity and fibrin degradation within the alveolar space. 3) Transfer genes for uPA and PAI-1-resistant uPA to the lungs of mice and determine the effects on pulmonary fibrosis induced by inflammation. In addition to evaluating a novel therapeutic strategy for fibrotic lung diseases, our studies will provide valuable information on the relationship between fibrinolysis and fibrogenesis, and on in vivo gene transfer to the distal airspaces. A further benefit will be the provision of gene transfer technology to other Projects of this SCOR (see Project 5).

导致肺纤维化的致病过程谱 代表了许多平行的、冗余的路径。治疗 针对多个途径共同的单个步骤的干预将 是对目前令人沮丧的治疗选择状态的一个受欢迎的补充 治疗纤维化肺病在许多炎症性肺病中， 纤维蛋白积聚在肺泡室中，部分原因是 肺泡腔的正常纤溶活性受损。 纤维蛋白的持续存在具有病理重要性，因为它可以 作为成纤维细胞侵入形成胶原性瘢痕的支架。 使用转基因小鼠，我们最近发现， 博莱霉素诱导的细胞毒活性受 纤维蛋白溶解系统。纤溶抑制转基因小鼠 来自鼠纤溶酶原激活剂的过表达的活性 抑制剂-1（派-1）转基因小鼠发生纤维化增加， 失活的派-1基因相对受到保护。这些结果，结合 与其他人的观察，鼓励我们探索 增加肺泡腔中纤溶酶原活化的策略， 一种在肺部炎症期间限制纤维化的方法。虽然 可以采用多种方法，我们选择使用基因 转移技术作为增加纤维溶解的手段。我们的经验 这种方式使我们得出结论，肺定向基因， 使用重组腺病毒载体的转移可以提供 为了验证我们的假设：增强纤维蛋白溶解活性， 利用基因转移技术的肺泡间隙将减少肺 伴随炎性肺损伤的纤维化。 我们提出三个具体目标： 1)尿激酶型纤溶酶原激活物（uPA）和PAI-1的转移基因 1-抗uPA的人和小鼠细胞体外，并确定 对细胞介导的纤溶酶原激活和纤维蛋白基质的影响 降解 2)将uPA和派-1抗性uPA的基因转移到小鼠的肺中 并测定对纤溶酶原激活物活性和纤维蛋白的影响 在肺泡腔内降解。 3)将uPA和派-1抗性uPA的基因转移到小鼠的肺中 并确定对炎症诱导的肺纤维化的影响。 除了评估纤维化肺的新治疗策略外， 疾病，我们的研究将提供宝贵的信息， 纤溶与纤维化的关系及体内基因研究 转移到远端的空气空间。另一个好处是， 基因转移技术的其他项目（见项目 5）。