Bone Marrow Derived Stromal Cells in Rat Models of Lung Injury

肺损伤大鼠模型中的骨髓衍生基质细胞

• 批准号: 7682214
• 负责人: Arnold Ralph Brody
• 金额: $ 29.29万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7682214
• 关键词: Adult; Alveolar; Alveolar Duct; Anatomy; Asbestos; Asbestos-related lung injury; Bone Marrow; Bone Marrow Cells; Bone Marrow Transplantation; Breathing; Cell physiology; Cells; Characteristics; Data; Data Analyses; Development; Disease; Endothelial Cells; Epithelial; Epithelial Cells; Epithelium; Event; Exposure to; Female; Fiber; Green Fluorescent Proteins; Individual; Injury; Lesion; Lung; Lung diseases; Marrow; Mesenchymal; Modeling; Nude Mice; Numbers; Phenotype; Play; Population; Process; Rattus; Research; Research Personnel; Role; Staging; Stem cells; Stromal Cells; Structure of parenchyma of lung; Surface; System; Testing; Therapeutic; Tissues; Trachea; Transgenic Organisms; Work; Y Chromosome; airway remodeling; alveolar epithelium; design; injured; interest; lung injury; male; repaired

This research is designed to test the central hypothesis that bone marrow-derived stromal stem cells engraft in the lung and differentiate to alveolar and airway epithelial and mesenchymal cells, consequently ameliorating or exacerbating the progression of fibroproliferative lung disease. Two well-characterized models of lung injury will be used: 1) Brief exposure to inhaled asbestos fibers rapidly causes injury of the bronchiolar-alveolar epithelium and a fibroproliferative lesion at the alveolar duct bifurcations. Studies were carried out in female rats that have undergone whole bone marrow transplantation from male green fluorescent protein (GFP)-transgenic syngeneic rats. The Preliminary Data shown here demonstrate that there are significantly increased numbers of GFP-positive, Y chromosome-positive bone marrow-derived cells in the asbestotic lesions compared with surrounding, uninjured tissues and with lungs from unexposed rats. Furthermore, the GFP-positive cells in the lung had both epithelial and mesenchymal phenotypes. Studies were also carried out in female rats injected intravascularly with 5 x 10[6] marrow stromal stem cells isolated from male GFP-transgenic rats. Our Preliminary Data demonstrate putative stem cells apparently attached to alveolar duct walls. Here we propose to define the subpopulations of stem cells that engraft and differentiate in injured lung and the effect they have on progression of asbestos-induced fibroproliferative lesions. 2) Denuded rat tracheae grafted into nude mice provide a milieu in which an epithelial lining can be reestablished, the characteristics of which is primarily determined by the differentiation potential of the inoculated cells. Our Preliminary Data show that purified populations of airway epithelial cells mixed with bone marrow-derived cells form a differentiated epithelium in the tracheal graft system. Furthermore the bone marrow-derived cells in the graft appear to differentiate into epithelial cells, as well as endothelial cells that participate in revascularization of the grafts. Some stem cells appear to differentiate as mesenchymal cells. This system will be ideal for testing the postulate since varying numbers of specific stem cell subpopulations in different stages of differentiation can be studied in combination with varying numbers and types of epithelial cells. There are no data available which allow us to predict whether or not the stem cells will play any role in the degree of lesion development or participate in populating the tracheo-bronchial and bronchiolar-alveolar duct walls. These studies will provide opportunities to develop therapeutic approaches through the use of adult bone marrow-derived stem cells because we know the precise anatomic and temporal distribution of the developing lesions. The stem cells will be characterized in, and we will be using the same populations of cells employed by investigators in the other projects. This will be a clear advantage for the final data analysis.

本研究旨在验证一个中心假设，即骨髓来源的基质干细胞植入肺内，并分化为肺泡和呼吸道上皮细胞和 间质细胞，从而改善或加剧纤维增生性肺病的进展。将使用两种典型的肺损伤模型：1)短暂暴露于吸入石棉纤维会迅速导致细支气管肺泡上皮损伤和肺泡管分叉处纤维增生性病变。研究是在从绿色荧光蛋白(GFP)转基因同基因大鼠进行全骨髓移植的雌性大鼠身上进行的。初步数据显示，与石棉样皮损相比，石棉样皮损中绿色荧光蛋白阳性、Y染色体阳性的骨髓来源细胞数量显著增加。 周围未受损伤的组织，以及未暴露的大鼠的肺。此外，肺中的GFP阳性细胞既有上皮型也有间叶型。从雄性GFP转基因大鼠分离的5x10[6]骨髓基质干细胞血管内注射的雌性大鼠也进行了研究。我们的初步数据显示，假定的干细胞明显附着在肺泡管壁上。在这里，我们建议确定在受损肺中植入和分化的干细胞亚群，以及它们在石棉诱导的纤维增生性病变进展中的作用。2)裸大鼠气管移植到裸鼠体内提供了重建上皮衬里的环境，其特点是 主要由接种细胞的分化潜力决定。我们的初步数据显示，在气管移植系统中，纯化的呼吸道上皮细胞群与骨髓来源的细胞混合形成分化的上皮细胞。此外，移植物中的骨髓来源细胞似乎分化为上皮细胞，以及参与移植物血管重建的内皮细胞。一些干细胞似乎分化为间充质细胞。这个系统将是测试假设的理想选择，因为不同分化阶段不同数量的特定干细胞亚群可以 结合不同数量和类型的上皮细胞进行研究。没有可用的数据可以让我们预测干细胞是否会在 病变发展程度或参与填充气管-支气管壁和细支气管-肺泡壁。这些研究将提供通过使用成人骨髓来源的干细胞来开发治疗方法的机会，因为我们知道正在发展的病变的精确解剖和时间分布。干细胞将在中进行表征，我们将使用其他项目中研究人员使用的相同群体的细胞。这将是最终数据分析的明显优势。