EFFECTS OF ESTROGEN ON OSTEOCLASTS FROM TRANSGENIC MICE

雌激素对转基因小鼠破骨细胞的影响

• 批准号: 3161728
• 负责人: Garson DAVID ROODMAN
• 金额: $ 17.56万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCIENCE CENTER
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-30 至 1995-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3161728
• 关键词: biotechnology; calcitonin; cell line; clone cells; estrogens; gene expression; genetic library; genetic manipulation; genetic promoter element; genetically modified animals; laboratory mouse; microinjections; osteoclasts; osteoporosis; pathologic bone resorption; phenotype; regulatory gene; tissue /cell culture

Little information is available on factors directly controlling the osteoclast or the molecular events responsible for osteoclast formation and bone resorption. The cell and molecular biological characterization of the osteoclast has been severely hampered by the inability to isolate large numbers of osteoclasts or osteoclast precursors and the unavailability of established osteoclastic cell lines. The proposed studies are designed to allow the establishment of cell lines that express the full differentiated phenotype of osteoclasts from osteoclast tumors formed in transgenic mice. These cell lines will then be used to address questions concerning the effects of estrogen and other osteotropic factors on osteoclast function. The proposed strategy involves directing the expression of an oncogene, such as SV40 Tantigen, specifically or preferentially to osteoclast cells using the regulatory regions of the genes encoding tartrate resistant acid phosphatase (TRAP) and the calcitonin receptor (CTR). Transgenic mice expressing these hybrid transgenes are expected to develop bone tumors of osteoclast origin, or possibly immortalized osteoclast precursors in the blood, spleen or bone marrow. These immortalized cells will be placed in culture and cell lines will be established. The cell lines will be extensively characterized to determine the extent to which they resemble osteoclasts or osteoclast precursors and to test their ability to differentiate in culture. In addition, cell lines will be derived from both male and female mice to allow a comparison of phenotype. Finally, since estrogen may affect the formation and activity of osteoclasts, studies will be undertaken to determine the effects of estrogen on the growth and differentiation of these cells and on their ability to resorb bone. In addition, estrogen will be administered to the transgenic mice to determine if the pattern of mononuclear or multinucleated osteoclastic cells is altered in vivo. Based on the success of the transgenic mouse approach for deriving established differentiated cell lines representing other rare cell types, we are hopeful that the proposed studies will lead to the development of established cell lines representing osteoclasts or osteoclast precursors. Such cell lines would provide a novel system for in vitro analysis of important questions concerning the role of estrogen and other osteotropic factors or accessory cells in regulating osteoclast function.

关于直接控制气候变化的因素的资料很少。 破骨细胞或负责破骨细胞形成的分子事件， 骨吸收 本研究对该菌的细胞和分子生物学特性进行了研究。 破骨细胞由于不能分离大的 破骨细胞或破骨细胞前体的数量以及 建立成骨细胞系。 拟议的研究旨在 允许建立表达完全分化的 转基因小鼠中形成的破骨细胞肿瘤的破骨细胞表型。 然后，这些细胞系将被用于解决关于 雌激素和其他促骨因子对破骨细胞功能的影响。 所提出的策略包括指导癌基因的表达， 如SV40 T抗原，特异性或优先针对破骨细胞 利用编码抗酒石酸的基因的调节区 磷酸酶（TRAP）和降钙素受体（CTR）。 转基因小鼠 表达这些杂合转基因预计会产生骨肿瘤， 破骨细胞起源，或可能永生化的破骨细胞前体， 血液脾脏或骨髓这些永生化细胞将被放置在 将建立培养物和细胞系。 细胞系将是 以确定它们在多大程度上 破骨细胞或破骨细胞前体，并测试其 文化上的差异。 此外，细胞系将来源于 雄性和雌性小鼠以比较表型。 最后，由于雌激素可能影响 破骨细胞，将进行研究，以确定 雌激素对这些细胞的生长和分化以及它们的 吸收骨的能力。 此外，雌激素将被施用到 转基因小鼠，以确定是否单核细胞或 多核骨细胞在体内发生改变。 基于转基因小鼠方法的成功， 建立了代表其他稀有细胞类型的分化细胞系， 我们希望拟议的研究将导致发展 建立代表破骨细胞或破骨细胞前体的细胞系。 这样的细胞系将提供一种用于体外分析的新系统， 关于雌激素和其他骨促进剂作用的重要问题 因子或辅助细胞调节破骨细胞功能。