HORMONAL CONTROL OF CYTOKINES IN BONE AND MARROW CELLS

骨细胞和骨髓细胞中细胞因子的激素控制

• 批准号: 6098701
• 负责人: STAVROS C. MANOLAGAS
• 金额: $ 19.64万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-06-01 至 2000-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6098701
• 关键词: androgens; bone development; bone marrow; estrogens; genetic regulatory element; hormone regulation /control mechanism; immunocytochemistry; interleukin 6; laboratory mouse; monoclonal antibody; osteoblasts; osteoclasts; polymerase chain reaction; tissue /cell culture; transcription factor

Work leading to this application indicates that the IL-6 gene is transcriptionally regulated by both classes of sex steroids and plays a critical role in the bone loss of gonadal function; that estrogens and androgens also downregulate the expression of the ligand-binding and the signal transducing components of the IL-6 receptor; and that their loss enhances the responsiveness of osteoclast precursors to IL-6 and IL-11, and upregulates osteoblast development. Based on this evidence, it is proposed that IL-6, and other members of the cytokine subfamily that share gp130 for the transduction of their signals, are capable of stimulating not only osteoclast, but also osteoblast, development. The production of IL-6, as well as the action of IL-6 and the action of other members of this cytokine subfamily, is under the inhibitory control of sex steroids. Removal of these inhibitory effects contributes to the increased bone remodeling and the bone loss associated with loss of gonadal function. To test this hypothesis, the mechanism(s) underlying the downregulating effects of estrogens and androgens on the transcription of the genes for IL-6, the ligand-binding subunit of the IL-6 receptor (gp80), and the signal transducing subunit (gp130) will be elucidated by identifying the cis-acting regulatory elements involved and their corresponding transcription factors. Further, the effects of sex steroids on the production of the soluble IL-6 receptor will be studied and cells expressing the mRNAs for IL-6, gp80 and gp130 in ex vivo murine bone marrow cultures and undecalcified cancellous bone sections from sex steroid replete and sex steroid deficient mice will be identified, using in situ RT-PCR in combination with immunohistochemistry. In addition, the role of the IL-6 subfamily of cytokines in the upregulation of osteoblast progenitor formation in the bone marrow following loss of sex steroids, and potential interactions between these cytokines and other growth factors on osteoblastogenesis will be investigated. Finally, chimeric constructs of the soluble IL-6 or CNTF receptor alpha, complexed with monoclonal antibodies recognizing cell surface markers, will be used to selectively direct the corresponding cytokines to either osteoclast or osteoblast progenitors; and investigate the effects of such manipulation on osteoclastogenesis and osteoblastogenesis.

导致这一应用的工作表明，IL-6基因是 在转录水平上受两类性激素的调节，并发挥着 在性腺功能骨质丧失中的关键作用；雌激素和 雄激素还下调配体结合和 IL-6受体的信号传导成分；以及它们的丢失 增强破骨细胞前体对IL-6和IL-11的反应性， 并促进成骨细胞的发育。基于这个证据，它是 建议IL-6和其他共享细胞因子亚家族的成员 Gp130用于其信号的转导，都能够刺激 不仅是破骨细胞，成骨细胞也在发育。产品的生产 IL-6的作用以及IL-6的作用和其他成员的作用 这个细胞因子亚家族，受性类固醇的抑制控制。 这些抑制作用的消除有助于增加骨骼 与性腺功能丧失相关的重塑和骨质丢失。至 验证这一假设，下调调控背后的机制(S) 雌激素和雄激素对血管内皮生长因子基因转录的影响 IL-6，IL-6受体的配体结合亚单位(Gp80)，以及 信号转导亚单位(Gp130)将通过鉴定 涉及的顺式代理监管要素及其对应的 转录因子。此外，性激素对人的影响 将研究可溶性IL-6受体的产生和细胞 IL-6、gp80和gp130基因在小鼠骨骼中的体外表达 骨髓培养和未脱钙的性别松质骨切片 类固醇充足和性激素缺乏的小鼠将被识别，使用 原位逆转录聚合酶链式反应结合免疫组织化学方法。此外， IL-6亚家族细胞因子在成骨细胞上调中的作用 在失去性类固醇后在骨髓中形成祖细胞， 以及这些细胞因子和其他生长因子之间的潜在相互作用 对成骨细胞形成的影响因素将进行研究。最后，嵌合体 可溶性IL-6或CNTF受体α的构建，与 识别细胞表面标记的单抗将被用于 选择性地将相应的细胞因子定向到破骨细胞或 成骨细胞前体细胞；并研究这种操作的效果 关于破骨细胞生成和成骨细胞生成。