CYTOKINE SIGNAL TRANSDUCTION IN OSTEOBLASTIC CELLS

成骨细胞中的细胞因子信号转导

• 批准号: 6374333
• 负责人: Teresita M. Bellido
• 金额: $ 7.42万
• 依托单位: UNIV OF ARKANSAS FOR MED SCIS
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-15 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6374333
• 关键词: 1,25 dihydroxycholecalciferol; JAK kinase; RNase protection assay; SDS polyacrylamide gel electrophoresis; apoptosis; biological signal transduction; cell differentiation; cell growth regulation; cytogenetics; cytokine receptors; gel mobility shift assay; genetic regulation; glucocorticoids; green fluorescent proteins; hormone regulation /control mechanism; interleukin 6; laboratory mouse; osteoblasts; parathyroid hormones; phosphorylation; polymerase chain reaction; receptor binding; receptor expression; retinoids; sex hormones; western blottings

This is an application for an Independent Scientist Award (ISA) K02. The applicant is a very productive, experienced, and NIH- funded investigator. Her research focuses on cytokine signal transduction in osteoblastic cells and its effects on bone remodeling. The applicant works in a rich and productive environment provided by the Center for Osteoporosis and Metabolic Bone Diseases at the University of Arkansas for Med. Sciences. Her work as an independent investigator demonstrated that osteoblastic cells display receptors for IL-6 type cytokines (IL- 6, IL-11, LIF, CNTF, and OSM); that receptor activation induces differentiation and prevents apoptosis of osteoblastic cells; and that, in an osteoblastic cell line, cytokine effects require the transcriptional activation of the cyclin dependent kinase inhibitor p21WAF1, SDI1, CIP1. In addition, cells representing different stages of differentiation express different receptor repertoire, and receptor expression is modulated by systemic hormones. Based on this, she proposes the following interrelated working hypotheses: IL-6 type cytokines promote differentiation and prevent apoptosis of osteoblastic cells by regulating the expression of p21WAF1, SDI1, CIP1. Different receptor repertoires are expressed at different stages of osteoblast differentiation in vivo, and systemic hormones influence the differentiating and anti-apoptotic effects of the cytokines by modulating receptor expression. To test these hypotheses, she proposes to: 1) Investigate the mechanism of the anti-apoptotic effect of the cytokines using MG-63 osteoblastic cells and p21 deficient osteoblasts, and transfections with dominant negative and constitutively or inducibly active molecules of the cytokine signaling pathway. 2) Determine the effects of systemic hormones on receptor expression; and examine whether these changes alter cell responses to the cytokines. And, 3) Determine the pattern of distribution of cytokine receptors in osteoblastic cells in vivo, in murine bone marrow cell aspirates and sections of undecalcified bone from mice, by in situ RT-PCR in combination with histostaining and dynamic bone histomorphometry. The K02 award will permit the applicant to focus her research on cytokine signaling in bone and to enhance her cell/molecular biology skills, as the immediate goals; and to achieve scientific maturity in a uniquely strong environment as her academic career progresses.

这是独立科学家奖 (ISA) K02 的申请。 申请人是一位非常富有成效、经验丰富且由 NIH 资助的研究者。 她的研究重点是成骨细胞中的细胞因子信号转导及其对骨重塑的影响。 申请人在阿肯色大学医学分校骨质疏松症和代谢性骨疾病中心提供的丰富且富有成效的环境中工作。科学。她作为独立研究者的工作表明，成骨细胞展示 IL-6 型细胞因子（IL-6、IL-11、LIF、CNTF 和 OSM）的受体；受体激活诱导分化并防止成骨细胞凋亡；在成骨细胞系中，细胞因子效应需要细胞周期蛋白依赖性激酶抑制剂 p21WAF1、SDI1、CIP1 的转录激活。 此外，代表不同分化阶段的细胞表达不同的受体库，并且受体表达受到全身激素的调节。 基于此，她提出以下相互关联的工作假设：IL-6型细胞因子通过调节p21WAF1、SDI1、CIP1的表达促进成骨细胞分化并防止凋亡。 体内成骨细胞分化的不同阶段表达不同的受体库，全身激素通过调节受体表达影响细胞因子的分化和抗凋亡作用。 为了检验这些假设，她建议：1）利用MG-63成骨细胞和p21缺陷型成骨细胞，以及细胞因子信号通路显性失活和组成型或诱导型活性分子的转染，研究细胞因子的抗凋亡作用机制。 2）确定全身激素对受体表达的影响；并检查这些变化是否改变细胞对细胞因子的反应。 并且，3) 通过原位 RT-PCR 结合组织染色和动态骨组织形态计量学，确定体内成骨细胞、小鼠骨髓细胞抽吸物和小鼠未脱钙骨切片中细胞因子受体的分布模式。 K02 奖将允许申请人将研究重点放在骨细胞因子信号传导上，并提高其细胞/分子生物学技能，作为近期目标；随着她学术生涯的进步，在独特的强大环境中实现科学成熟。