Proteome-wide Expression Study of Osteogenic Cells

骨质疏松症是一个主要的公共健康问题，特别是在女性中。骨质疏松症是其主要特点 矿物质密度(BMD)。BMD有60%的高遗传度。女性的骨密度比男性低得多。我们和其他人 证明了一些BMD基因/基因组区域具有性别特异性。更年期是最重要的 女性生活中的生理事件，与女性特有的快速骨质流失有关。 骨髓间充质干细胞(BMMSCs)和外周血单核细胞(PBM)是 成骨细胞(骨形成细胞)和破骨细胞(骨吸收细胞)。 我们的假设是，女性BMMSCs和PBM中蛋白质表达谱的变化是基础 女性BMD变异的机制及其与更年期的关系。 我们的主要目标是确定在女性BMMSCs和PBM中差异表达的蛋白质：1) 高BMD与低BMD；2)绝经前后，从而识别与以下因素相关的蛋白质(及其基因) 骨髓间充质干细胞和外周血中的女性骨质疏松症和绝经。在女性身上发现的有意义的蛋白质将是 在男性样本中进行了检测。 连同项目2，我们将招募80名其他方面健康的女性和80名年龄匹配的其他方面健康的女性 年龄50-55岁的男性，包括40名低BMD和40名高BMD(年龄匹配的人群底部或顶部20% 每个性别，都是高加索人。每个女性BMD组包括20岁之前和20岁之前匹配的女性 绝经后的女性。我们将采集新鲜的骨髓(我们已经对其进行了广泛的已发表研究 经验)和每个受试者的外周血液样本。骨髓间充质干细胞和骨髓间充质干细胞将被隔离并同等 分成两等份，一份用于项目2，一份用于项目3。在这个项目3中，我们将提取总蛋白质 从分离的BMMSCs和PBM中分离出等量的细胞。蛋白质组学实验和分析将在 女性使用MD-Nano-LC-MS/MS发现的显著差异表达的蛋白质将通过 用雌性样本进行蛋白质印迹，以确定其在雌性中的重要性。在女性体内被证实的重要蛋白质将 通过Western blotting对男性样本进行性别特异性检查。 在高加索人中获得的结果将在中国样本中交叉核对种族共性/特异性。 已鉴定的蛋白质及其基因的分子和细胞功能研究将作为例证继续进行。 在本SCOR的项目2中。 这项研究的主要结果(特别是从PBM获得的结果)可用于设计习惯 诊断女性骨质疏松症预后的蛋白质抗体芯片和/或蛋白质标记物。尤其是， 与中的DMA多态数据结合时，结果将用于提供一些功能证据 项目1和本SCOR项目2中的DMA微阵列数据，以强大而高效地识别基因和一些 它们对女性骨质疏松症的作用。

Osteoporosis is a major public health problem, especially in women. It is mainly characterized by low bone mineral density (BMD). BMD has high heritability of > 60%. Women have much lower BMD than men. We and others demonstrated that some BMD genes/genomic regions are gender specific. Menopause is a most significant physiology event in female's life and is associated with female-specific rapid bone loss. Bone marrow mesenchymal stem cells (BMMSCs) and peripheral blood monocytes (PBMs), are precursors for osteoblasts (bone formation cells) and osteoclasts (bone resorption cells), respectively. Our hypothesis is that changes in the protein expression profiles in female BMMSCs and PBMs underlie mechanisms of female BMD variation and are associated with menopause. Our major goals here are to identify proteins differentially expressed in BMMSCs and PBMs in women: 1) with high vs. low BMD; 2) before and after menopause, and thus identify proteins (and their genes) associated with female osteoporosis and menopause in BMMSCs and PBMs. The proteins identified significant in females will be examined in male samples. Together with Project 2, we will recruit 80 otherwise healthy females and 80 age-matched otherwise healthy males aged 50-55, including 40 subjects with low and 40 with high BMD (age matched population bottom or top 20% respectively) for each sex, all Caucasian. Each female BMD group includes 20 pre- and 20 age matched post-menopausal women. We will take fresh bone marrow (for which we have had extensive published research experience) and peripheral blood samples from each subject. BMMSCs and PBMs will be isolated and equally divided into two aliquots, one for Project 2 and one for Project 3. In this Project 3, we will extract the total proteins from aliquots of isolated BMMSCs and PBMs. Proteomic profiling experiments and analyses will be performed on Females using MD-nano-LC-MS/MS. Significant differentially expressed proteins identified will be verified by Western blotting with female samples for their importance in females. Significant proteins verified in females will be examined by Western blotting with male samples for sex-specificity. The results obtained in Caucasians will be cross checked for ethnic generality/specificity in Chinese samples. The molecular and cellular functional studies of the identified proteins and their genes will be pursued as exemplified in Project 2 of this SCOR. The major results (particularly those obtained from PBMs) of this study may be used to design customary diagnostic protein antibody chips and/or protein markers for prognosis of female osteoporosis. In particular, the results will be used to provide some functional evidence, when combined with the DMA polymorphism data in Project 1 and DMA microarray data in Project 2 of this SCOR to powerfully and efficiently identify genes and some of their functions for female osteoporosis.