Proteomics reveals multiple routes to the osteogenic phenotype in mesenchymal stem cells
Proteomics reveals multiple routes to the osteogenic phenotype in mesenchymal stem cells
复制标题
DOI：
10.1186/1471-2164-8-380
复制
发表时间：
2007-10-19
期刊：
影响因子：
4.4
通讯作者：
Plopper, George E.
中科院分区：
文献类型：
--
作者：
Bennett, Kristin P.;Bergeron, Charles;Plopper, George E.
背景：近期，我们证明了用地塞米松刺激的人间充质干细胞（hMSC）在成骨分化过程中会发生基因聚焦（《干细胞发育》14(6): 1608 - 20, 2005）。在此，我们检测了另外三群hMSC在通过与促成骨细胞外基质（ECM）蛋白（胶原蛋白I、玻连蛋白或层粘连蛋白 - 5）接触或成骨培养基补充剂（OS培养基）刺激进行成骨分化时的蛋白质表达谱。具体而言，我们用已知的基因本体对这四种蛋白质表达谱以及来自未处理的hMSC和分化的人成骨细胞（hOST）的表达谱进行注释，并将它们作为一个张量进行分析，其模式包括表达的蛋白质、基因本体和刺激物。
结果：在基因本体空间进行的直接成分分析确定了三个成分，它们解释了hMSC、成骨细胞以及四群受刺激的hMSC之间90%的差异。定向成分沿着由hMSC和hOST表达谱差异所创建的分化轴绘制受刺激干细胞群的分化阶段。令人惊讶的是，用ECM蛋白处理的hMSC比用OS培养基处理的hMSC更接近成骨细胞。此外，第二个成分表明胶原蛋白I和玻连蛋白刺激的hMSC的蛋白质组图谱与OS刺激的细胞不同。三模式张量分析揭示了对于表征未处理的hMSC、部分分化的hMSC和hOST之间表型变异至关重要的其他关键蛋白质。
结论：OS刺激的hMSC和ECM - hMSC的蛋白质组图谱之间的差异表征了在成为成骨细胞过程中的不同过渡表型。这一结论是通过三模式张量分析得出的，并使用接种在层粘连蛋白 - 5上的hMSC进行了验证。
Background: Recently, we demonstrated that human mesenchymal stem cells (hMSC) stimulated with dexamethazone undergo gene focusing during osteogenic differentiation (Stem Cells Dev 14( 6): 1608-20, 2005). Here, we examine the protein expression profiles of three additional populations of hMSC stimulated to undergo osteogenic differentiation via either contact with pro-osteogenic extracellular matrix (ECM) proteins (collagen I, vitronectin, or laminin-5) or osteogenic media supplements (OS media). Specifically, we annotate these four protein expression profiles, as well as profiles from nave hMSC and differentiated human osteoblasts (hOST), with known gene ontologies and analyze them as a tensor with modes for the expressed proteins, gene ontologies, and stimulants.Results: Direct component analysis in the gene ontology space identifies three components that account for 90% of the variance between hMSC, osteoblasts, and the four stimulated hMSC populations. The directed component maps the differentiation stages of the stimulated stem cell populations along the differentiation axis created by the difference in the expression profiles of hMSC and hOST. Surprisingly, hMSC treated with ECM proteins lie closer to osteoblasts than do hMSC treated with OS media. Additionally, the second component demonstrates that proteomic profiles of collagen I- and vitronectin-stimulated hMSC are distinct from those of OS-stimulated cells. A three-mode tensor analysis reveals additional focus proteins critical for characterizing the phenotypic variations between nave hMSC, partially differentiated hMSC, and hOST.Conclusion: The differences between the proteomic profiles of OS-stimulated hMSC and ECM-hMSC characterize different transitional phenotypes en route to becoming osteoblasts. This conclusion is arrived at via a three-mode tensor analysis validated using hMSC plated on laminin-5.