Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB-CSIC), Campus Universidad Autónoma de Madrid, 28049, Madrid, Spain.
Cardiovascular Development and Repair Department, Spanish National Cardiovascular Research Center (CNIC), Melchor Fernández Almagro 3, 28029, Madrid, Spain.
Sci Rep. 2019 Mar 15;9(1):4647. doi: 10.1038/s41598-019-39571-x.
Adult cardiac progenitor/stem cells (CPC/CSC) are multipotent resident populations involved in cardiac homeostasis and heart repair. Assisted by complementary RNAseq analysis, we defined the fraction of the CPC proteome associable with specific functions by comparison with human bone marrow mesenchymal stem cells (MSC), the reference population for cell therapy, and human dermal fibroblasts (HDF), as a distant reference. Label-free proteomic analysis identified 526 proteins expressed differentially in CPC. iTRAQ analysis confirmed differential expression of a substantial proportion of those proteins in CPC relative to MSC, and systems biology analysis defined a clear overrepresentation of several categories related to enhanced angiogenic potential. The CPC plasma membrane compartment comprised 1,595 proteins, including a minimal signature of 167 proteins preferentially or exclusively expressed by CPC. CDH5 (VE-cadherin), OX2G (OX-2 membrane glycoprotein; CD200), GPR4 (G protein-coupled receptor 4), CACNG7 (calcium voltage-gated channel auxiliary subunit gamma 7) and F11R (F11 receptor; junctional adhesion molecule A; JAM-A; CD321) were selected for validation. Their differential expression was confirmed both in expanded CPC batches and in early stages of isolation, particularly when compared against cardiac fibroblasts. Among them, GPR4 demonstrated the highest discrimination capacity between all cell lineages analyzed.
成人心肌祖细胞/干细胞（CPC/CSC）是多能的常驻细胞群，参与心脏内稳态和心脏修复。通过互补的 RNAseq 分析，我们将 CPC 蛋白质组中与特定功能相关的部分与细胞治疗的参考人群——人骨髓间充质干细胞（MSC）和作为遥远参照的人真皮成纤维细胞（HDF）进行比较，从而对其进行了定义。无标记蛋白质组学分析鉴定出 526 种在 CPC 中差异表达的蛋白质。iTRAQ 分析证实了这些蛋白质中有相当一部分在 CPC 中相对于 MSC 表达存在差异，系统生物学分析定义了几个与增强血管生成潜力相关的类别明显过表达。CPC 的质膜区室包含 1595 种蛋白质，其中包括 167 种 CPC 特异性或优先表达的最小蛋白质特征。CDH5（VE-钙粘蛋白）、OX2G（OX-2 膜糖蛋白；CD200）、GPR4（G 蛋白偶联受体 4）、CACNG7（钙电压门控通道辅助亚基 gamma 7）和 F11R（F11 受体；连接黏附分子 A；JAM-A；CD321）被选中进行验证。在扩增的 CPC 批次和早期分离阶段都证实了它们的差异表达，特别是与心肌成纤维细胞相比。其中，GPR4 在分析的所有细胞谱系之间表现出最高的区分能力。
