Divisions of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, USA.
Health Sciences Research, Mayo Clinic, Rochester, Minnesota, USA.
Stem Cells Transl Med. 2019 May;8(5):430-440. doi: 10.1002/sctm.18-0171. Epub 2019 Feb 1.
Mesenchymal stem/stromal cells (MSCs) release extracellular vesicles (EVs), which shuttle proteins to recipient cells, promoting cellular repair. We hypothesized that cardiovascular risk factors may alter the pattern of proteins packed within MSC-derived EVs. To test this, we compared the protein cargo of EVs to their parent MSCs in pigs with metabolic syndrome (MetS) and Lean controls. Porcine MSCs were harvested from abdominal fat after 16 weeks of Lean- or MetS-diet (n = 5 each), and their EVs isolated. Following liquid chromatography mass spectrometry proteomic analysis, proteins were classified based on cellular component, molecular function, and protein class. Five candidate proteins were validated by Western blot. Clustering analysis was performed to identify primary functional categories of proteins enriched in or excluded from EVs. Proteomics analysis identified 6,690 and 6,790 distinct proteins in Lean- and MetS-EVs, respectively. Differential expression analysis revealed that 146 proteins were upregulated and 273 downregulated in Lean-EVs versus Lean-MSCs, whereas 787 proteins were upregulated and 185 downregulated in MetS-EVs versus MetS-MSCs. Proteins enriched in both Lean- and MetS-EVs participate in vesicle-mediated transport and cell-to-cell communication. Proteins enriched exclusively in Lean-EVs modulate pathways related to the MSC reparative capacity, including cell proliferation, differentiation, and activation, as well as transforming growth factor-β signaling. Contrarily, proteins enriched only in MetS-EVs are linked to proinflammatory pathways, including acute inflammatory response, leukocyte transendothelial migration, and cytokine production. Coculture with MetS-EVs increased renal tubular cell inflammation. MetS alters the protein cargo of porcine MSC-derived EVs, selectively packaging specific proinflammatory signatures that may impair their ability to repair damaged tissues. Stem Cells Translational Medicine 2019;8:430-440.
间充质干细胞/基质细胞 (MSCs) 释放细胞外囊泡 (EVs)，这些囊泡将蛋白质运送到受体细胞，促进细胞修复。我们假设心血管危险因素可能会改变 MSC 衍生 EV 中包裹的蛋白质模式。为了验证这一点，我们比较了代谢综合征 (MetS) 和瘦型对照猪的 EV 和其亲本 MSC 之间的蛋白质组。猪 MSC 从 16 周的瘦型或 MetS 饮食后的腹部脂肪中收获 (n = 5 个/组)，并分离其 EV。在进行液相色谱-质谱蛋白质组学分析后，根据细胞成分、分子功能和蛋白质类别对蛋白质进行分类。通过 Western blot 验证了 5 种候选蛋白。通过聚类分析确定了富含 EVs 或排除 EVs 的主要功能类别的蛋白质。蛋白质组学分析分别在瘦型和 MetS-EVs 中鉴定出 6690 和 6790 种不同的蛋白质。差异表达分析显示，在瘦型-EVs 中与瘦型-MSCs 相比有 146 个蛋白上调和 273 个蛋白下调，而在 MetS-EVs 中与 MetS-MSCs 相比有 787 个蛋白上调和 185 个蛋白下调。在瘦型和 MetS-EVs 中均富集的蛋白质参与囊泡介导的运输和细胞间通讯。仅在瘦型-EVs 中富集的蛋白质调节与 MSC 修复能力相关的途径，包括细胞增殖、分化和激活，以及转化生长因子-β信号通路。相反，仅在 MetS-EVs 中富集的蛋白质与促炎途径相关，包括急性炎症反应、白细胞穿越内皮迁移和细胞因子产生。与 MetS-EVs 共培养增加了肾小管细胞的炎症。MetS 改变了猪 MSC 衍生 EV 的蛋白质组，选择性地包装特定的促炎特征，这可能会损害它们修复受损组织的能力。干细胞转化医学 2019;8:430-440.
