Department of Experimental Cardiology, University Medical Center Utrecht, Utrecht University, Utrecht, The Netherlands.
Biomolecular Mass Spectrometry and Proteomics, Bijvoet Center for Biomolecular Research and Utrecht Institute for Pharmaceutical Sciences, Utrecht University, Utrecht, The Netherlands.
Commun Biol. 2023 Aug 1;6(1):800. doi: 10.1038/s42003-023-05165-7.
Extracellular vesicles (EVs) are cell-derived lipid bilayer-enclosed particles that play a role in intercellular communication. Cardiac progenitor cell (CPC)-derived EVs have been shown to protect the myocardium against ischemia-reperfusion injury via pro-angiogenic effects. However, the mechanisms underlying CPC-EV-induced angiogenesis remain elusive. Here, we discovered that the ability of CPC-EVs to induce in vitro angiogenesis and to stimulate pro-survival pathways was lost upon EV donor cell exposure to calcium ionophore. Proteomic comparison of active and non-active EV preparations together with phosphoproteomic analysis of activated endothelial cells identified the contribution of candidate protein PAPP-A and the IGF-R signaling pathway in EV-mediated cell activation, which was further validated using in vitro angiogenesis assays. Upon further purification using iodixanol gradient ultracentrifugation, EVs partly lost their activity, suggesting a co-stimulatory role of co-isolated proteins in recipient cell activation. Our increased understanding of the mechanisms of CPC-EV-mediated cell activation will pave the way to more efficient EV-based therapeutics.
细胞外囊泡 (EVs) 是由细胞膜包裹的脂质双层囊泡，在细胞间通讯中发挥作用。心脏祖细胞 (CPC) 衍生的 EVs 通过促血管生成作用被证明可以保护心肌免受缺血再灌注损伤。然而，CPC-EV 诱导血管生成的机制尚不清楚。在这里，我们发现 CPC-EV 诱导体外血管生成和刺激生存相关途径的能力在 EV 供体细胞暴露于钙离子载体时丧失。对活性和非活性 EV 制剂的蛋白质组学比较以及对激活的内皮细胞的磷酸化蛋白质组学分析，确定了候选蛋白 PAPP-A 和 IGF-R 信号通路在 EV 介导的细胞激活中的贡献，这进一步通过体外血管生成实验得到了验证。在用碘克沙醇梯度超速离心进一步纯化后，EV 部分丧失了其活性，这表明共分离的蛋白质在受体细胞激活中具有协同刺激作用。我们对 CPC-EV 介导的细胞激活机制的理解的增加将为更有效的基于 EV 的治疗铺平道路。
