低流体剪切应力通过 MCAM 和 PECAM-1 细胞粘附分子刺激有害内皮细胞外囊泡的吸收

IF 15.5 1区 医学 Q1 CELL BIOLOGY
Pierre-Michaël Coly, Shruti Chatterjee, Fariza Mezine, Christelle El Jekmek, Cécile Devue, Thomas Nipoti, Stephane Mazlan, Maribel Lara Corona, Florent Dingli, Damarys Loew, Guillaume van Niel, Xavier Loyer, Chantal M. Boulanger
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引用次数: 0

摘要

动脉粥样硬化病变主要形成于暴露于低剪切应力(LSS)的动脉区域,那里的内皮细胞表现出衰老和炎症表型。相反,暴露于高剪切应力(HSS)的区域则受到保护,不会形成斑块。内皮细胞外囊泡(EVs)已被证明能调节炎症和衰老,因此在血管稳态中起着至关重要的作用。虽然之前的研究表明血液动力和EV释放之间存在联系,但剪切应力对内皮EVs释放和吸收的影响仍然难以捉摸。我们的目的是破解暴露在动脉粥样硬化或动脉粥样硬化保护性剪切应力下的内皮细胞中这些过程之间的相互作用。汇合的 HUVEC 暴露于 LSS 或 HSS 24 小时后,通过离心和尺寸排阻色谱法从条件培养基中分离出大的和小的 EVs。通过 TEM、Western 印迹、可调电阻脉冲传感、流式细胞仪和蛋白质组学对它们进行了表征。使用荧光标记的 EVs 进行了吸收实验,并通过流式细胞术和共聚焦显微镜评估了组间差异。我们发现,在 HSS 条件下,条件培养基中大型和小型 EV 的水平分别是 LSS 条件下的 50 倍和 5 倍。体内和体外摄取实验显示,暴露于 LSS 条件下的细胞对 EV 的结合率更高。此外,与 HSS-EVs 或来自血小板、红细胞和白细胞的 EVs 相比,内皮 LSS-EVs 对 HUVECs 的亲和力更大。蛋白质组分析表明,LSS-EV富含粘附蛋白(PECAM1、MCAM),参与了内皮细胞对 EV 的吸收。LSS-EV 还携带线粒体物质,这可能与受体细胞中 ROS 水平的升高有关。这些发现表明,剪切应力影响了EV的生物生成和摄取。鉴于EVs和剪切应力在血管健康中的重要作用,破译这些过程之间的关系可能会产生早期检测和治疗内皮功能障碍的创新策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Low fluid shear stress stimulates the uptake of noxious endothelial extracellular vesicles via MCAM and PECAM-1 cell adhesion molecules

Atherosclerotic lesions mainly form in arterial areas exposed to low shear stress (LSS), where endothelial cells express a senescent and inflammatory phenotype. Conversely, areas exposed to high shear stress (HSS) are protected from plaque development. Endothelial extracellular vesicles (EVs) have been shown to regulate inflammation and senescence, and therefore play a crucial role in vascular homeostasis. Whilst previous studies have shown links between hemodynamic forces and EV release, the effects of shear stress on the release and uptake of endothelial EVs remains elusive. We aim to decipher the interplay between these processes in endothelial cells exposed to atheroprone or atheroprotective shear stress. Confluent HUVECs were exposed to LSS or HSS for 24 h. Large and small EVs were isolated from conditioned medium by centrifugation and size exclusion chromatography. They were characterised by TEM, Western blot, tunable resistive pulse sensing, flow cytometry and proteomics. Uptake experiments were performed using fluorescently-labelled EVs and differences between groups were assessed by flow cytometry and confocal microscopy. We found that levels of large and small EVs in conditioned media were fifty and five times higher in HSS than in LSS conditions, respectively. In vivo and in vitro uptake experiments revealed greater EV incorporation by cells exposed to LSS conditions. Additionally, endothelial LSS-EVs have a greater affinity for HUVECs than HSS-EVs or EVs derived from platelets, erythrocytes and leukocytes. Proteomic analysis revealed that LSS-EVs were enriched in adhesion proteins (PECAM1, MCAM), participating in EV uptake by endothelial cells. LSS-EVs also carried mitochondrial material, which may be implicated in elevating ROS levels in recipient cells. These findings suggest that shear stress influences EV biogenesis and uptake. Given the major role of EVs and shear stress in vascular health, deciphering the relation between these processes may yield innovative strategies for the early detection and treatment of endothelial dysfunction.

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来源期刊
Journal of Extracellular Vesicles
Journal of Extracellular Vesicles Biochemistry, Genetics and Molecular Biology-Cell Biology
CiteScore
27.30
自引率
4.40%
发文量
115
审稿时长
12 weeks
期刊介绍: The Journal of Extracellular Vesicles is an open access research publication that focuses on extracellular vesicles, including microvesicles, exosomes, ectosomes, and apoptotic bodies. It serves as the official journal of the International Society for Extracellular Vesicles and aims to facilitate the exchange of data, ideas, and information pertaining to the chemistry, biology, and applications of extracellular vesicles. The journal covers various aspects such as the cellular and molecular mechanisms of extracellular vesicles biogenesis, technological advancements in their isolation, quantification, and characterization, the role and function of extracellular vesicles in biology, stem cell-derived extracellular vesicles and their biology, as well as the application of extracellular vesicles for pharmacological, immunological, or genetic therapies. The Journal of Extracellular Vesicles is widely recognized and indexed by numerous services, including Biological Abstracts, BIOSIS Previews, Chemical Abstracts Service (CAS), Current Contents/Life Sciences, Directory of Open Access Journals (DOAJ), Journal Citation Reports/Science Edition, Google Scholar, ProQuest Natural Science Collection, ProQuest SciTech Collection, SciTech Premium Collection, PubMed Central/PubMed, Science Citation Index Expanded, ScienceOpen, and Scopus.
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