Combined Analysis of Surface Protein Profile and microRNA Expression Profile of Exosomes Derived from Brain Microvascular Endothelial Cells in Early Cerebral Ischemia.
Combined Analysis of Surface Protein Profile and microRNA Expression Profile of Exosomes Derived from Brain Microvascular Endothelial Cells in Early Cerebral Ischemia.
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早期脑缺血时脑微血管内皮细胞来源的外泌体表面蛋白谱和 microRNA 表达谱的联合分析
DOI：
10.1021/acsomega.1c03248
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发表时间：
2021-08-31
期刊：
影响因子：
4.1
通讯作者：
Liu W
中科院分区：
文献类型：
--
作者：
Yang D;Li Z;Gao G;Li X;Liao Z;Wang Y;Li W;Zhang Y;Liu W
内皮细胞损伤是急性缺血性脑卒中恶化的重要病理基础。我们前期的研究主要是探讨脑缺血早期血脑屏障（blood-brain barrier，BBB）内皮细胞损伤的机制。外泌体在神经血管通讯中起重要作用。然而，来源于BBB内皮细胞的外泌体在早期缺血性卒中中的特征知之甚少。我们将培养的脑微血管内皮细胞（bEnd.3）暴露于3 h氧葡萄糖剥夺（OGD）以模拟体外早期脑缺血，并通过miRNA测序和邻近条形码分析（PBA）比较来自bEnd.3细胞的外泌体的miRNA和表面蛋白含量。在暴露于OGD 3 h后，通过miRNA-Seq在bEnd. 3细胞中鉴定了总共346种差异miRNA（159种上调和187种下调）。此外，基因本体（GO）和KEGG通路分析表明，细胞增殖和血管生成相关的miRNA受到显着影响。前8个miRNAs的异常变化通过定量聚合酶链反应（qPCR）进一步验证。PBA实验表明，在缺血条件下，携带以下蛋白的外泌体的数量显著增加，包括bFGF、CD 146、EPHA 2、ABCB 5和ITGB 2。这些蛋白质与血管生成、细胞增殖和细胞炎症有关。结合PBA数据和miRNA-Seq数据的网络分析显示，79个miRNA与24个膜蛋白相关，并预测存在与多种miRNA分子相关的表面蛋白，如ITGA 9，XIAP，ADAM 1，ITGA 2，ITGA 3，PDPN和ITGB 1。同时，存在与各种表面蛋白相关的miRNA，包括miR-410- 3 p、miR-378 b和miR-1960。综上所述，我们的数据首次证明了缺血性微血管内皮细胞来源的外泌体miRNAs和表面蛋白谱的变化，这可能为缺血性脑卒中的BBB保护提供新的治疗靶点。
Endothelial cell damage is an important pathological basis for the deterioration of acute ischemia stroke. Our previous studies have been exploring the mechanism of blood–brain barrier (BBB) endothelial cell injury in the early stage of cerebral ischemia. Exosomes act as an important intercellular player in neurovascular communication. However, the characteristic of exosomes derived from BBB endothelial cells in early ischemic stroke is poorly understood. We exposed cultured brain microvascular endothelial cells (bEnd.3) to 3 h oxygen glucose deprivation (OGD) to mimic early cerebral ischemia in vitro and compared miRome and surface protein contents of exosomes derived from bEnd.3 cells by miRNA sequencing and the proximity barcoding assay (PBA). A total of 346 differentially miRNA (159 upregulated and 187 downregulated) were identified via miRNA-Seq in bEnd.3 cells after exposure to OGD for 3 h. Moreover, Gene Ontology (GO) and KEGG pathway analyses showed that cell proliferation- and angiogenesis-associated miRNAs were significantly affected. The abnormal changes in top eight miRNAs were further verified by a quantitative polymerase chain reaction (qPCR). PBA experiments showed that the numbers of exosomes carrying the following proteins increased significantly under ischemia, including bFGF, CD146, EPHA2, ABCB5, and ITGB2. These proteins were related to angiogenesis, cell proliferation, and cell inflammation. The network analysis combining PBA data with miRNA-Seq data showed that 79 miRNAs were related to 24 membrane proteins and predicted that there were surface proteins associated with a variety of miRNA molecules, such as ITGA9, XIAP, ADAM1, ITGA2, ITGA3, PDPN, and ITGB1. Meanwhile, there were miRNAs related to various surface proteins including miR-410-3p, miR-378b, and miR-1960. Taken together, our data demonstrated for the first time the changes of exosomal miRNAs and surface protein profiles derived from ischemic microvascular endothelial cells, which may provide new therapeutic targets for BBB protection in ischemic stroke.
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DOI：
10.1126/science.aau6977
发表时间：
2020-02-07
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Kalluri R;LeBleu VS
通讯作者：
LeBleu VS
影响因子：
7.3
作者：
Haqqani AS;Delaney CE;Tremblay TL;Sodja C;Sandhu JK;Stanimirovic DB
通讯作者：
Stanimirovic DB
DOI：
10.26355/eurrev_201911_19536
发表时间：
2019-01-01
影响因子：
3.3
作者：
Li, Z-Y.;Long, Q-F.;Zhang, W-H.
通讯作者：
Zhang, W-H.
影响因子：
5.5
作者：
Abdullahi, Wazir;Tripathi, Dinesh;Ronaldson, Patrick T.
通讯作者：
Ronaldson, Patrick T.
影响因子：
6.3
作者：
Krueger, Martin;Bechmann, Ingo;Michalski, Dominik
通讯作者：
Michalski, Dominik