Exosomal MicroRNA Transfer Into Macrophages Mediates Cellular Postconditioning.
Exosomal MicroRNA Transfer Into Macrophages Mediates Cellular Postconditioning.
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DOI：
10.1161/circulationaha.116.024590
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发表时间：
2017-07-11
期刊：
影响因子：
37.8
通讯作者：
Marbán E
中科院分区：
文献类型：
--
作者：
de Couto G;Gallet R;Cambier L;Jaghatspanyan E;Makkar N;Dawkins JF;Berman BP;Marbán E
巨噬细胞来源的细胞（CDCs）通过独特的巨噬细胞（M β）极化在急性心肌梗死（MI）中赋予心脏保护作用。在这里，我们证明了CDC分泌的外泌体（CDCexo）概括了CDC治疗的心脏保护作用，称为细胞后处理。在冠状动脉内输注CDCexo、惰性成纤维细胞外泌体（Fbexo;对照）或媒介物之前，大鼠和猪经历由缺血-再灌注诱导的MI。两天后，量化梗死面积。从心脏组织或骨髓中分离巨噬细胞用于下游分析。RNA测序用于确定外泌体含量和M β中基因表达谱的改变。在大鼠和猪MI模型中，再灌注后给予CDCexo而非Fbexo可减少梗死面积。此外，CDCexo减少梗死组织中的CD 68 + M β的数量，并改变M β的极化状态，从而模拟CDC诱导的极化状态。相对于Fbexo，CDCexo富含几种miRNA（包括miR-146 a、miR-181 b和miR-126）。对CDCexo引发的M β的全转录组数据的反向途径分析表明，miR-181 b是CDC诱导的M β极化的重要（p=1.3×10−21）候选介质，蛋白激酶C δ（PKCδ）是下游靶点。选择性负载miR-181 b的顺时针惰性Fbexo改变MI大鼠模型中的MI表型并赋予心脏保护功效PKCδ-抑制的M β的连续转移再现了心脏保护作用。我们的数据支持这样的假设，即miR-181 b从CDC到M β的外泌体转移降低了PKCδ转录水平，并成为再灌注后给予CDC的心脏保护作用的基础。
Cardiosphere-derived cells (CDCs) confer cardioprotection in acute myocardial infarction (MI) via distinctive macrophage (Mϕ) polarization. Here we demonstrate that CDC-secreted exosomes (CDCexo) recapitulate the cardioprotective effects of CDC therapy known as cellular postconditioning. Rats and pigs underwent MI induced by ischemia-reperfusion prior to intracoronary infusion of CDCexo, inert fibroblast exosomes (Fbexo; control), or vehicle. Two days later, infarct size was quantified. Macrophages were isolated from cardiac tissue or bone marrow for downstream analyses. RNA-sequencing was used to determine exosome content and alterations in gene expression profiles in Mϕ. Administration of CDCexo, but not Fbexo, after reperfusion reduces infarct size in rat and pig models of MI. Furthermore, CDCexo reduce the number of CD68+ Mϕ within infarcted tissue and modify the polarization state of Mϕ so as to mimic that induced by CDCs. CDCexo are enriched in several miRNAs (including miR-146a, miR-181b, and miR-126) relative to Fbexo. Reverse pathway analysis of whole-transcriptome data from CDCexo-primed Mϕ implicated miR-181b as a significant (p=1.3×10−21) candidate mediator of CDC-induced Mϕ polarization and protein kinase C δ (PKCδ) as a downstream target. Otherwise-inert Fbexo loaded selectively with miR-181b alter Mϕ phenotype and confer cardioprotective efficacy in a rat model of MI. Adoptive transfer of PKCδ-suppressed Mϕ recapitulates cardioprotection. Our data support the hypothesis that exosomal transfer of miR-181b from CDCs into Mϕ reduces PKCδ transcript levels and underlies the cardioprotective effects of CDCs administered after reperfusion.