MicroRNA-183-5p Increases with Age in Bone-Derived Extracellular Vesicles, Suppresses Bone Marrow Stromal (Stem) Cell Proliferation, and Induces Stem Cell Senescence
MicroRNA-183-5p Increases with Age in Bone-Derived Extracellular Vesicles, Suppresses Bone Marrow Stromal (Stem) Cell Proliferation, and Induces Stem Cell Senescence
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DOI：
10.1089/ten.tea.2016.0525
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发表时间：
2017-11-01
影响因子：
4.1
通讯作者：
Hamrick, Mark W.
中科院分区：
文献类型：
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作者：
Davis, Colleen;Dukes, Amy;Hamrick, Mark W.
微囊泡和外切体介导的microRNAs(MiRNAs)是细胞与细胞间通讯的一种新的细胞和分子途径。在这项研究中，我们验证了这样一种假设，即这些细胞外小泡(EV)及其miRNAs可能会随着年龄的变化而变化，从而导致与年龄相关的干细胞功能障碍。从幼龄(3~4月龄)和老龄(24~28月龄)小鼠骨髓间质液(上清液)中分离EVS，以确定EVS的大小、浓度和miRNA在体内是否随年龄变化而改变。结果表明，从小鼠骨髓分离的EVS呈CD63和CD9阳性，幼年和老年小鼠骨髓EVS的浓度和大小分布相似。BioAnalyzer数据显示，年轻和老年小鼠的EVS都高度富含miRNAs，并且年轻和老年小鼠的骨髓EVS的miRNA谱明显不同。具体地说，miR-183簇(miR-96/-182/-183)在老年EV中高表达。体外实验表明，老年EV被原代骨髓基质细胞(BMSCs)内吞，这些老年EV抑制年轻BMSCs的成骨分化。将miR-183-5p模拟物导入骨髓间充质干细胞，可减少细胞增殖和成骨分化，增加衰老，并降低靶基因miR-183-5p的血红素氧合酶-1(Hmox1)的蛋白水平。体外利用H_2O_2诱导氧化应激实验表明，H_2O_2处理的BMSCs增加了BMSC来源的EV中miR-183-5P的丰度，Amplex Red实验表明H_2O_2在骨髓微环境中随着年龄的增长而增加。综上所述，这些数据表明，衰老和氧化应激可以显著改变EV在骨髓微环境中的miRNA载量，这可能反过来通过降低Hmox1的活性在干细胞衰老和成骨分化中发挥作用。
Microvesicle-and exosome-mediated transport of microRNAs (miRNAs) represents a novel cellular and molecular pathway for cell-cell communication. In this study, we tested the hypothesis that these extracellular vesicles (EVs) and their miRNAs might change with age, contributing to age-related stem cell dysfunction. EVs were isolated from the bone marrow interstitial fluid (supernatant) of young (3-4 months) and aged (24-28 months) mice to determine whether the size, concentration, and miRNA profile of EVs were altered with age in vivo. Results show that EVs isolated from bone marrow are CD63 and CD9 positive, and the concentration and size distribution of bone marrow EVs are similar between the young and aged mice. Bioanalyzer data indicate that EVs from both young and aged mice are highly enriched in miRNAs, and the miRNA profile of bone marrow EVs differs significantly between the young and aged mice. Specifically, the miR-183 cluster (miR-96/-182/-183) is highly expressed in aged EVs. In vitro assays demonstrate that aged EVs are endocytosed by primary bone marrow stromal cells (BMSCs), and these aged EVs inhibit the osteogenic differentiation of young BMSCs. Transfection of BMSCs with miR-183-5p mimic reduces cell proliferation and osteogenic differentiation, increases senescence, and decreases protein levels of the miR-183-5p target heme oxygenase-1 (Hmox1). In vitro assays utilizing H2O2-induced oxidative stress show that H2O2 treatment of BMSCs increases the abundance of miR-183-5p in BMSC-derived EVs, and Amplex Red assays demonstrate that H2O2 is elevated in the bone marrow microenvironment with age. Together, these data indicate that aging and oxidative stress can significantly alter the miRNA cargo of EVs in the bone marrow microenvironment, which may in turn play a role in stem cell senescence and osteogenic differentiation by reducing Hmox1 activity.