Differential expression of circular RNAs in bone marrow-derived exosomes from essential thrombocythemia patients
Differential expression of circular RNAs in bone marrow-derived exosomes from essential thrombocythemia patients
复制标题
原发性血小板增多症患者骨髓来源的外泌体中环状RNA的差异表达
DOI：
10.1002/cbin.11534
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发表时间：
2021
影响因子：
3.9
通讯作者：
Xu Dan
中科院分区：
文献类型：
--
作者：
Wang Qiang;Yu Guopan;He Han;Zheng Zhongxin;Li Xuan;Lin Ren;Xu Dan
环状RNA（circRNA）与多种血液病的发病机制密切相关。然而，人们对 circRNA 在原发性血小板增多症 (ET) 发展中的潜在功能知之甚少。本研究主要研究 ET 患者骨髓中 circRNA 谱的变化。通过纳米颗粒跟踪分析（NTA）方法验证了源自人骨髓组织的外泌体的大小。通过蛋白质印迹分析分析外泌体中 CD63 和 TSG101 的表达。通过高通量测序对骨源性外泌体中 circRNA 的分布和差异表达进行了表征。在此，分别通过桑格测序和实时聚合酶链反应验证了circRNA的环状结构和表达。使用 Cytoscape 软件预测 circRNA-miRNA-mRNA 网络。我们检测了circ_0014614对K562细胞向巨核细胞转化的影响。来自 ET 患者和健康志愿者骨髓的外泌体直径在 70 至 140 nm 之间，并表达高 CD63 和 TSG101。同时，ET患者骨髓源性外泌体中的circRNA谱发生显着改变，其中circDAP3、circASXL1和circRUNX1在ET患者中显着下调，从而为circRNA在ET发病机制中的作用提供了新的见解。除此之外，这三个circRNA靶向的circRNA编码基因和miRNA-mRNA网络参与了各种生物过程和信号通路。 circ_0014614可以抑制K562细胞向巨核细胞的分化。对这三种差异表达的 circRNA 的潜在功能及其与特定 miRNA 的相互作用的预测可以为基于 circRNA 的 ET 诊断和治疗提供基础。
Circular RNAs (circRNA) are closely associated with the pathogenesis of various hematological diseases. However, little is known about the potential functions of circRNAs in essential thrombocythemia (ET) development. The circRNA profile alterations in the bone marrow of ET patients were mainly investigated in this study. The sizes of exosomes derived from human bone marrow tissues were validated by the nanoparticle tracking analysis (NTA) method. CD63 and TSG101 expressions in exosomes were analyzed by western blot analysis. The profiles and differential expression of circRNAs in bone‐derived exosomes were characterized by high‐throughput sequencing. Herein, circular structures and expression of circRNAs were verified by Sanger sequencing and real‐time polymerase chain reaction, respectively. The circRNA‐miRNA‐mRNA networks were predicted using the Cytoscape software. And we detected the effect of circ_0014614 on the transformation of K562 cells into megakaryocytes. Exosomes derived from the bone marrow of ET patients and healthy volunteers showed a diameter between 70 and 140 nm and expressed high CD63 and TSG101. Meanwhile, the circRNA profiles were significantly altered in bone marrow–derived exosomes from ET patients, among which circDAP3, circASXL1, and circRUNX1 were significantly downregulated in ET patients, thus conferring a new insight into the role of circRNAs in the pathogenesis of ET. Besides this, circRNA‐encoding genes and miRNA‐mRNA networks targeted by this three circRNA were involved in various biological processes and signaling pathways. And circ_0014614 could inhibit K562 cells' differentiation into megakaryocytes. The predictions of the potential function of these three differentially expressed circRNAs along with their interaction with specific miRNAs could provide a basis for circRNA‐based ET diagnosis and treatment.