Koh Eun Jung, Choi Seung Ah, Moon Youn Joo, Lee Heeyoung, Phi Ji Hoon, Kim Seung-Ki
Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital, Seoul, Korea.
Department of Neurosurgery, Seoul National University Hospital, Seoul National University College of Medicine, Seoul, Korea.
Sci Rep. 2025 Jul 9;15(1):24655. doi: 10.1038/s41598-025-08796-4.
Moyamoya disease (MMD) is a chronic cerebrovascular disorder and a leading cause of pediatric stroke. Extracellular vesicles (EVs) carrying microRNAs (miRNAs) play a pivotal role in intercellular communication within cerebrovascular diseases. This study aimed to identify specific miRNAs within plasma-derived EVs from MMD patients and investigate their functional implications. Study subjects included healthy controls (N = 13) and MMD patients (N = 23). EVs were isolated from plasma samples and characterized by transmission electron microscopy, nanoparticle tracking analysis, ExoView, RT-qPCR, and immunoblotting. miRNA profiles were assessed through NanoString analysis. Functional effects of miR-512-3p inhibition were evaluated in MMD endothelial colony-forming cells (ECFCs) by analyzing guanosine triphosphatase (GTPase) activity, tubule formation, and cell viability. MMD-derived EVs exhibited an upregulation of miR-512-3p compared to controls. Bioinformatics analysis identified RHO guanine nucleotide exchange factor 3 (ARHGEF3) as a potential target of miR-512-3p. Inhibition of miR-512-3p in MMD ECFCs resulted in increased expression of ARHGEF3 and its downstream effector RHOA, leading to enhanced GTPase activity and improved tubule formation, indicative of restored angiogenic function. Elevated levels of miR-512-3p within plasma-derived EVs may serve as a novel biomarker for MMD diagnosis. The modulation of ARHGEF3 and subsequent RHOA signaling by targeting miR-512-3p contributes to the dysregulated angiogenesis in MMD.
烟雾病（MMD）是一种慢性脑血管疾病，也是小儿中风的主要原因。携带微小RNA（miRNA）的细胞外囊泡（EV）在脑血管疾病的细胞间通讯中起关键作用。本研究旨在鉴定烟雾病患者血浆来源的细胞外囊泡中的特定微小RNA，并研究其功能意义。研究对象包括健康对照者（N = 13）和烟雾病患者（N = 23）。从血浆样本中分离出细胞外囊泡，并通过透射电子显微镜、纳米颗粒跟踪分析、ExoView、逆转录定量聚合酶链反应（RT-qPCR）和免疫印迹进行表征。通过NanoString分析评估微小RNA谱。通过分析鸟苷三磷酸酶（GTPase）活性、小管形成和细胞活力，评估了miR-512-3p抑制对烟雾病内皮祖细胞集落形成细胞（ECFC）的功能影响。与对照组相比，烟雾病来源的细胞外囊泡表现出miR-512-3p上调。生物信息学分析确定RHO鸟嘌呤核苷酸交换因子3（ARHGEF3）为miR-512-3p的潜在靶标。在烟雾病ECFC中抑制miR-512-3p导致ARHGEF3及其下游效应物RHOA的表达增加，从而导致GTPase活性增强和小管形成改善，表明血管生成功能恢复。血浆来源的细胞外囊泡中miR-512-3p水平升高可能作为烟雾病诊断的新型生物标志物。通过靶向miR-512-3p调节ARHGEF3及随后的RHOA信号传导，导致烟雾病中血管生成失调。
