Zhao Ze-Liang, Liu Chao, Wang Qi-Zhang, Wu Hai-Wei, Zheng Jia-Wei
Department of Oromaxillofacial Head and Neck Oncology, Shanghai Ninth People's Hospital, College of Stomatology, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Oral and Maxillofacial Surgery, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.
Ann Transl Med. 2022 Jun;10(11):621. doi: 10.21037/atm-21-4138.
Infantile hemangioma (IH) is the most common benign vascular tumor of infancy and is proposed to arise from hemangioma stem cells (HemSCs). Therapies for IH include oral beta-blockers, surgery, and the delivery of novel therapeutic agents, such as bioactive microRNAs (miRNAs). However, in the extracellular environment, miRNA is easily hydrolyzed by RNase. miR-187-3p has previously been confirmed to promote or inhibit various malignancies, but its role in the development and progression of IH remains unclear.
In this study, engineered exosomes (E-exos) were exploited to deliver miR-187-3p into HemSCs. The E-exos were generated by introducing miR-187-3p mimics into human adipose mesenchymal stem cell-derived exosomes (hAMSC-exos) via electroporation. The expression and secretion of miR-187-3p were examined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Western blot analysis, transmission electron microscopy (TEM), and dynamic light scattering (DLS) were used to characterize the exosomes. The effects of the E-exos on HemSC viability were examined using the tube formation assay and the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay. Western blot analysis was used to evaluate the effects of E-exos on Notch-1, Notch-4, and Jagged-1 expression in HemSCs.
E-exos did not differ significantly from hAMSC-exos in terms of morphology, particle size, or surface markers. E-exos could be internalized by HemSCs, and the course of cellular uptake of E-exos was time dependent. After 12 hours of treatment, E-exos significant inhibited tube formation. Notch signaling was also inhibited by miR-187-3p loading by E-exos. E-exos showed excellent inhibitory effects against HemSC proliferation via Notch signaling.
This study provides a foundation for using hAMSC-exos to optimize current clinical options to facilitate IH treatment and deliver therapeutic agents in the future.
婴儿血管瘤（IH）是婴儿期最常见的良性血管肿瘤，被认为起源于血管瘤干细胞（HemSCs）。IH的治疗方法包括口服β受体阻滞剂、手术以及新型治疗药物的递送，如生物活性微小RNA（miRNAs）。然而，在细胞外环境中，miRNA很容易被核糖核酸酶水解。miR-187-3p先前已被证实可促进或抑制各种恶性肿瘤，但其在IH发生发展中的作用仍不清楚。
在本研究中，利用工程化外泌体（E-exos）将miR-187-3p递送至HemSCs。通过电穿孔将miR-187-3p模拟物导入人脂肪间充质干细胞来源的外泌体（hAMSC-exos）中，从而产生E-exos。通过定量逆转录-聚合酶链反应（qRT-PCR）检测miR-187-3p的表达和分泌情况。采用蛋白质免疫印迹分析、透射电子显微镜（TEM）和动态光散射（DLS）对外泌体进行表征。使用管腔形成试验和3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四氮唑溴盐试验检测E-exos对HemSC活力的影响。采用蛋白质免疫印迹分析评估E-exos对HemSCs中Notch-1、Notch-4和Jagged-1表达的影响。
E-exos在形态、粒径或表面标志物方面与hAMSC-exos无显著差异。E-exos可被HemSCs内化，且E-exos的细胞摄取过程具有时间依赖性。处理12小时后，E-exos显著抑制管腔形成。E-exos通过加载miR-187-3p也抑制了Notch信号通路。E-exos通过Notch信号通路对HemSC增殖显示出优异的抑制作用。
本研究为利用hAMSC-exos优化当前临床治疗方案以促进IH治疗并在未来递送治疗药物奠定了基础。
