DataSheet_2_Characterization of MicroRNA Cargo of Extracellular Vesicles Isolated From the Plasma of Schistosoma japonicum-Infected Mice.xlsx

Schistosoma is a genus of parasitic trematodes that undergoes complex migration in final hosts, finally developing into adult worms, which are responsible for egg production and disease dissemination. Recent studies documented the importance of extracellular vesicles (EVs) in the regulation of host-parasite interactions. Herein, we investigated the microRNA (miRNA) profiles of EVs isolated from host plasma at different stages of Schistosoma japonicum infection (lung stage: 3 days post-infection (dpi), and liver stages: 14 and 21 dpi) to identify miRNA cargo potentially involved in the pathogenesis and immune regulation of schistosomiasis. Characterization of the isolated plasma EVs revealed their diameter to be approximately 100 nm, containing typical EV markers such as Hsp70 and Tsg101. Deep sequencing analysis indicated the presence of 811 known and 15 novel miRNAs with an increasing number of differential miRNAs from the lung stage (27 miRNAs) to the liver stages (58 and 96 miRNAs at 14 and 21 dpi, respectively) in the plasma EVs of infected mice compared to EVs isolated from the uninfected control. In total, 324 plasma EV miRNAs were shown to be co-detected among different stages of infection and the validation of selected miRNAs showed trends of abundance similar to deep sequencing analysis. For example, miR-1a-3p and miR-122-5p showed higher abundance, whereas miR-150-3p and miR-126a showed lower abundance in the plasma EVs of infected mice at 3, 14, and 21 dpi as compared to those of uninfected mice. In addition, bioinformatic analysis combined with PCR validation of the miRNA targets, particularly those associated with the immune system and parasitic infectious disease, indicated a significant increase in the expression of Gbp7and Ccr5 in contrast to the decreased expression of Fermt3, Akt1, and IL-12a. Our results suggested that the abundance of miRNA cargo of the host plasma EVs was related to the stages of Schistosoma japonicum infection. Further studies on the roles of these miRNAs may reveal the regulatory mechanism of the host-parasite interaction. Moreover, the differentially abundant miRNA cargo in host EVs associated with S. japonicum infection may also provide valuable clues for identifying novel biomarkers for schistosomiasis diagnosis.

血吸虫属（Schistosoma）是一类寄生性吸虫，其在终宿主体内会经历复杂的迁移过程，最终发育为成虫并介导产卵与疾病传播。近期研究证实，细胞外囊泡（extracellular vesicles, EVs）在调控宿主-寄生虫相互作用中发挥关键作用。本研究针对日本血吸虫（Schistosoma japonicum）感染不同阶段的宿主血浆分离的细胞外囊泡的微小RNA（microRNA, miRNA）谱展开分析，以期筛选可能参与血吸虫病发病与免疫调控的miRNA携带组分。研究中，我们分别检测了感染后3天（肺期，days post-infection, dpi）、14天及21天（肝期）的感染小鼠血浆EVs的miRNA谱，并与未感染对照组的EVs进行对比。对分离得到的血浆EVs进行表征后发现，其粒径约为100 nm，且表达热休克蛋白70（Hsp70）、肿瘤易感基因101（Tsg101）等典型EVs标志物。深度测序分析显示，相较于未感染对照组，感染小鼠血浆EVs中可检测到811种已知miRNA与15种新型miRNA；从肺期（27种差异miRNA）到肝期（感染后14天时有58种，感染后21天时有96种），差异表达miRNA的数量逐渐升高。不同感染阶段中共检出324种血浆EVs源性miRNA，对部分筛选得到的miRNA进行验证，其丰度变化趋势与深度测序结果一致。例如，与未感染小鼠相比，感染后3、14、21 dpi的小鼠血浆EVs中，miR-1a-3p与miR-122-5p的丰度更高，而miR-150-3p与miR-126a的丰度更低。此外，结合生物信息学分析与针对miRNA靶基因的聚合酶链式反应（PCR）验证（尤其是与免疫系统及寄生性传染病相关的靶基因）发现，Gbp7与Ccr5的表达水平显著上调，而Fermt3、Akt1及IL-12a的表达水平显著下调。本研究结果表明，宿主血浆EVs所携带的miRNA组分丰度与日本血吸虫感染阶段密切相关。对这些miRNA的功能开展进一步研究，有望阐明宿主-寄生虫相互作用的调控机制。此外，与日本血吸虫感染相关的宿主EVs差异丰度miRNA组分，也可为开发新型血吸虫病诊断生物标志物提供重要线索。