DataSheet_7_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移行过程，最终发育为成虫，负责虫卵产生与疾病传播。多项研究已证实，细胞外囊泡（extracellular vesicles, EVs）在宿主-寄生虫互作调控中发挥重要作用。本研究针对日本血吸虫（Schistosoma japonicum）感染的不同阶段（肺期：感染后3天，即3 dpi；肝期：感染后14天和21天，即14 dpi、21 dpi），对宿主血浆中分离得到的细胞外囊泡的微小RNA（microRNA, miRNA）表达谱进行分析，以期筛选出可能参与血吸虫病发病与免疫调控的miRNA载荷。对分离得到的血浆细胞外囊泡进行表征后发现，其粒径约为100 nm，且表达典型的细胞外囊泡标志物Hsp70与Tsg101。高通量测序分析显示，与未感染对照组的血浆细胞外囊泡相比，感染小鼠血浆细胞外囊泡中共检测到811种已知miRNA和15种新型miRNA；且差异表达miRNA的数量从肺期（27种）逐步升高至肝期（14 dpi时58种，21 dpi时96种）。共计324种血浆细胞外囊泡miRNA在各感染阶段均被共同检测到；对筛选出的miRNA进行验证的结果显示，其表达丰度变化趋势与高通量测序分析结果一致。例如，在感染后3、14、21 dpi的小鼠血浆细胞外囊泡中，miR-1a-3p与miR-122-5p的表达丰度均高于未感染对照组，而miR-150-3p与miR-126a的表达丰度则低于对照组。此外，结合生物信息学分析与PCR验证的miRNA靶基因（尤其是与免疫系统及寄生性传染病相关的靶基因）结果显示，Gbp7与Ccr5的表达水平显著上调，而Fermt3、Akt1及IL-12a的表达水平则显著下调。本研究结果表明，宿主血浆细胞外囊泡的miRNA载荷丰度与日本血吸虫感染阶段密切相关。针对这些miRNA功能的进一步研究，有望阐明宿主-寄生虫互作的调控机制。此外，与日本血吸虫感染相关的宿主细胞外囊泡差异表达miRNA载荷，也可为筛选血吸虫病诊断的新型生物标志物提供重要线索。