Department of Public Health and Infectious Diseases, Sapienza University of Rome, Piazzale Aldo Moro 5, 00185, Rome, Italy.
Sci Rep. 2022 Jun 11;12(1):9667. doi: 10.1038/s41598-022-13594-3.
Anisakids are widespread marine parasites of medical, veterinary and economic relevance. They infect marine natural hosts but humans can accidentally acquire the fish-borne zoonosis anisakiasis by ingesting infected raw fishes or mollusks. Among the several species described, Anisakis pegreffii is one of the main etiological agent of the disease, in particular in the Mediterranean area. Despite the growing evidence of miRNAs involvement in host-parasite interplay, and the emerging role of exosomal microvesicles in shuttling them between different cell types (and sometime across species), no information on miRNAs from any Anisakis species is presently available. In this study we isolated extracellular vesicles (EVs) released by Anisakis pegreffii infective third-stage larvae (L3) and analyzed by RNA-seq small RNAs from both L3 and EVs. We showed by nanoparticle tracking analysis that L3 release in culture medium particles of size compatible with the one of extracellular vesicles. A catalogue of 156 miRNAs from A. pegreffii was compiled by sequence comparison to evolutionary close species and miRNA prediction software. Using differential expression analysis, we identified a small number of highly abundant miRNAs in larvae and extracellular vesicles fractions whose potential biological relevance may deserve future investigation. Finally, A. pegreffii miRNAs were compared to those described in other parasitic helminths and predicted targets among human genes were searched, suggesting their potential involvement during infection.
蛔目寄生虫广泛存在于海洋中，具有医学、兽医和经济重要性。它们感染海洋自然宿主，但人类通过摄入受感染的生鱼或贝类，可能会意外获得鱼源性人畜共患寄生虫病——异尖线虫病。在已描述的几种物种中，海狮蛔线虫是该疾病的主要病原体之一，特别是在地中海地区。尽管越来越多的证据表明 microRNAs 参与宿主-寄生虫相互作用，并且外泌体微泡在将它们在不同细胞类型（有时甚至在物种之间）中穿梭的作用不断涌现，但目前尚无任何关于异尖线虫物种 microRNAs 的信息。在这项研究中，我们分离了感染性第三期幼虫（L3）释放的海狮蛔线虫细胞外囊泡（EVs），并通过 RNA-seq 对 L3 和 EVs 中的小 RNA 进行了分析。我们通过纳米颗粒跟踪分析表明，L3 在培养物中释放的颗粒大小与细胞外囊泡的大小相匹配。通过与进化上相近的物种进行序列比较和 microRNA 预测软件，我们编制了一份包含 156 个海狮蛔线虫 microRNAs 的目录。通过差异表达分析，我们在幼虫和细胞外囊泡部分鉴定出少数高度丰富的 microRNAs，它们的潜在生物学意义可能值得进一步研究。最后，我们将海狮蛔线虫的 microRNAs 与其他寄生虫蠕虫中描述的 microRNAs 进行了比较，并搜索了人类基因中的预测靶标，表明它们可能在感染过程中发挥作用。
