Research Center for Hydatid Disease in Iran, Kerman University of Medical Sciences, Kerman, Iran.
Department of Agricultural Biotechnology, Faculty of Agriculture, Shahid Bahonar University of Kerman, Kerman, Iran.
Parasitol Res. 2021 Sep;120(9):3203-3214. doi: 10.1007/s00436-021-07251-3. Epub 2021 Aug 5.
MicroRNAs are critical gene regulators at the post-transcriptional level and play essential roles in numerous developmental processes in metazoan parasites including the causative agent of cystic echinococcosis, Echinococcus granulosus. The molecular basis of different patterns of E. granulosus development in the canine definitive host and in in vitro culture systems is poorly understood. In the present study, miRNA transcriptomes of the strobilated worms derived from experimental infection in the definitive host were compared with those from diphasic culture system after 60-day protoscoleces cultivation. Total RNA was extracted from in vivo- and in vitro-derived strobilated worms. Small RNA libraries were constructed, and deep sequencing was performed. Subsequently, differential miRNA expressions and target predictions were obtained, and pathway analysis was performed by gene ontology and KEGG. Seven miRNAs were differentially expressed between the in vivo- and in vitro-derived worms. In addition, we reported 13 novel miRNA candidates and 42 conserved miRNAs. Four out of five top miRNAs with the highest read counts were shared between the in vivo and in vitro-derived worms, i.e., egr-miR-10a-5p, egr-let-7-5p, egr-bantam-3p, and egr-miR-71-5p. Target prediction of the differential miRNAs between the two systems showed significant differences in the membrane-enclosed lumen, membrane part, and an intrinsic component of the membrane. Findings of KEGG analysis indicated that differentially expressed miRNAs were involved in hippo, MAPK, and WNT signaling pathways. The study demonstrated a significant difference in miRNA transcriptomes and related signaling pathways between the two systems, suggesting the importance of host-parasite interplay in the fate of protoscoleces development in in vivo and in vitro systems.
微小 RNA 是转录后水平的关键基因调控因子,在后生动物寄生虫的许多发育过程中发挥着重要作用,包括包虫病的病原体棘球蚴。犬科终宿主和体外培养系统中棘球蚴发育不同模式的分子基础知之甚少。在本研究中,比较了来自终宿主实验感染的具纤毛幼虫与 60 天原头蚴培养后的双相培养系统的 miRNA 转录组。从体内和体外来源的具纤毛幼虫中提取总 RNA。构建了小 RNA 文库,并进行了深度测序。随后,获得了差异 miRNA 表达和靶标预测,并通过基因本体和 KEGG 进行了通路分析。在体内和体外来源的蠕虫之间有 7 个 miRNA 表达差异。此外,我们还报道了 13 个新的 miRNA 候选物和 42 个保守的 miRNA。在体内和体外来源的蠕虫中,5 个最高读数 miRNA 中有 4 个是相同的,即 egr-miR-10a-5p、egr-let-7-5p、egr-bantam-3p 和 egr-miR-71-5p。两个系统之间差异 miRNA 的靶标预测显示,在膜封闭腔、膜部分和膜的固有成分之间存在显著差异。KEGG 分析的结果表明,差异表达的 miRNA 参与了 hippo、MAPK 和 WNT 信号通路。该研究表明,两个系统之间的 miRNA 转录组和相关信号通路存在显著差异,提示宿主-寄生虫相互作用在体内和体外系统中原头蚴发育命运中的重要性。