Developmental regulation and functional prediction of microRNAs in an expanded Fasciola hepatica miRNome
Developmental regulation and functional prediction of microRNAs in an expanded Fasciola hepatica miRNome
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扩展肝片形吸虫 miRNome 中 microRNA 的发育调控和功能预测
DOI：
10.1101/2021.11.08.467689
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Herron C
中科院分区：
文献类型：
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作者：
Herron C
肝片形吸虫（Fasciola hepatica）是对养殖反刍动物的健康和生产力的全球性负担，并且是对人类健康的人畜共患病威胁。尽管明显需要加速发现这种病原体的新药物和疫苗治疗，但我们对肝吸虫生物学和宿主相互作用的了解仍然相对有限。非编码RNA，包括微小（mi）RNA，是所有真核生物中转录调控的关键，因此对miRNA生物学的理解可以在系统水平上揭示生物体功能。四个先前的出版物报道了多达89个成熟的miRNA序列从F。hepatica，但我们的数据表明，这并不代表这个物种miRNome的完整说明。我们已经扩大了以前的研究测序，首次，miRNA从多个生命阶段（成人，新excysted少年（NEJ），囊蚴和成人衍生的细胞外囊泡（EV））。这些实验检测到另外61个高置信度的miRNAs，其中大多数在任何其他物种中都没有描述过，扩大了F。hepaticamiRNA组至150个成熟序列。我们使用定量（q）PCR检测，以提供第一个发育概况的miRNA表达的囊蚴，NEJ，成人和成人衍生的EV。大多数miRNA在囊蚴中表达最高，在整个生命阶段中至少有六个明显的不同表达簇。对细胞内miRNA进行功能分析，以鉴定在肝纤维化组织转录组中具有负相关表达的靶mRNA，突出显示与关键毒力转录物（包括组织蛋白酶蛋白酶）和控制寄生虫生长、发育和运动的神经肌肉基因的调控相互作用。我们还将28个成人来源的EV miRNA与来自反刍动物淋巴结、外周血单核细胞（PBMC）和肝组织转录组的肝片吸虫感染的转录组中的397个宿主基因的下调相关联。这些包括参与信号转导、免疫和代谢途径的基因，增加了片形吸虫病期间基于miRNA的免疫抑制的证据。这些数据扩展了我们对F. hepaticamiRNome，提供了该物种发育miRNA调控的第一个数据，并为肝吸虫miRNA生物学的功能基因组学研究提供了一套可检验的假设。
The liver fluke,Fasciola hepatica, is a global burden on the wellbeing and productivity of farmed ruminants, and a zoonotic threat to human health. Despite the clear need for accelerated discovery of new drug and vaccine treatments for this pathogen, we still have a relatively limited understanding of liver fluke biology and host interactions. Noncoding RNAs, including micro (mi)RNAs, are key to transcriptional regulation in all eukaryotes, such that an understanding of miRNA biology can shed light on organismal function at a systems level. Four previous publications have reported up to 89 mature miRNA sequences fromF. hepatica, but our data show that this does not represent a full account of this species miRNome. We have expanded on previous studies by sequencing, for the first time, miRNAs from multiple life stages (adult, newly excysted juvenile (NEJ), metacercariae and adult-derived extracellular vesicles (EVs)). These experiments detected an additional 61 high-confidence miRNAs, most of which have not been described in any other species, expanding theF. hepaticamiRNome to 150 mature sequences. We used quantitative (q)PCR assays to provide the first developmental profile of miRNA expression across metacercariae, NEJ, adult and adult-derived Evs. The majority of miRNAs were expressed most highly in metacercariae, with at least six distinct expression clusters apparent across life stages. Intracellular miRNAs were functionally analyzed to identify target mRNAs with inversely correlated expression inF. hepaticatissue transcriptomes, highlighting regulatory interactions with key virulence transcripts including cathepsin proteases, and neuromuscular genes that control parasite growth, development and motility. We also linked 28 adult-derived EV miRNAs with downregulation of 397 host genes inF. hepatica-infected transcriptomes from ruminant lymph node, peripheral blood mononuclear cell (PBMC) and liver tissue transcriptomes. These included genes involved in signal transduction, immune and metabolic pathways, adding to the evidence for miRNA-based immunosuppression during fasciolosis. These data expand our understanding of theF. hepaticamiRNome, provide the first data on developmental miRNA regulation in this species, and provide a set of testable hypotheses for functional genomics interrogations of liver fluke miRNA biology.