背景:扭曲的Haemonchuscontortus(H.contortus)是反刍动物中最常见的寄生线虫,在世界范围内普遍存在。H.扭曲对阿苯达唑(ABZ)的耐药性阻碍了驱虫药的疗效,但是对调节抗药性的分子机制知之甚少。最近的研究表明,长链非编码RNA(lncRNA)可以作为耐药性出现的关键调节因子发挥重要作用。
结果:在这项研究中,对阿苯达唑敏感的(ABZ敏感的)和阿苯达唑耐药的(ABZ耐药的)H.contortus菌株进行了转录组测序,每组有三个生物重复。转录组数据中的lncRNA分析显示,根据|log2Foldchange|≥1和FDR<0.05的标准,ABZ敏感和ABZ抗性菌株之间有276个差异表达的lncRNA(DElncRNA)。值得注意的是,MSTRG.12969.2和MSTRG.9827.1表现出最显着的上调和下调,分别,在抗性菌株中。DElncRNAs的潜在作用包括催化活性,刺激反应,调节药物代谢,和调节免疫反应。此外,我们研究了DElncRNAs和其他RNAs之间的相互作用,特别是MSTRG.12741.1、MSTRG.11848.1、MSTRG.5895.1和MSTRG.14071.1,参与通过顺式/反式/反义/lncRNA-miRNA-mRNA相互作用网络调节药物刺激。这种调节导致相关基因的表达减少(或增加),从而增强了扭曲H.对阿苯达唑的抗性。此外,通过对参与耐药相关途径的竞争性内源性RNA(ceRNA)的综合分析,如mTOR信号通路和ABC转运体信号通路,MSTRG.2499.1-new-m0062-3p-HCON_00099610相互作用的相关性被确定为主要涉及催化活性的调节,新陈代谢,基因启动子的泛素化和转录调控。此外,定量实时聚合酶链反应(qRT-PCR)验证表明,六个DElncRNAs和六个DEmRNAs的转录谱与通过RNA-seq获得的转录谱一致。
结论:本研究的结果使我们能够更好地了解ABZ抗性H.contortus的lncRNA表达谱的变化。总的来说,这些结果表明,lncRNAsMSTRG.963.1,MSTRG.12741.1,MSTRG.11848.1和MSTRG.2499.1在ABZ抗性的发展中起重要作用,并且可以作为有希望的生物标志物用于进一步研究。
BACKGROUND: Haemonchus contortus (H. contortus) is the most common parasitic nematode in ruminants and is prevalent worldwide. H. contortus resistance to albendazole (ABZ) hinders the efficacy of anthelmintic drugs, but little is known about the molecular mechanisms that regulate this of drug resistance. Recent research has demonstrated that long noncoding RNAs (lncRNAs) can exert significant influence as pivotal regulators of the emergence of drug resistance.
RESULTS: In this study, transcriptome sequencing was conducted on both albendazole-sensitive (ABZ-sensitive) and albendazole-resistant (ABZ-resistant) H. contortus strains, with three biological replicates for each group. The analysis of lncRNA in the transcriptomic data revealed that there were 276 differentially expressed lncRNA (DElncRNA) between strains with ABZ-sensitive and ABZ-resistant according to the criteria of |log2Foldchange|≥ 1 and FDR < 0.05. Notably, MSTRG.12969.2 and MSTRG.9827.1 exhibited the most significant upregulation and downregulation, respectively, in the resistant strains. The potential roles of the DElncRNAs included catalytic activity, stimulus response, regulation of drug metabolism, and modulation of the immune response. Moreover, we investigated the interactions between DElncRNAs and other RNAs, specifically MSTRG.12741.1, MSTRG.11848.1, MSTRG.5895.1, and MSTRG.14070.1, involved in regulating drug stimulation through cis/trans/antisense/lncRNA‒miRNA-mRNA interaction networks. This regulation leads to a decrease (or increase) in the expression of relevant genes, consequently enhancing the resistance of H. contortus to albendazole. Furthermore, through comprehensive analysis of competitive endogenous RNAs (ceRNAs) involved in drug resistance-related pathways, such as the mTOR signalling pathway and ABC transporter signalling pathway, the relevance of the MSTRG.2499.1-novel-m0062-3p-HCON_00099610 interaction was identified to mainly involve the regulation of catalytic activity, metabolism, ubiquitination and transcriptional regulation of gene promoters. Additionally, quantitative real-time polymerase chain reaction (qRT-PCR) validation indicated that the transcription profiles of six DElncRNAs and six DEmRNAs were consistent with those obtained by RNA-seq.
CONCLUSIONS: The results of the present study allowed us to better understand the changes in the lncRNA expression profile of ABZ-resistant H. contortus. In total, these results suggest that the lncRNAs MSTRG.963.1, MSTRG.12741.1, MSTRG.11848.1 and MSTRG.2499.1 play important roles in the development of ABZ resistance and can serve as promising biomarkers for further study.