Transcriptomic analyses implicate neuronal plasticity and chloride homeostasis in ivermectin resistance and response to treatment in a parasitic nematode.
Transcriptomic analyses implicate neuronal plasticity and chloride homeostasis in ivermectin resistance and response to treatment in a parasitic nematode.
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DOI：
10.1371/journal.ppat.1010545
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发表时间：
2022-06
期刊：
影响因子：
6.7
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中科院分区：
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--
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抗寄生虫药物伊维菌素在全球人类和动物健康中发挥着重要作用。然而，伊维菌素耐药性在兽医蠕虫中广泛存在，并且人们越来越关注人类相关蠕虫对治疗的次优反应。尽管几十年的研究，伊维菌素耐药的遗传机制是知之甚少寄生蠕虫。这反映了伊维菌素在寄生蠕虫中的作用方式以及这些生物的遗传复杂性的显著不确定性;寄生蠕虫具有大的、快速进化的基因组，进化历史和遗传背景的差异可能混淆抗性和易感群体之间的比较。我们进行了一个多药耐药和敏感的参考分离的捻转血矛线虫，一个经济上重要的胃肠道线虫的羊，伊维菌素选择的F2人口与未处理的F2对照进行比较的控制遗传杂交。对所有种群的雄性和雌性成虫进行的RNA-seq分析确定了亲本分离株之间的高转录组分化，该分化在F2中显著降低，从而确定了与伊维菌素抗性特异性相关的差异。在所有的耐药人群中，有一个单一的基因，HCON_00155390：cky-1，一个假定的咽表达的转录因子，在一个狭窄的基因座上的染色体V先前被证明是伊维菌素的选择组成性上调。此外，我们检测到耐药和易感人群之间基因表达的性别特异性差异，包括仅在耐药男性中P-糖蛋白HCON_00162780：pgp-11的组成性上调。伊维菌素选择后，我们确定了差异表达的基因在神经元功能和氯稳态，这是一致的适应性反应伊维菌素诱导的神经肌肉细胞超极化的作用。总的来说，我们展示了遗传杂交的效用，以确定伊维菌素选择特异性的基因表达差异，并提供了一个框架，以更好地了解伊维菌素抗性和寄生蠕虫对治疗的反应。寄生蠕虫（蠕虫）感染世界各地的人和动物，并在很大程度上通过大规模施用驱虫药物得到控制。可用的驱虫药数量非常有限，寄生蠕虫可迅速对这些药物产生耐药性。伊维菌素是一种广泛用于人类和动物的驱虫药，但目前在兽医领域普遍存在耐药性。我们将伊维菌素耐药和伊维菌素敏感的寄生蠕虫杂交，用伊维菌素处理它们或将它们作为未处理的对照。这为研究伊维菌素抗性相关基因表达差异提供了相似遗传背景的抗性和易感人群。我们在所有耐药种群的雄性和雌性蠕虫中发现了一个先前与耐药性无关的基因（HCON_00155390：cky-1）上调。这种基因被认为在蠕虫的咽部（口器）中表达，在哺乳动物中，在控制神经功能和保护神经免受损伤方面发挥作用。这与伊维菌素通过咽麻痹抑制蠕虫进食的已知作用一致，可能涉及一种新的机制，使耐药蠕虫在治疗中存活。
The antiparasitic drug ivermectin plays an essential role in human and animal health globally. However, ivermectin resistance is widespread in veterinary helminths and there are growing concerns of sub-optimal responses to treatment in related helminths of humans. Despite decades of research, the genetic mechanisms underlying ivermectin resistance are poorly understood in parasitic helminths. This reflects significant uncertainty regarding the mode of action of ivermectin in parasitic helminths, and the genetic complexity of these organisms; parasitic helminths have large, rapidly evolving genomes and differences in evolutionary history and genetic background can confound comparisons between resistant and susceptible populations. We undertook a controlled genetic cross of a multi-drug resistant and a susceptible reference isolate of Haemonchus contortus, an economically important gastrointestinal nematode of sheep, and ivermectin-selected the F2 population for comparison with an untreated F2 control. RNA-seq analyses of male and female adults of all populations identified high transcriptomic differentiation between parental isolates, which was significantly reduced in the F2, allowing differences associated specifically with ivermectin resistance to be identified. In all resistant populations, there was constitutive upregulation of a single gene, HCON_00155390:cky-1, a putative pharyngeal-expressed transcription factor, in a narrow locus on chromosome V previously shown to be under ivermectin selection. In addition, we detected sex-specific differences in gene expression between resistant and susceptible populations, including constitutive upregulation of a P-glycoprotein, HCON_00162780:pgp-11, in resistant males only. After ivermectin selection, we identified differential expression of genes with roles in neuronal function and chloride homeostasis, which is consistent with an adaptive response to ivermectin-induced hyperpolarisation of neuromuscular cells. Overall, we show the utility of a genetic cross to identify differences in gene expression that are specific to ivermectin selection and provide a framework to better understand ivermectin resistance and response to treatment in parasitic helminths. Parasitic helminths (worms) infect people and animals throughout the world and are largely controlled with mass administration of anthelmintic drugs. There are a very limited number of anthelmintics available and parasitic helminths can rapidly develop resistance to these drugs. Ivermectin is a widely used anthelmintic in both humans and animals, but resistance is now widespread in the veterinary field. We crossed ivermectin resistant and ivermectin susceptible parasitic helminths and treated them with ivermectin or left them as untreated controls. This provided resistant and susceptible populations with a similar genetic background with which to study differences in gene expression associated with ivermectin resistance. We identified upregulation of a gene with no previous association with drug resistance (HCON_00155390:cky-1) in male and female worms in all resistant populations. This gene is thought to be expressed in the helminth pharynx (mouthpart) and, in mammals, plays a role in controlling nerve function and protecting nerves from damage. This is consistent with the known effects of ivermectin in inhibiting helminth feeding through pharyngeal paralysis and may implicate a novel mechanism that allows resistant worms to survive treatment.
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DOI：
10.1093/bioinformatics/btx346
发表时间：
2017-10-01
期刊：
Bioinformatics (Oxford, England)
影响因子：
--
作者：
Gel B;Serra E
通讯作者：
Serra E
影响因子：
4.5
作者：
Hisamoto N;Nagamori Y;Shimizu T;Pastuhov SI;Matsumoto K
通讯作者：
Matsumoto K
影响因子：
3.3
作者：
Doyle SR;Laing R;Bartley DJ;Britton C;Chaudhry U;Gilleard JS;Holroyd N;Mable BK;Maitland K;Morrison AA;Tait A;Tracey A;Berriman M;Devaney E;Cotton JA;Sargison ND
通讯作者：
Sargison ND
影响因子：
5.3
作者：
Branicky, Robyn;Miyazaki, Hiroaki;Schafer, William R.
通讯作者：
Schafer, William R.
影响因子：
6.7
作者：
Blanchard A;Guégnard F;Charvet CL;Crisford A;Courtot E;Sauvé C;Harmache A;Duguet T;O'Connor V;Castagnone-Sereno P;Reaves B;Wolstenholme AJ;Beech RN;Holden-Dye L;Neveu C
通讯作者：
Neveu C