Department of Molecular Biosciences, Northwestern University, Evanston, IL, 60208, USA.
Department of Molecular Biosciences, Northwestern University, Evanston, IL, 60208, USA; Driskill Graduate Program in Life Sciences, Northwestern University, Chicago, IL, 60611, USA.
Int J Parasitol Drugs Drug Resist. 2022 Dec;20:89-96. doi: 10.1016/j.ijpddr.2022.10.004. Epub 2022 Oct 29.
Parasitic nematode infections cause an enormous global burden to both humans and livestock. Resistance to the limited arsenal of anthelmintic drugs used to combat these infections is widespread, including benzimidazole (BZ) compounds. Previous studies using the free-living nematode Caenorhabditis elegans to model parasitic nematode resistance have shown that loss-of-function mutations in the beta-tubulin gene ben-1 confer resistance to BZ drugs. However, the mechanism of resistance and the tissue-specific susceptibility are not well known in any nematode species. To identify in which tissue(s) ben-1 function underlies BZ susceptibility, transgenic strains that express ben-1 in different tissues, including hypodermis, muscles, neurons, intestine, and ubiquitous expression were generated. High-throughput fitness assays were performed to measure and compare the quantitative responses to BZ compounds among different transgenic lines. Significant BZ susceptibility was observed in animals expressing ben-1 in neurons, comparable to expression using the ben-1 promoter. This result suggests that ben-1 function in neurons underlies susceptibility to BZ. Subsetting neuronal expression of ben-1 based on the neurotransmitter system further restricted ben-1 function in cholinergic neurons to cause BZ susceptibility. These results better inform our current understanding of the cellular mode of action of BZs and also suggest additional treatments that might potentiate the effects of BZs in neurons.
寄生虫线虫感染给人类和家畜都带来了巨大的全球负担。用于对抗这些感染的有限驱虫药物 arsenal 的耐药性广泛存在，包括苯并咪唑（BZ）化合物。以前使用自由生活的线虫秀丽隐杆线虫来模拟寄生虫线虫耐药性的研究表明，β-微管蛋白基因 ben-1 的功能丧失突变赋予了 BZ 药物的耐药性。然而，在任何线虫物种中，耐药机制和组织特异性易感性都不清楚。为了确定 ben-1 在哪些组织中发挥作用导致 BZ 易感性，生成了在不同组织（包括皮下组织、肌肉、神经元、肠和普遍表达）中表达 ben-1 的转基因株系。进行了高通量适应性测定，以测量和比较不同转基因系对 BZ 化合物的定量反应。在表达神经元中的 ben-1 的动物中观察到明显的 BZ 易感性，与使用 ben-1 启动子的表达相当。这一结果表明，ben-1 在神经元中的功能是 BZ 易感性的基础。基于神经递质系统进一步限制 ben-1 在胆碱能神经元中的功能，以引起 BZ 易感性，从而对神经元中的 ben-1 表达进行子集化。这些结果更好地告知了我们当前对 BZ 细胞作用模式的理解，并提出了可能增强 BZ 在神经元中效果的其他治疗方法。
