Korhonen Pasi K, Wang Tao, Young Neil D, Byrne Joseph J, Campos Tulio L, Chang Bill C H, Taki Aya C, Gasser Robin B
Department of Veterinary Biosciences, Melbourne Veterinary School, Faculty of Science, The University of Melbourne, Parkville, Victoria 3010, Australia.
Comput Struct Biotechnol J. 2024 Jan 17;23:1026-1035. doi: 10.1016/j.csbj.2024.01.008. eCollection 2024 Dec.
Advances in single cell technologies are allowing investigations of a wide range of biological processes and pathways in animals, such as the multicellular model organism - a free-living nematode. However, there has been limited application of such technology to related parasitic nematodes which cause major diseases of humans and animals worldwide. With no vaccines against the vast majority of parasitic nematodes and treatment failures due to drug resistance or inefficacy, new intervention targets are urgently needed, preferably informed by a deep understanding of these nematodes' cellular and molecular biology - which is presently lacking for most worms. Here, we created the first single cell atlas for an early developmental stage of a highly pathogenic, -related parasitic nematode. We obtained and curated RNA sequence (snRNA-seq) data from single nuclei from embryonating eggs of (150,000 droplets), and selected high-quality transcriptomic data for > 14,000 single nuclei for analysis, and identified 19 distinct clusters of cells. Guided by comparative analyses with , we were able to reproducibly assign seven cell clusters to body wall muscle, hypodermis, neuronal, intestinal or seam cells, and identified eight genes that were transcribed in all cell clusters/types, three of which were inferred to be essential in . Two of these genes (i.e. and ), coding for eukaryotic elongation factors (called eEF1A and -eEF1G), were also demonstrated to be transcribed and expressed in all key developmental stages of Together with these findings, sequence- and structure-based comparative analyses indicated the potential of -eEF1A and/or -eEF1G as intervention targets within the protein biosynthesis machinery of . Future work will focus on single cell studies of all key developmental stages and tissues of , and on evaluating the suitability of the two elongation factor proteins as drug targets in and related nematodes, with a view to finding new nematocidal drug candidates.
单细胞技术的进步使得人们能够对动物体内广泛的生物过程和途径进行研究，比如多细胞模式生物——一种自由生活的线虫。然而，这项技术在相关寄生线虫中的应用有限，而这些寄生线虫在全球范围内引发了人类和动物的重大疾病。由于绝大多数寄生线虫没有疫苗，且因耐药性或无效性导致治疗失败，因此迫切需要新的干预靶点，最好是基于对这些线虫细胞和分子生物学的深入理解——而目前大多数线虫缺乏这方面的了解。在此，我们创建了首个针对一种高致病性相关寄生线虫早期发育阶段的单细胞图谱。我们从（150,000个液滴）胚胎卵的单核中获取并整理了RNA序列（snRNA-seq）数据，选择了超过14,000个单核的高质量转录组数据进行分析，并鉴定出19个不同的细胞簇。通过与[具体物种]的比较分析指导，我们能够将七个细胞簇可重复地分配到体壁肌肉、皮下组织、神经元、肠道或缝合线细胞，并鉴定出在所有细胞簇/类型中都转录的八个基因，其中三个基因被推断在[目标线虫物种]中至关重要。这两个基因（即[基因名称1]和[基因名称2]）编码真核延伸因子（称为eEF1A和-eEF1G），还被证明在[目标线虫物种]的所有关键发育阶段都有转录和表达。结合这些发现，基于序列和结构的比较分析表明，-eEF1A和/或-eEF1G在[目标线虫物种]的蛋白质生物合成机制中具有作为干预靶点的潜力。未来的工作将集中于对[目标线虫物种]所有关键发育阶段和组织的单细胞研究，以及评估这两种延伸因子蛋白作为[目标线虫物种]及相关线虫药物靶点的适用性，以期找到新的杀线虫药物候选物。
