Anuntasomboon Pornchai, Siripattanapipong Suradej, Unajak Sasimanas, Choowongkomon Kiattawee, Burchmore Richard, Leelayoova Saovanee, Mungthin Mathirut, E-Kobon Teerasak
Department of Genetics, Faculty of Science, Kasetsart University, Bangkok, Thailand.
Omics Center for Agriculture, Bioresources, Food, and Health, Kasetsart University (OmiKU), Bangkok, Thailand.
PLoS Negl Trop Dis. 2024 Dec 17;18(12):e0012716. doi: 10.1371/journal.pntd.0012716. eCollection 2024 Dec.
Amphotericin B (AmB) is a potent antifungal and antiparasitic medication that exerts its action by disrupting the cell membrane of the leishmanial parasite, leading to its death. Understanding the genetic alterations induced by Amphotericin B is crucial for gaining insights into drug resistance mechanisms and developing more effective treatments against Leishmania infections. As a new Leishmania species, the molecular response of Leishmania orientalis to anti-leishmanial drugs has not been fully explored. In this study, Leishmania orientalis strain PCM2 culture was subjected to AmB exposure at a concentration of 0.03 uM over 72 hours compared to the control. The genomic alteration and transcriptomic changes were investigated by utilising the whole genome and RNA sequencing methods, followed by the analysis of single nucleotide polymorphisms (SNPs), differential gene expression, and chromosomal copy number variations (CNVs) assessed using read depth coverage (RDC) values across the entire genome. The chromosomal CNV analysis showed no significant difference between L. orientalis from the control and AmB-treated groups. The distribution of SNPs displayed notable variability, with higher SNP incidence in the control group compared to the AmB-treated group. Gene ontology analysis unveiled functions of the SNPs -associated genes involved in transporter function, genetic precursor synthesis, and purine nucleotide metabolism. Notably, the impact of AmB treatment on the L. orientalis gene expression profiles exhibited diverse expressional alterations, particularly the downregulation of pivotal genes such as the tubulin alpha chain gene. The intricate interplay between SNPs and gene expression alterations might underscore the complex regulatory networks underlying the AmB resistance of L. orientalis strain PCM2.
两性霉素B（AmB）是一种强效的抗真菌和抗寄生虫药物，它通过破坏利什曼原虫的细胞膜发挥作用，导致其死亡。了解两性霉素B诱导的基因改变对于深入了解耐药机制以及开发针对利什曼原虫感染的更有效治疗方法至关重要。作为一种新的利什曼原虫物种，东方利什曼原虫对抗利什曼原虫药物的分子反应尚未得到充分研究。在本研究中，将东方利什曼原虫PCM2菌株培养物与对照组相比，在72小时内暴露于浓度为0.03 μM的两性霉素B中。通过使用全基因组和RNA测序方法研究基因组改变和转录组变化，随后分析单核苷酸多态性（SNP）、差异基因表达以及使用全基因组的读深度覆盖（RDC）值评估的染色体拷贝数变异（CNV）。染色体CNV分析显示对照组和两性霉素B处理组的东方利什曼原虫之间没有显著差异。SNP的分布显示出显著的变异性，对照组中的SNP发生率高于两性霉素B处理组。基因本体分析揭示了与SNP相关的基因在转运蛋白功能、遗传前体合成和嘌呤核苷酸代谢中的功能。值得注意的是，两性霉素B处理对东方利什曼原虫基因表达谱的影响表现出多种表达改变，特别是关键基因如微管蛋白α链基因的下调。SNP与基因表达改变之间的复杂相互作用可能突出了东方利什曼原虫PCM2菌株对两性霉素B耐药性背后的复杂调控网络。
