Akand Sajjadul Kadir, Rahman Areeba, Masood Mohammad, Tabrez Shams, Saleem Mohammad, Ahmed Mohammad Z, Akhter Yusuf, Haque Mohammad Mahfuzul, Rub Abdur
Department of Biotechnology, Jamia Millia Islamia (A Central University), New Delhi, 110025, India.
Department of Pathology, University of Michigan School of Medicine, Ann Arbor, MI, 48109, USA.
Arch Microbiol. 2025 Apr 16;207(6):123. doi: 10.1007/s00203-025-04325-z.
Leishmaniasis, caused by the protozoan parasites of the genus Leishmania, poses a significant global health challenge, particularly in the resource-limited regions where it causes high mortality. Regardless in the progress of treatment strategies, the emergence of drug resistance and limited efficacy requires the search of novel therapy and therapeutic targets. MicroRNAs, the crucial post-transcriptional regulators of gene expression, play critical roles in host-pathogen interactions. Here, we screened the miRNAs dysregulated during Leishmania donovani infection through literature search. hsa-miR-330-5p, one of the miRNAs which through human KEGG 2021 and Human Cyc 2016 analysis was found to be involved in multiple pathways including sphingolipid signaling pathway. Sphingolipids are important class of lipids involved in different cellular processes and therefore are the targets of many pathogens including Leishmania. hsa-miR-330-5p was found downregulated after 24 h of Leishmania donovani infection in THP-1 derived human macrophages. Target prediction of sphingolipid biosynthetic genes through in silico prediction tools showed 3 UTR of serine palmitoyltransferase long chain base subunit 1 to be a target of hsa-miR-330-5p. The in silico target prediction of hsa-miR-330-5p was validated by cloning the 3 UTR target sequence of gene, transfecting and performing luciferase assay in HEK 293 T cell line. Transfection of mimic of hsa-miR-330-5p reduced the luciferase activity which validated the in silico target prediction. Further, mimic of hsa-miR-330-5p inhibited the expression of the target gene, serine palmitoyltransferase long chain base subunit 1 and augmented the expression of pro-inflammatory cytokines in L. donovani infected THP-1 derived macrophages. Mimic of hsa-miR-330-5p also led to a significant reduction in the intracellular parasite burden in both THP-1 derived as well as primary human macrophages. This study has not only identified the sphingolipid biosynthesis regulatory miRNA but will also help in the development of novel and effective treatment strategy against leishmaniasis in future.
利什曼病由利什曼原虫属的原生动物寄生虫引起，是一项重大的全球健康挑战，在资源有限地区尤其如此，因为该病在这些地区会导致高死亡率。尽管治疗策略有所进展，但耐药性的出现和疗效有限仍需要寻找新的治疗方法和治疗靶点。微小RNA是基因表达的关键转录后调节因子，在宿主与病原体的相互作用中发挥着关键作用。在这里，我们通过文献检索筛选了杜氏利什曼原虫感染过程中失调的微小RNA。hsa-miR-330-5p是通过人类KEGG 2021和人类Cyc 2016分析发现参与包括鞘脂信号通路在内的多种途径的微小RNA之一。鞘脂是参与不同细胞过程的重要脂质类别，因此是包括利什曼原虫在内的许多病原体的靶点。在THP-1衍生的人类巨噬细胞中，杜氏利什曼原虫感染24小时后，hsa-miR-330-5p被发现下调。通过计算机预测工具对鞘脂生物合成基因进行靶标预测，结果显示丝氨酸棕榈酰转移酶长链碱基亚基1的3'UTR是hsa-miR-330-5p的靶标。通过克隆该基因的3'UTR靶标序列、转染并在HEK 293T细胞系中进行荧光素酶测定，验证了hsa-miR-330-5p的计算机靶标预测。转染hsa-miR-330-5p模拟物降低了荧光素酶活性，这验证了计算机靶标预测。此外，hsa-miR-330-5p模拟物抑制了靶基因丝氨酸棕榈酰转移酶长链碱基亚基1的表达，并增强了杜氏利什曼原虫感染的THP-1衍生巨噬细胞中促炎细胞因子的表达。hsa-miR-330-5p模拟物还导致THP-1衍生的巨噬细胞以及原代人类巨噬细胞中的细胞内寄生虫负荷显著降低。这项研究不仅鉴定了鞘脂生物合成调节性微小RNA，还将有助于未来开发针对利什曼病的新型有效治疗策略。
