Alizadeh Hamid, Muftuoğlu Can, Omondi Zeph Nelson, Mert Ufuk, Asadi Milad, Ozbilgin Ahmet, Caner Ayse
Department of Parasitology, Faculty of Medicine, Ege University, Izmir, Turkey.
Department of Basic Oncology, Institute of Health Sciences, Ege University, Izmir, Turkey.
Acta Trop. 2025 Jan;261:107509. doi: 10.1016/j.actatropica.2024.107509. Epub 2024 Dec 18.
Leishmaniasis is a neglected infectious disease that affects millions of people worldwide. Visceral leishmaniasis (VL) caused by Leishmania infantum and cutaneous leishmaniasis (CL) caused by L. major/ L. tropica are the main clinical forms of this disease, which are life-threatening if not diagnosed and treated properly. Considering the problems in sampling and laboratory diagnosis of leishmaniasis, new molecular markers such as circular RNAs (circRNAs) are needed. circRNAs, a novel class of RNAs, have been one of the most promising targets for the diagnosis and prognosis of diseases. Although the therapeutic and diagnostic role of circRNAs in many diseases and some parasitic diseases are known, not much research has been done in the field of leishmaniasis. We determined the gene expressions of circRNAs in human leukemia monocytic (THP-1) cells after infection with Leishmania. For this, the human cell line THP-1 was differentiated into macrophages by Phorbol 12-myristate 13-acetate (PMA) treatment. Differentiated THP-1 cells were infected with L. infantum and L. tropica promastigotes. After 24 hours, expression levels of circRNAs were determined by RT-qPCR technique. Also, the microRNAs associated with differentially expressed circRNAs were investigated. Then, the molecular pathways associated with expressed circRNAs were obtained by GO and Reactome. The results showed that five circRNAs were differentially expressed in THP1 macrophages infected with L. infantum and L. tropica. These findings suggest that some circRNAs may be potential biomarkers for diagnosis in Leishmania-infected patients. The enrichment analysis revealed that differentially expressed circRNAs are mainly involved in the regulation of protein stability, RNA catabolic process, and P53/PTK6 signaling mechanism. This is the first study to report an overview of Leishmania-induced circRNAs, which can be potential biomarker candidate for diagnosis especially at species level. Notably, expression of some circRNAs in supernatant of Leishmania infected macrophages suggests that these genes are available in body fluids, therefore, can easily be accessed from the patient without invasive methods especially during treatment monitoring.
利什曼病是一种被忽视的传染病，影响着全球数百万人。由婴儿利什曼原虫引起的内脏利什曼病（VL）和由硕大利什曼原虫/热带利什曼原虫引起的皮肤利什曼病（CL）是该疾病的主要临床形式，如果不进行正确诊断和治疗，会危及生命。考虑到利什曼病在采样和实验室诊断方面存在的问题，需要新的分子标记物，如环状RNA（circRNA）。circRNA是一类新型RNA，一直是疾病诊断和预后最有前景的靶点之一。尽管circRNA在许多疾病和一些寄生虫病中的治疗和诊断作用已为人所知，但在利什曼病领域的研究还不多。我们测定了利什曼原虫感染后人白血病单核细胞（THP-1）细胞中circRNA的基因表达。为此，通过佛波酯12-肉豆蔻酸酯13-乙酸酯（PMA）处理将人细胞系THP-1分化为巨噬细胞。分化后的THP-1细胞用婴儿利什曼原虫和热带利什曼原虫前鞭毛体感染。24小时后，通过RT-qPCR技术测定circRNA的表达水平。此外，还研究了与差异表达的circRNA相关的微小RNA。然后，通过基因本体论（GO）和Reactome获得与表达的circRNA相关的分子途径。结果表明，在感染婴儿利什曼原虫和热带利什曼原虫的THP1巨噬细胞中有5种circRNA差异表达。这些发现表明，一些circRNA可能是利什曼原虫感染患者诊断的潜在生物标志物。富集分析显示，差异表达的circRNA主要参与蛋白质稳定性调节、RNA分解代谢过程以及P53/PTK6信号机制。这是第一项报道利什曼原虫诱导的circRNA概述的研究，这些circRNA可能是诊断的潜在生物标志物候选物，尤其是在物种水平上。值得注意的是，利什曼原虫感染的巨噬细胞上清液中一些circRNA的表达表明这些基因存在于体液中，因此，在治疗监测期间，无需侵入性方法就能很容易地从患者体内获取这些基因。
