Goswami Saptamita, Banerjee Atoshi, Kumari Bharti, Bandopadhyay Bhaswati, Bhattacharya Nemai, Basu Nandita, Vrati Sudhanshu, Banerjee Arup
Kolkata School of Tropical Medicine, 108, Chittaranjan Avenue, Kolkata, 700 073, India.
Vaccine and Infectious Disease Research Center (VIDRC), Translational Health Science and Technology Institute (THSTI), NCR Biotech Science Cluster, 3rd Milestone, Faridabad-Gurgaon Expressway, PO box #04, Faridabad, 121001, India.
Mol Neurobiol. 2017 Mar;54(2):1541-1551. doi: 10.1007/s12035-016-9764-y. Epub 2016 Feb 9.
Changes in circulating microRNAs (miRNAs) in the cerebrospinal fluid (CSF) have been associated with different neurological diseases. Here, we presented results of a pilot study aimed at determining the feasibility of detecting miRNAs in the CSF of Japanese Encephalitis virus (JEV) infected individuals with acute encephalitis syndrome (AES). We demonstrated the circulating miRNA profile in CSF of acute encephalitis patients infected with JEV. Using a quantitative real-time PCR-based miRNA array, we examined the level of 87 miRNAs expressed in human exosomes isolated from CSF. Subsequently, correlation between cytokine level and miRNAs expression in CSF samples was examined. In this study, we identified and validated the upregulated expression of three miRNAs, miR-21-5p, miR-150-5p, and miR-342-3p that were specifically circulated in CSF of acute encephalitis patients infected with JEV. CSF miR-21-5p, miR-150-5p, and miR-342-3p expressions were also elevated in infected mice brain. However, the expression pattern of these miRNAs differed in neuronal cells, microglial cells, and the exosome derived from JEV-infected cell culture supernatant. Interestingly, neuronal cells infected with vaccine strain (SA-14-14) did not lead to any upregulation of these three miRNAs. Further, miR-150-5p expression was found to be negatively correlated(r = -0.5279, p = 0.016) with TNFα level. Pathway analysis of putative target genes of these miRNAs indicated involvement of TGF-β, NGF, axon guidance, and MAPK signaling pathways in JEV/AES patients. This study for the first time represents the circulating miRNA in CSF of AES patients and identified the upregulated miRNAs in JEV-infected patients and offers the basis for future investigation.
脑脊液（CSF）中循环微小RNA（miRNA）的变化与不同的神经系统疾病有关。在此，我们展示了一项初步研究的结果，该研究旨在确定检测急性脑炎综合征（AES）的日本脑炎病毒（JEV）感染个体脑脊液中miRNA的可行性。我们展示了JEV感染的急性脑炎患者脑脊液中的循环miRNA谱。使用基于定量实时PCR的miRNA阵列，我们检测了从脑脊液中分离的人外泌体中表达的87种miRNA的水平。随后，检测了脑脊液样本中细胞因子水平与miRNA表达之间的相关性。在本研究中，我们鉴定并验证了三种miRNA，即miR-21-5p、miR-150-5p和miR-342-3p的上调表达，它们在JEV感染的急性脑炎患者脑脊液中特异性循环。感染小鼠脑内脑脊液中miR-21-5p、miR-150-5p和miR-342-3p的表达也升高。然而，这些miRNA在神经元细胞、小胶质细胞以及JEV感染细胞培养上清液来源的外泌体中的表达模式有所不同。有趣的是，用疫苗株（SA-14-14）感染的神经元细胞并未导致这三种miRNA的任何上调。此外，发现miR-150-5p表达与TNFα水平呈负相关（r = -0.5279，p = 0.016）。对这些miRNA的假定靶基因进行的通路分析表明，TGF-β、NGF、轴突导向和MAPK信号通路参与了JEV/AES患者的发病过程。本研究首次展示了AES患者脑脊液中的循环miRNA，并鉴定了JEV感染患者中上调的miRNA，为未来的研究提供了基础。
