Identification and classification of hubs in miRNA target gene networks in human neural stem/progenitor cells following Japanese encephalitis virus infection

Abstract

Micro RNA dysregulation is observed in many viral diseases. RNA viruses modulate host miRNA machinery for their own benefit. JEV, a neurotropic RNA virus has been reported to manipulate several miRNAs in neuron or microglia. However, no report indicates a complete sketch of the miRNA profile of NSPCs contributing to viral persistence; hence being focused in our current study. We performed a miRNA array of 84 miRNAs in human neuronal progenitor cell line and primary neural precursor cells isolated from aborted foetus. Several fold down-regulation of hsa-miR-9-5p, hsa-miR-22-3p, hsa-miR-124-3p and hsa-miR-132-3p were found in both of the cells. Subsequently, we screened for the target genes of these miRNAs and looked for the important biological pathways the genes significantly regulate. Then we sorted out the target genes which are involved in two or more than two pathways. We constructed a protein-protein interaction (PPI) network of the miRNA target genes based on their interaction patterns. A binary adjacency matrix for each gene network was prepared. Different modules or communities were identified in those networks using community detection algorithms. Mathematically, we identified the hub genes by analyzing their degree centrality and participation co-efficient in the network. The hub genes were classified either as provincial (P<0.4) or connector hubs (P>0.4). We validated the expression of hub genes in both cell line and primary cells through qRT-PCR post JEV infection and respective miR-mimic transfection. Taken together, our findings highlight the importance of specific target gene networks of miRNAs affected by JEV infection in NSPCs.\n\nImportanceJEV damages the neural stem/progenitor cell population of mammalian brain. However, JEV induced alteration in miRNA expression pattern of the cell population remains an open question, hence warrants our present study. In this study, we specifically address the down-regulation of four miRNAs and we prepared a protein-protein interaction network of miRNA target genes. We identified two types of hub genes in the PPI network namely connector hubs and provincial hubs. These two types of miRNA target hub genes critically influence the participation strength in the networks and thereby significantly influence up and down regulation in several key biological pathways. Computational analysis of the PPI networks identifies key protein interactions and hubs in those modules which opens up the possibility of precise identification and classification of host factors for viral infection in NSPCs and how RNA viruses modulate host miRNA machinery for their own benefit post JEV infection to the cells.