Dual microRNA Screens Reveal That the Immune-Responsive miR-181 Promotes Henipavirus Entry and Cell-Cell Fusion

PLoS Pathog. 2016 Oct 26;12(10):e1005974. doi: 10.1371/journal.ppat.1005974. eCollection 2016 Oct.

Abstract

Hendra and Nipah viruses (family Paramyxoviridae, genus Henipavirus) are bat-borne viruses that cause fatal disease in humans and a range of other mammalian species. Gaining a deeper understanding of host pathways exploited by henipaviruses for infection may identify targets for new anti-viral therapies. Here we have performed genome-wide high-throughput agonist and antagonist screens at biosafety level 4 to identify host-encoded microRNAs (miRNAs) impacting henipavirus infection in human cells. Members of the miR-181 and miR-17~93 families strongly promoted Hendra virus infection. miR-181 also promoted Nipah virus infection, but did not affect infection by paramyxoviruses from other genera, indicating specificity in the virus-host interaction. Infection promotion was primarily mediated via the ability of miR-181 to significantly enhance henipavirus-induced membrane fusion. Cell signalling receptors of ephrins, namely EphA5 and EphA7, were identified as novel negative regulators of henipavirus fusion. The expression of these receptors, as well as EphB4, were suppressed by miR-181 overexpression, suggesting that simultaneous inhibition of several Ephs by the miRNA contributes to enhanced infection and fusion. Immune-responsive miR-181 levels was also up-regulated in the biofluids of ferrets and horses infected with Hendra virus, suggesting that the host innate immune response may promote henipavirus spread and exacerbate disease severity. This study is the first genome-wide screen of miRNAs influencing infection by a clinically significant mononegavirus and nominates select miRNAs as targets for future anti-viral therapy development.

MeSH terms

  • Animals
  • Ferrets
  • Fluorescent Antibody Technique
  • Genome-Wide Association Study
  • Henipavirus
  • Henipavirus Infections / genetics*
  • High-Throughput Nucleotide Sequencing
  • Horses
  • Humans
  • MicroRNAs / genetics*
  • Real-Time Polymerase Chain Reaction
  • Virus Internalization*

Substances

  • MIRN-181 microRNA, human
  • MicroRNAs

Grant support

This work was supported by The Commonwealth Scientific and Industrial Research Organisation and the Australian National Health and Medical Research Council (grant 1042452 to CRS). The Victorian Centre for Functional Genomics was funded by the Australian Cancer Research Foundation (ACRF), the Victorian Department of Industry, Innovation and Regional Development (DIIRD), the Australian Phenomics Network (APN) and supported by funding from the Australian Government’s Education Investment Fund through the Super Science Initiative, the Australasian Genomics Technologies Association (AGTA), the Brockhoff Foundation and the Peter MacCallum Cancer Centre Foundation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.