Genetic depletion studies inform receptor usage by virulent hantaviruses in human endothelial cells.

Maria Eugenia Dieterle,Ezgi Kasikci,Chunyan Ye,Carles Solà-Riera,Jonas Klingström,Rohit K Jangra,Steven B Bradfute,Eva Mittler,Kartik Chandran,Samuel M Goodfellow

doi:10.7554/elife.69708

Abstract

Hantaviruses are RNA viruses with known epidemic threat and potential for emergence. Several rodent-borne hantaviruses cause zoonoses accompanied by severe illness and death. However, assessments of zoonotic risk and the development of countermeasures are challenged by our limited knowledge of the molecular mechanisms of hantavirus infection, including the identities of cell entry receptors and their roles in influencing viral host range and virulence. Despite the long-standing presumption that β3/β1-containing integrins are the major hantavirus entry receptors, rigorous genetic loss-of-function evidence supporting their requirement, and that of decay-accelerating factor (DAF), is lacking. Here, we used CRISPR/Cas9 engineering to knockout candidate hantavirus receptors, singly and in combination, in a human endothelial cell line that recapitulates the properties of primary microvascular endothelial cells, the major targets of viral infection in humans. The loss of β3 integrin, β1 integrin, and/or DAF had little or no effect on entry by a large panel of hantaviruses. By contrast, loss of protocadherin-1, a recently identified entry receptor for some hantaviruses, substantially reduced hantavirus entry and infection. We conclude that major host molecules necessary for endothelial cell entry by PCDH1-independent hantaviruses remain to be discovered.

Highlights

Hantaviruses are a large family of enveloped viruses with segmented negative-strand RNA genomes whose members infect a wide range of mammalian hosts (Elliott et al, 1991; Mittler et al, 2019; Vaheri et al, 2013)
Most hemorrhagic fever with renal syndrome (HFRS) cases are associated with the ‘Old-World hantaviruses’ Hantaan virus (HTNV), SEOV, Dobrava-Belgrade virus (DOBV), and Puumala virus (PUUV) infections, whereas hantavirus cardiopulmonary syndrome (HCPS) is primarily caused by the ‘New-World hantaviruses’ Andes virus (ANDV) and Sin Nombre virus (SNV) (Jiang et al, 2016; Macneil et al, 2011)
We used a genetic depletion/complementation strategy to investigate the requirements for four hantavirus receptor candidates/entry host factors described in the literature – b3 integrin, b1 integrin, decay-accelerating factor (DAF), and PCDH1 – during glycoprotein-mediated entry and infection of human endothelial cells by divergent hantaviruses

Summary

Introduction

Hantaviruses are a large family of enveloped viruses with segmented negative-strand RNA genomes whose members infect a wide range of mammalian hosts (Elliott et al, 1991; Mittler et al, 2019; Vaheri et al, 2013). We identified protocadherin-1 (PCDH1) as a critical determinant of attachment, entry, and infection by New-World hantaviruses, but not their Old-World counterparts, in primary human microvascular endothelial cells (Jangra et al, 2018), which are major targets of hantavirus infection in vivo (Mackow and Gavrilovskaya, 2009).

Results

Conclusion