MFSD6 is an entry receptor for enterovirus D68

Lauren Varanese; Lily Xu; Christine E Peters; Grigore Pintilie; David S Roberts; Suyash Raj; Mengying Liu; Yaw Shin Ooi; Jonathan Diep; Wenjie Qiao; Christopher M Richards; Jeremy Callaway; Carolyn R Bertozzi; Sabrina Jabs; Erik de Vries; Frank J M van Kuppeveld; Claude M Nagamine; Wah Chiu; Jan E Carette

doi:10.1038/s41586-025-08908-0

MFSD6 is an entry receptor for enterovirus D68

Lauren Varanese
, Lily Xu
, Christine E Peters
, Grigore Pintilie
, David S Roberts
, Suyash Raj
, Mengying Liu
, Yaw Shin Ooi
, Jonathan Diep
, Wenjie Qiao
, Christopher M Richards
, Jeremy Callaway
, Carolyn R Bertozzi
, Sabrina Jabs
, Erik de Vries
, Frank J M van Kuppeveld
, Claude M Nagamine
, Wah Chiu^*
, Jan E Carette^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

With the near eradication of poliovirus due to global vaccination campaigns, attention has shifted to other enteroviruses that can cause polio-like paralysis syndrome (now termed acute flaccid myelitis)^{1, 2–3}. In particular, enterovirus D68 (EV-D68) is believed to be the main driver of epidemic outbreaks of acute flaccid myelitis in recent years⁴, yet not much is known about EV-D68 host interactions. EV-D68 is a respiratory virus⁵ but, in rare cases, can spread to the central nervous system to cause severe neuropathogenesis. Here we use genome-scale CRISPR screens to identify the poorly characterized multipass membrane transporter MFSD6 as a host entry factor for EV-D68. Knockout of MFSD6 expression abrogated EV-D68 infection in cell lines and primary cells corresponding to respiratory and neural cells. MFSD6 localized to the plasma membrane and was required for viral entry into host cells. MFSD6 bound directly to EV-D68 particles through its extracellular, third loop (L3). We determined the cryo-electron microscopy structure of EV-D68 in a complex with MFSD6 L3, revealing the interaction interface. A decoy receptor, engineered by fusing MFSD6 L3 to Fc, blocked EV-D68 infection of human primary lung epithelial cells and provided near-complete protection in a lethal mouse model of EV-D68 infection. Collectively, our results reveal MFSD6 as an entry receptor for EV-D68, and support the targeting of MFSD6 as a potential mechanism to combat infections by this emerging pathogen with pandemic potential.

Original language	English
Pages (from-to)	1268-1275
Number of pages	8
Journal	Nature
Volume	641
Issue number	8065
Early online date	25 Mar 2025
DOIs	https://doi.org/10.1038/s41586-025-08908-0
Publication status	Published - 29 May 2025

Bibliographical note

Publisher Copyright:
© The Author(s), under exclusive licence to Springer Nature Limited 2025.

Funding

We thank the members of J.E.C.\u2019s laboratory and the members of W.C.\u2019s laboratory, especially A. Dupzyk, B. Waldman and F. Cooper, for their feedback and insights; I. L. Weissman, G. Tender and C. Xiao for their advice and feedback; M. Amieva and D. Monack for the use of their confocal microscope; D. Monack for the use of her tissue homogenizer; the members of the ISCRBM FACS Core for the use of their Sony MA900 system. Schematics in Figs. and were created using BioRender. This work was funded in part by the National Institutes of Health (NIH) T32G Stanford Cellular and Molecular Biology Training Program M007276 (to L.V.); NIH T32 AI00732 (to C.E.P.); NIH R01 CA200423 (to C.R.B.); NIH R01 AI169467 (to J.E.C.); NIH R01 AI153169 (to J.E.C.); NIH R01 AI140186 (to J.E.C.); National Science Foundation (NSF) GRFP DGE-1656518 (to L.X.), NSF DGE-1656518 (to C.E.P.) and NSF DGE-2146755 (to S.R.); Damon Runyon Cancer Research Foundation DRG-2526-24 (to D.S.R.); Burroughs Wellcome Fund Investigators in the Pathogenesis of Infectious Disease (to J.E.C.); Stanford Maternal & Child Health Research Institute (to W.Q.); European Union\u2019s Horizon Research and Innovation program PANVIPREP (grant number 101003627) (to F.J.M.v.K.); and EU Horizon ERC Advanced Grant program VIRLUMINOUS (grant number 01053576) (to F.J.M.v.K). Cell sorting was done through the Stanford Shared FACS Facility (RRID: SCR_017788) using either the BD FACSAria II (funded by NIH S10RR025518-01) or BD FACSFusion (purchased by the Parker Institute for Cancer Immunology) sorter. Cryo-EM data were collected at the Stanford-SLAC CryoEM Center (supported by the NIH Common Fund Transformative High-Resolution Cryo-Electron Microscopy program U24GM129541 to W.C.).

Funders	Funder number
Stanford Maternal and Child Health Research Institute
European Union’s Horizon Research and Innovation program PANVIPREP	101003627
National Science Foundation	DGE-1656518, DGE-2146755, GRFP DGE-1656518
Damon Runyon Cancer Research Foundation	DRG-2526-24
National Institutes of Health	T32 AI00732, M007276, R01 AI153169, R01 CA200423, R01 AI140186, R01 AI169467
EU Horizon ERC	SCR_017788, 01053576, S10RR025518-01
Parker Institute for Cancer Immunology	U24GM129541

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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s41586-025-08908-0Final published version, 49.2 MBLicence: Taverne

Cite this

Varanese, L., Xu, L., Peters, C. E., Pintilie, G., Roberts, D. S., Raj, S., Liu, M., Ooi, Y. S., Diep, J., Qiao, W., Richards, C. M., Callaway, J., Bertozzi, C. R., Jabs, S., de Vries, E., van Kuppeveld, F. J. M., Nagamine, C. M., Chiu, W., & Carette, J. E. (2025). MFSD6 is an entry receptor for enterovirus D68. Nature, 641(8065), 1268-1275. https://doi.org/10.1038/s41586-025-08908-0

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title = "MFSD6 is an entry receptor for enterovirus D68",

abstract = "With the near eradication of poliovirus due to global vaccination campaigns, attention has shifted to other enteroviruses that can cause polio-like paralysis syndrome (now termed acute flaccid myelitis)1, 2–3. In particular, enterovirus D68 (EV-D68) is believed to be the main driver of epidemic outbreaks of acute flaccid myelitis in recent years4, yet not much is known about EV-D68 host interactions. EV-D68 is a respiratory virus5 but, in rare cases, can spread to the central nervous system to cause severe neuropathogenesis. Here we use genome-scale CRISPR screens to identify the poorly characterized multipass membrane transporter MFSD6 as a host entry factor for EV-D68. Knockout of MFSD6 expression abrogated EV-D68 infection in cell lines and primary cells corresponding to respiratory and neural cells. MFSD6 localized to the plasma membrane and was required for viral entry into host cells. MFSD6 bound directly to EV-D68 particles through its extracellular, third loop (L3). We determined the cryo-electron microscopy structure of EV-D68 in a complex with MFSD6 L3, revealing the interaction interface. A decoy receptor, engineered by fusing MFSD6 L3 to Fc, blocked EV-D68 infection of human primary lung epithelial cells and provided near-complete protection in a lethal mouse model of EV-D68 infection. Collectively, our results reveal MFSD6 as an entry receptor for EV-D68, and support the targeting of MFSD6 as a potential mechanism to combat infections by this emerging pathogen with pandemic potential.",

author = "Lauren Varanese and Lily Xu and Peters, \{Christine E\} and Grigore Pintilie and Roberts, \{David S\} and Suyash Raj and Mengying Liu and Ooi, \{Yaw Shin\} and Jonathan Diep and Wenjie Qiao and Richards, \{Christopher M\} and Jeremy Callaway and Bertozzi, \{Carolyn R\} and Sabrina Jabs and \{de Vries\}, Erik and \{van Kuppeveld\}, \{Frank J M\} and Nagamine, \{Claude M\} and Wah Chiu and Carette, \{Jan E\}",

note = "Publisher Copyright: {\textcopyright} The Author(s), under exclusive licence to Springer Nature Limited 2025.",

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doi = "10.1038/s41586-025-08908-0",

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T1 - MFSD6 is an entry receptor for enterovirus D68

AU - Varanese, Lauren

AU - Xu, Lily

AU - Peters, Christine E

AU - Pintilie, Grigore

AU - Roberts, David S

AU - Raj, Suyash

AU - Liu, Mengying

AU - Ooi, Yaw Shin

AU - Diep, Jonathan

AU - Qiao, Wenjie

AU - Richards, Christopher M

AU - Callaway, Jeremy

AU - Bertozzi, Carolyn R

AU - Jabs, Sabrina

AU - de Vries, Erik

AU - van Kuppeveld, Frank J M

AU - Nagamine, Claude M

AU - Chiu, Wah

AU - Carette, Jan E

PY - 2025/5/29

Y1 - 2025/5/29

N2 - With the near eradication of poliovirus due to global vaccination campaigns, attention has shifted to other enteroviruses that can cause polio-like paralysis syndrome (now termed acute flaccid myelitis)1, 2–3. In particular, enterovirus D68 (EV-D68) is believed to be the main driver of epidemic outbreaks of acute flaccid myelitis in recent years4, yet not much is known about EV-D68 host interactions. EV-D68 is a respiratory virus5 but, in rare cases, can spread to the central nervous system to cause severe neuropathogenesis. Here we use genome-scale CRISPR screens to identify the poorly characterized multipass membrane transporter MFSD6 as a host entry factor for EV-D68. Knockout of MFSD6 expression abrogated EV-D68 infection in cell lines and primary cells corresponding to respiratory and neural cells. MFSD6 localized to the plasma membrane and was required for viral entry into host cells. MFSD6 bound directly to EV-D68 particles through its extracellular, third loop (L3). We determined the cryo-electron microscopy structure of EV-D68 in a complex with MFSD6 L3, revealing the interaction interface. A decoy receptor, engineered by fusing MFSD6 L3 to Fc, blocked EV-D68 infection of human primary lung epithelial cells and provided near-complete protection in a lethal mouse model of EV-D68 infection. Collectively, our results reveal MFSD6 as an entry receptor for EV-D68, and support the targeting of MFSD6 as a potential mechanism to combat infections by this emerging pathogen with pandemic potential.

AB - With the near eradication of poliovirus due to global vaccination campaigns, attention has shifted to other enteroviruses that can cause polio-like paralysis syndrome (now termed acute flaccid myelitis)1, 2–3. In particular, enterovirus D68 (EV-D68) is believed to be the main driver of epidemic outbreaks of acute flaccid myelitis in recent years4, yet not much is known about EV-D68 host interactions. EV-D68 is a respiratory virus5 but, in rare cases, can spread to the central nervous system to cause severe neuropathogenesis. Here we use genome-scale CRISPR screens to identify the poorly characterized multipass membrane transporter MFSD6 as a host entry factor for EV-D68. Knockout of MFSD6 expression abrogated EV-D68 infection in cell lines and primary cells corresponding to respiratory and neural cells. MFSD6 localized to the plasma membrane and was required for viral entry into host cells. MFSD6 bound directly to EV-D68 particles through its extracellular, third loop (L3). We determined the cryo-electron microscopy structure of EV-D68 in a complex with MFSD6 L3, revealing the interaction interface. A decoy receptor, engineered by fusing MFSD6 L3 to Fc, blocked EV-D68 infection of human primary lung epithelial cells and provided near-complete protection in a lethal mouse model of EV-D68 infection. Collectively, our results reveal MFSD6 as an entry receptor for EV-D68, and support the targeting of MFSD6 as a potential mechanism to combat infections by this emerging pathogen with pandemic potential.

UR - https://www.scopus.com/pages/publications/105003978850

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SP - 1268

EP - 1275

JO - Nature

JF - Nature

IS - 8065

ER -

MFSD6 is an entry receptor for enterovirus D68

Abstract

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Funding

UN SDGs

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