PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2

Abstract In search for broad-spectrum antivirals, we discover a small molecule inhibitor, RMC-113, that potently suppresses the replication of multiple RNA viruses including SARS-CoV-2 in human lung organoids. We demonstrate selective inhibition of the lipid kinases PIP4K2C and PIKfyve by RMC-113 an...

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Main Authors: Marwah Karim, Manjari Mishra, Chieh-Wen Lo, Sirle Saul, Halise Busra Cagirici, Manon Gourdelier, Luca Ghita, Amrita Ojha, Do Hoang Nhu Tran, Aditi Agrawal, Connor McGraw, Michael P. East, Karen Anbro Gammeltoft, Malaya Kumar Sahoo, Nancie A. Mooney, Gary L. Johnson, Soumita Das, Pieter Leyssen, Johan Neyts, Winston Chiu, Courtney A. Cohen, Jens Bukh, Judith Gottwein, John M. Dye, Norma Neff, Peter K. Jackson, Benjamin A. Pinsky, Tuomo Laitinen, Tatu Pantsar, Antti Poso, Fabio Zanini, Steven De Jonghe, Christopher R. M. Asquith, Shirit Einav
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61759-1
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author Marwah Karim
Manjari Mishra
Chieh-Wen Lo
Sirle Saul
Halise Busra Cagirici
Manon Gourdelier
Luca Ghita
Amrita Ojha
Do Hoang Nhu Tran
Aditi Agrawal
Connor McGraw
Michael P. East
Karen Anbro Gammeltoft
Malaya Kumar Sahoo
Nancie A. Mooney
Gary L. Johnson
Soumita Das
Pieter Leyssen
Johan Neyts
Winston Chiu
Courtney A. Cohen
Jens Bukh
Judith Gottwein
John M. Dye
Norma Neff
Peter K. Jackson
Benjamin A. Pinsky
Tuomo Laitinen
Tatu Pantsar
Antti Poso
Fabio Zanini
Steven De Jonghe
Christopher R. M. Asquith
Shirit Einav
author_facet Marwah Karim
Manjari Mishra
Chieh-Wen Lo
Sirle Saul
Halise Busra Cagirici
Manon Gourdelier
Luca Ghita
Amrita Ojha
Do Hoang Nhu Tran
Aditi Agrawal
Connor McGraw
Michael P. East
Karen Anbro Gammeltoft
Malaya Kumar Sahoo
Nancie A. Mooney
Gary L. Johnson
Soumita Das
Pieter Leyssen
Johan Neyts
Winston Chiu
Courtney A. Cohen
Jens Bukh
Judith Gottwein
John M. Dye
Norma Neff
Peter K. Jackson
Benjamin A. Pinsky
Tuomo Laitinen
Tatu Pantsar
Antti Poso
Fabio Zanini
Steven De Jonghe
Christopher R. M. Asquith
Shirit Einav
author_sort Marwah Karim
collection DOAJ
description Abstract In search for broad-spectrum antivirals, we discover a small molecule inhibitor, RMC-113, that potently suppresses the replication of multiple RNA viruses including SARS-CoV-2 in human lung organoids. We demonstrate selective inhibition of the lipid kinases PIP4K2C and PIKfyve by RMC-113 and target engagement by its clickable analog. Lipidomics analysis reveals alteration of SARS-CoV-2-induced phosphoinositide signature by RMC-113 and links its antiviral effect with functional PIP4K2C and PIKfyve inhibition. We identify PIP4K2C’s roles in SARS-CoV-2 entry, RNA replication, and assembly/egress, validating it as a druggable antiviral target. Integrating proteomics, single-cell transcriptomics, and functional assays, reveals that PIP4K2C binds SARS-CoV-2 nonstructural protein 6 and regulates virus-induced autophagic flux impairment. Promoting viral protein degradation by reversing autophagic flux impairment is a mechanism of antiviral action of RMC-113. These findings reveal virus-induced autophagy regulation via PIP4K2C, an understudied kinase, and propose dual PIP4K2C and PIKfyve inhibition as a candidate strategy to combat emerging viruses.
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spelling doaj-art-3fb6a03926714188892dca645295f0792025-08-20T04:03:07ZengNature PortfolioNature Communications2041-17232025-07-0116111810.1038/s41467-025-61759-1PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2Marwah Karim0Manjari Mishra1Chieh-Wen Lo2Sirle Saul3Halise Busra Cagirici4Manon Gourdelier5Luca Ghita6Amrita Ojha7Do Hoang Nhu Tran8Aditi Agrawal9Connor McGraw10Michael P. East11Karen Anbro Gammeltoft12Malaya Kumar Sahoo13Nancie A. Mooney14Gary L. Johnson15Soumita Das16Pieter Leyssen17Johan Neyts18Winston Chiu19Courtney A. Cohen20Jens Bukh21Judith Gottwein22John M. Dye23Norma Neff24Peter K. Jackson25Benjamin A. Pinsky26Tuomo Laitinen27Tatu Pantsar28Antti Poso29Fabio Zanini30Steven De Jonghe31Christopher R. M. Asquith32Shirit Einav33Department of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityDepartment of Pharmacology, School of Medicine, University of North Carolina at Chapel HillDepartment of Infectious Diseases, University of CopenhagenDepartment of Pathology, Stanford University School of MedicineBaxter Laboratory, Department of Microbiology & Immunology. Stanford University School of MedicineDepartment of Pharmacology, School of Medicine, University of North Carolina at Chapel HillBiomedical & Nutritional Science, Center for Pathogen Research & Training (CPRT), University of Massachusetts-LowellKU Leuven, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and ChemotherapyKU Leuven, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and ChemotherapyKU Leuven, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and ChemotherapyUS Army Medical Research Institute of Infectious Diseases, Viral Immunology BranchDepartment of Infectious Diseases, University of CopenhagenDepartment of Infectious Diseases, University of CopenhagenUS Army Medical Research Institute of Infectious Diseases, Viral Immunology BranchChan Zuckerberg BiohubBaxter Laboratory, Department of Microbiology & Immunology. Stanford University School of MedicineDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversitySchool of Pharmacy, Faculty of Health Sciences, University of Eastern FinlandSchool of Pharmacy, Faculty of Health Sciences, University of Eastern FinlandSchool of Pharmacy, Faculty of Health Sciences, University of Eastern FinlandSchool of Clinical Medicine, UNSW SydneyKU Leuven, Department of Microbiology, Immunology and Transplantation, Rega Institute for Medical Research, Laboratory of Virology and ChemotherapySchool of Pharmacy, Faculty of Health Sciences, University of Eastern FinlandDepartment of Medicine, Division of Infectious Diseases and Geographic Medicine, Stanford UniversityAbstract In search for broad-spectrum antivirals, we discover a small molecule inhibitor, RMC-113, that potently suppresses the replication of multiple RNA viruses including SARS-CoV-2 in human lung organoids. We demonstrate selective inhibition of the lipid kinases PIP4K2C and PIKfyve by RMC-113 and target engagement by its clickable analog. Lipidomics analysis reveals alteration of SARS-CoV-2-induced phosphoinositide signature by RMC-113 and links its antiviral effect with functional PIP4K2C and PIKfyve inhibition. We identify PIP4K2C’s roles in SARS-CoV-2 entry, RNA replication, and assembly/egress, validating it as a druggable antiviral target. Integrating proteomics, single-cell transcriptomics, and functional assays, reveals that PIP4K2C binds SARS-CoV-2 nonstructural protein 6 and regulates virus-induced autophagic flux impairment. Promoting viral protein degradation by reversing autophagic flux impairment is a mechanism of antiviral action of RMC-113. These findings reveal virus-induced autophagy regulation via PIP4K2C, an understudied kinase, and propose dual PIP4K2C and PIKfyve inhibition as a candidate strategy to combat emerging viruses.https://doi.org/10.1038/s41467-025-61759-1
spellingShingle Marwah Karim
Manjari Mishra
Chieh-Wen Lo
Sirle Saul
Halise Busra Cagirici
Manon Gourdelier
Luca Ghita
Amrita Ojha
Do Hoang Nhu Tran
Aditi Agrawal
Connor McGraw
Michael P. East
Karen Anbro Gammeltoft
Malaya Kumar Sahoo
Nancie A. Mooney
Gary L. Johnson
Soumita Das
Pieter Leyssen
Johan Neyts
Winston Chiu
Courtney A. Cohen
Jens Bukh
Judith Gottwein
John M. Dye
Norma Neff
Peter K. Jackson
Benjamin A. Pinsky
Tuomo Laitinen
Tatu Pantsar
Antti Poso
Fabio Zanini
Steven De Jonghe
Christopher R. M. Asquith
Shirit Einav
PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
Nature Communications
title PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
title_full PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
title_fullStr PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
title_full_unstemmed PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
title_short PIP4K2C inhibition reverses autophagic flux impairment induced by SARS-CoV-2
title_sort pip4k2c inhibition reverses autophagic flux impairment induced by sars cov 2
url https://doi.org/10.1038/s41467-025-61759-1
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