SARS-CoV-2 infects an upper airway model derived from induced pluripotent stem cells

Ivo Djidrovski; Maria Georgiou; Grant L. Hughes; Edward I. Patterson; Aitor Casas-Sanchez; Shaun H. Pennington; Giancarlo A. Biagini; Marina Moya-Molina; Jelle van den Bor; Martine J. Smit; Git Chung; Majlinda Lako; Lyle Armstrong

doi:10.1002/stem.3422

SARS-CoV-2 infects an upper airway model derived from induced pluripotent stem cells

Ivo Djidrovski, Maria Georgiou, Grant L. Hughes, Edward I. Patterson, Aitor Casas-Sanchez, Shaun H. Pennington, Giancarlo A. Biagini, Marina Moya-Molina, Jelle van den Bor, Martine J. Smit, Git Chung, Majlinda Lako, Lyle Armstrong^*

^*Corresponding author for this work

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

Abstract

As one of the primary points of entry of xenobiotic substances and infectious agents into the body, the lungs are subject to a range of dysfunctions and diseases that together account for a significant number of patient deaths. In view of this, there is an outstanding need for in vitro systems in which to assess the impact of both infectious agents and xenobiotic substances of the lungs. To address this issue, we have developed a protocol to generate airway epithelial basal-like cells from induced pluripotent stem cells, which simplifies the manufacture of cellular models of the human upper airways. Basal-like cells generated in this study were cultured on transwell inserts to allow formation of a confluent monolayer and then exposed to an air-liquid interface to induce differentiation into a pseudostratified epithelial construct with a marked similarity to the upper airway epithelium in vivo. These constructs contain the component cell types required of an epithelial model system, produce mucus and functional cilia, and can support SARS-CoV-2 infection/replication and the secretion of cytokines in a manner similar to that of in vivo airways. This method offers a readily accessible and highly scalable protocol for the manufacture of upper airway models that could find applications in development of therapies for respiratory viral infections and the assessment of drug toxicity on the human lungs.

Original language	English
Pages (from-to)	1310-1321
Number of pages	12
Journal	Stem Cells
Volume	39
Issue number	10
Early online date	21 Jun 2021
DOIs	https://doi.org/10.1002/stem.3422
Publication status	Published - Oct 2021

Bibliographical note

Funding Information:
Financial support from the Marie Sk?odowska-Curie Action - Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC?Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI-BBSRC COVID rolling fund (BB/V017772/1).

Funding Information:
Financial support from the Marie Skłodowska‐Curie Action ‐ Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC—Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI‐BBSRC COVID rolling fund (BB/V017772/1).

Publisher Copyright:
© 2021 The Authors. STEM CELLS published by Wiley Periodicals LLC on behalf of AlphaMed Press.

Copyright:
Copyright 2021 Elsevier B.V., All rights reserved.

Funding

Financial support from the Marie Sk?odowska-Curie Action - Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC?Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI-BBSRC COVID rolling fund (BB/V017772/1). Financial support from the Marie Skłodowska‐Curie Action ‐ Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC—Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI‐BBSRC COVID rolling fund (BB/V017772/1).

Funders	Funder number
Biotechnology and Biosciences Research Council	BB/V01126X/1
Department of Health or Public Health England
Liverpool School of Tropical Medicine Director's Catalyst Fund	V011278/1, BB/T001240/1
UKRI-BBSRC
UKRI‐BBSRC	BB/V017772/1
URKI	20197, NIHR2000907
National Institute of Allergy and Infectious Diseases	R21AI138074
H2020 Marie Skłodowska-Curie Actions	721975, IN3
UK Research and Innovation	SIPF 20197
National Institute for Health Research Health Protection Research Unit
Medical Research Council	MR/836 S00467X/1
Royal Society	RSWF\R1\180013
Oxford University
Public Health England

Keywords

cytokines
induced pluripotent stem cells
interleukins
lung

UN SDGs

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

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10.1002/stem.3422

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Cite this

@article{cc68be63843241a8831e704dca41226d,

title = "SARS-CoV-2 infects an upper airway model derived from induced pluripotent stem cells",

abstract = "As one of the primary points of entry of xenobiotic substances and infectious agents into the body, the lungs are subject to a range of dysfunctions and diseases that together account for a significant number of patient deaths. In view of this, there is an outstanding need for in vitro systems in which to assess the impact of both infectious agents and xenobiotic substances of the lungs. To address this issue, we have developed a protocol to generate airway epithelial basal-like cells from induced pluripotent stem cells, which simplifies the manufacture of cellular models of the human upper airways. Basal-like cells generated in this study were cultured on transwell inserts to allow formation of a confluent monolayer and then exposed to an air-liquid interface to induce differentiation into a pseudostratified epithelial construct with a marked similarity to the upper airway epithelium in vivo. These constructs contain the component cell types required of an epithelial model system, produce mucus and functional cilia, and can support SARS-CoV-2 infection/replication and the secretion of cytokines in a manner similar to that of in vivo airways. This method offers a readily accessible and highly scalable protocol for the manufacture of upper airway models that could find applications in development of therapies for respiratory viral infections and the assessment of drug toxicity on the human lungs.",

keywords = "cytokines, induced pluripotent stem cells, interleukins, lung",

author = "Ivo Djidrovski and Maria Georgiou and Hughes, {Grant L.} and Patterson, {Edward I.} and Aitor Casas-Sanchez and Pennington, {Shaun H.} and Biagini, {Giancarlo A.} and Marina Moya-Molina and {van den Bor}, Jelle and Smit, {Martine J.} and Git Chung and Majlinda Lako and Lyle Armstrong",

note = "Funding Information: Financial support from the Marie Sk?odowska-Curie Action - Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC?Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI-BBSRC COVID rolling fund (BB/V017772/1). Funding Information: Financial support from the Marie Sk{\l}odowska‐Curie Action ‐ Innovative Training Network, grant number 721975 (IN3) and grant funding from the Biotechnology and Biosciences Research Council (BBSRC) (grant reference BB/V01126X/1) held by Majlinda Lako and colleagues. E.I.P. was supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund award. G.L.H. was supported by the BBSRC (BB/T001240/1 and V011278/1), a Royal Society Wolfson Fellowship (RSWF\R1\180013), the NIH (R21AI138074), URKI (20197), and the NIHR (NIHR2000907). G.L.H. is affiliated to the National Institute for Health Research Health Protection Research Unit (NIHR HPRU) in Emerging and Zoonotic Infections at University of Liverpool in partnership with Public Health England (PHE), in collaboration with Liverpool School of Tropical Medicine and the University of Oxford. G.L.H. is based at LSTM. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, the Department of Health or Public Health England. We also thank the MCCF (Advanced Microscopy and Cytometry Research Core Facility of the Amsterdam UMC—Location VU University Medical Center) for imaging support. G.A.B. and S.H.P. acknowledge support from Medical Research Council (MR/836 S00467X/1) and the UK Research and Innovation (UKRI) Strength in Places Fund (SIPF 20197). A.C.S. was supported by supported by the Liverpool School of Tropical Medicine Director's Catalyst Fund and the UKRI‐BBSRC COVID rolling fund (BB/V017772/1). Publisher Copyright: {\textcopyright} 2021 The Authors. STEM CELLS published by Wiley Periodicals LLC on behalf of AlphaMed Press. Copyright: Copyright 2021 Elsevier B.V., All rights reserved.",

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month = oct,

doi = "10.1002/stem.3422",

language = "English",

volume = "39",

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AU - Djidrovski, Ivo

AU - Georgiou, Maria

AU - Hughes, Grant L.

AU - Patterson, Edward I.

AU - Casas-Sanchez, Aitor

AU - Pennington, Shaun H.

AU - Biagini, Giancarlo A.

AU - Moya-Molina, Marina

AU - van den Bor, Jelle

AU - Smit, Martine J.

AU - Chung, Git

AU - Lako, Majlinda

AU - Armstrong, Lyle

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N2 - As one of the primary points of entry of xenobiotic substances and infectious agents into the body, the lungs are subject to a range of dysfunctions and diseases that together account for a significant number of patient deaths. In view of this, there is an outstanding need for in vitro systems in which to assess the impact of both infectious agents and xenobiotic substances of the lungs. To address this issue, we have developed a protocol to generate airway epithelial basal-like cells from induced pluripotent stem cells, which simplifies the manufacture of cellular models of the human upper airways. Basal-like cells generated in this study were cultured on transwell inserts to allow formation of a confluent monolayer and then exposed to an air-liquid interface to induce differentiation into a pseudostratified epithelial construct with a marked similarity to the upper airway epithelium in vivo. These constructs contain the component cell types required of an epithelial model system, produce mucus and functional cilia, and can support SARS-CoV-2 infection/replication and the secretion of cytokines in a manner similar to that of in vivo airways. This method offers a readily accessible and highly scalable protocol for the manufacture of upper airway models that could find applications in development of therapies for respiratory viral infections and the assessment of drug toxicity on the human lungs.

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SARS-CoV-2 infects an upper airway model derived from induced pluripotent stem cells

Abstract

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UN SDGs

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