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Highly efficient genome editing in primary human bronchial epithelial cells differentiated at air-liquid interface

Radu Rapiteanu, Tina Karagyozova, Natalie Zimmermann, Kuljit Singh, Gareth Wayne, Matteo Martufi, Nikolai N. Belyaev, Edith M. Hessel, David Michalovich, Ricardo Macarron, Wendy C. Rowan, William J. Cairns, Jan Roger, Joanna Betts, Soren Beinke, Klio Maratou
European Respiratory Journal 2020; DOI: 10.1183/13993003.00950-2019
Radu Rapiteanu
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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  • ORCID record for Radu Rapiteanu
  • For correspondence: radu.a.rapiteanu@gsk.com
Tina Karagyozova
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Natalie Zimmermann
2Refractory Respiratory Inflammation DPU, GlaxoSmithKline R&D, Stevenage, UK
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Kuljit Singh
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Gareth Wayne
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Matteo Martufi
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Nikolai N. Belyaev
2Refractory Respiratory Inflammation DPU, GlaxoSmithKline R&D, Stevenage, UK
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Edith M. Hessel
2Refractory Respiratory Inflammation DPU, GlaxoSmithKline R&D, Stevenage, UK
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David Michalovich
2Refractory Respiratory Inflammation DPU, GlaxoSmithKline R&D, Stevenage, UK
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Ricardo Macarron
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Wendy C. Rowan
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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William J. Cairns
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Jan Roger
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Joanna Betts
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Soren Beinke
2Refractory Respiratory Inflammation DPU, GlaxoSmithKline R&D, Stevenage, UK
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Klio Maratou
1Target Sciences, GlaxoSmithKline R&D, Stevenage, UK
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Extract

The structure and composition of the bronchial epithelium is altered in respiratory diseases such as COPD and asthma, in which goblet cell hyperplasia and reduced numbers of ciliated cells impair mucociliary clearance. Current standard of care focuses on alleviating disease symptoms such as airway obstruction and inflammation using bronchodilators or steroids, respectively. Whilst these approaches are effective, they do not address the underlying pathogenic processes. An alternative way forward would be to identify and target the dysregulated cellular pathways responsible for impaired bronchial epithelial function. In this study we describe a single-step, highly efficient and easily scalable genome editing pipeline to aid the dissection of the molecular mechanisms underlying primary human bronchial epithelial cell (BEC) differentiation and function at air liquid interface (ALI).

Footnotes

This manuscript has recently been accepted for publication in the European Respiratory Journal. It is published here in its accepted form prior to copyediting and typesetting by our production team. After these production processes are complete and the authors have approved the resulting proofs, the article will move to the latest issue of the ERJ online. Please open or download the PDF to view this article.

Conflict of interest: Dr. Rapiteanu has nothing to disclose.

Conflict of interest: Ms Karagyozova has nothing to disclose.

Conflict of interest: Dr. Zimmermann has nothing to disclose.

Conflict of interest: Dr. Singh has nothing to disclose.

Conflict of interest: Dr. Wayne has nothing to disclose.

Conflict of interest: Dr. Martufi has nothing to disclose.

Conflict of interest: Dr. Belyaev has nothing to disclose.

Conflict of interest: Dr. Hessel reports other from GlaxoSmithKline, during the conduct of the study.

Conflict of interest: Dr. Michalovich reports I am an employee of GSK and shareholder.

Conflict of interest: Dr. Macarron has nothing to disclose.

Conflict of interest: Dr. Rowan has nothing to disclose.

Conflict of interest: Dr. Cairns has nothing to disclose.

Conflict of interest: Dr. Roger has nothing to disclose.

Conflict of interest: Dr. Betts has nothing to disclose.

Conflict of interest: Dr. Beinke has nothing to disclose.

Conflict of interest: Dr. Maratou has nothing to disclose.

  • Received February 1, 2019.
  • Accepted January 31, 2020.
  • Copyright ©ERS 2020
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Highly efficient genome editing in primary human bronchial epithelial cells differentiated at air-liquid interface
Radu Rapiteanu, Tina Karagyozova, Natalie Zimmermann, Kuljit Singh, Gareth Wayne, Matteo Martufi, Nikolai N. Belyaev, Edith M. Hessel, David Michalovich, Ricardo Macarron, Wendy C. Rowan, William J. Cairns, Jan Roger, Joanna Betts, Soren Beinke, Klio Maratou
European Respiratory Journal Jan 2020, 1900950; DOI: 10.1183/13993003.00950-2019

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Highly efficient genome editing in primary human bronchial epithelial cells differentiated at air-liquid interface
Radu Rapiteanu, Tina Karagyozova, Natalie Zimmermann, Kuljit Singh, Gareth Wayne, Matteo Martufi, Nikolai N. Belyaev, Edith M. Hessel, David Michalovich, Ricardo Macarron, Wendy C. Rowan, William J. Cairns, Jan Roger, Joanna Betts, Soren Beinke, Klio Maratou
European Respiratory Journal Jan 2020, 1900950; DOI: 10.1183/13993003.00950-2019
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