Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators

Summary: Cystic fibrosis (CF) is a life-shortening disease affecting >160,000 individuals worldwide predominantly with respiratory symptoms. About 80% of individuals with CF have the p.Phe508del variant that causes the CF transmembrane conductance regulator (CFTR) protein to misfold and be target...

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Main Authors: Hugo M. Botelho, Miquéias Lopes-Pacheco, Madalena C. Pinto, Violeta Railean, Ines Pankonien, Mariana F. Caleiro, Luka A. Clarke, Vasco Cachatra, Beate Neumann, Christian Tischer, Cristina Moiteiro, Jiraporn Ousingsawat, Karl Kunzelmann, Rainer Pepperkok, Margarida D. Amaral
Format: Article
Language:English
Published: Elsevier 2025-03-01
Series:iScience
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Online Access:http://www.sciencedirect.com/science/article/pii/S2589004225002020
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author Hugo M. Botelho
Miquéias Lopes-Pacheco
Madalena C. Pinto
Violeta Railean
Ines Pankonien
Mariana F. Caleiro
Luka A. Clarke
Vasco Cachatra
Beate Neumann
Christian Tischer
Cristina Moiteiro
Jiraporn Ousingsawat
Karl Kunzelmann
Rainer Pepperkok
Margarida D. Amaral
author_facet Hugo M. Botelho
Miquéias Lopes-Pacheco
Madalena C. Pinto
Violeta Railean
Ines Pankonien
Mariana F. Caleiro
Luka A. Clarke
Vasco Cachatra
Beate Neumann
Christian Tischer
Cristina Moiteiro
Jiraporn Ousingsawat
Karl Kunzelmann
Rainer Pepperkok
Margarida D. Amaral
author_sort Hugo M. Botelho
collection DOAJ
description Summary: Cystic fibrosis (CF) is a life-shortening disease affecting >160,000 individuals worldwide predominantly with respiratory symptoms. About 80% of individuals with CF have the p.Phe508del variant that causes the CF transmembrane conductance regulator (CFTR) protein to misfold and be targeted for premature degradation by the endoplasmic reticulum (ER) quality control (ERQC), thus preventing its plasma membrane (PM) traffic. Despite the recent approval of a “highly effective” drug rescuing p.Phe508del-CFTR, maximal lung function improvement is ∼14%. To identify global modulators of p.Phe508del traffic, we performed a high-content small interfering RNA (siRNA) microscopy-based screen of >9,000 genes and monitored p.Phe508del-CFTR PM rescue in human airway cells. This primary screen identified 227 p.Phe508del-CFTR traffic regulators, of which 35 could be validated by additional siRNAs. Subsequent mechanistic studies established GRK5 as a robust regulator whose inhibition rescues p.Phe508del-CFTR PM traffic and function in primary and immortalized cells, thus emerging as a novel potential drug target for CF.
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spelling doaj-art-2bc0cff47acc422e867718e3b48d78802025-08-20T03:11:46ZengElsevieriScience2589-00422025-03-0128311194210.1016/j.isci.2025.111942Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulatorsHugo M. Botelho0Miquéias Lopes-Pacheco1Madalena C. Pinto2Violeta Railean3Ines Pankonien4Mariana F. Caleiro5Luka A. Clarke6Vasco Cachatra7Beate Neumann8Christian Tischer9Cristina Moiteiro10Jiraporn Ousingsawat11Karl Kunzelmann12Rainer Pepperkok13Margarida D. Amaral14BioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, Portugal; Department of Physiology, University of Regensburg, Universitätsstrasse 31, 93053 Regensburg, GermanyBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalCentro de Química Estrutural, Institute of Molecular Sciences, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalCell Biology and Biophysics Unit and Advanced Light Microscopy Facility, European Molecular Biology Laboratory (EMBL), Meyerhofstraße 1, 69117 Heidelberg, GermanyCell Biology and Biophysics Unit and Advanced Light Microscopy Facility, European Molecular Biology Laboratory (EMBL), Meyerhofstraße 1, 69117 Heidelberg, Germany; Centre for Bioimage Analysis, European Molecular Biology Laboratory (EMBL), Meyerhofstraße 1, 69117 Heidelberg, GermanyCentro de Química Estrutural, Institute of Molecular Sciences, Department of Chemistry and Biochemistry, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, PortugalDepartment of Physiology, University of Regensburg, Universitätsstrasse 31, 93053 Regensburg, GermanyDepartment of Physiology, University of Regensburg, Universitätsstrasse 31, 93053 Regensburg, GermanyCell Biology and Biophysics Unit and Advanced Light Microscopy Facility, European Molecular Biology Laboratory (EMBL), Meyerhofstraße 1, 69117 Heidelberg, GermanyBioISI – Biosystems & Integrative Sciences Institute, Faculty of Sciences, University of Lisboa, Campo Grande, 1749-016 Lisboa, Portugal; Corresponding authorSummary: Cystic fibrosis (CF) is a life-shortening disease affecting >160,000 individuals worldwide predominantly with respiratory symptoms. About 80% of individuals with CF have the p.Phe508del variant that causes the CF transmembrane conductance regulator (CFTR) protein to misfold and be targeted for premature degradation by the endoplasmic reticulum (ER) quality control (ERQC), thus preventing its plasma membrane (PM) traffic. Despite the recent approval of a “highly effective” drug rescuing p.Phe508del-CFTR, maximal lung function improvement is ∼14%. To identify global modulators of p.Phe508del traffic, we performed a high-content small interfering RNA (siRNA) microscopy-based screen of >9,000 genes and monitored p.Phe508del-CFTR PM rescue in human airway cells. This primary screen identified 227 p.Phe508del-CFTR traffic regulators, of which 35 could be validated by additional siRNAs. Subsequent mechanistic studies established GRK5 as a robust regulator whose inhibition rescues p.Phe508del-CFTR PM traffic and function in primary and immortalized cells, thus emerging as a novel potential drug target for CF.http://www.sciencedirect.com/science/article/pii/S2589004225002020Cell biologyFunctional genomics
spellingShingle Hugo M. Botelho
Miquéias Lopes-Pacheco
Madalena C. Pinto
Violeta Railean
Ines Pankonien
Mariana F. Caleiro
Luka A. Clarke
Vasco Cachatra
Beate Neumann
Christian Tischer
Cristina Moiteiro
Jiraporn Ousingsawat
Karl Kunzelmann
Rainer Pepperkok
Margarida D. Amaral
Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
iScience
Cell biology
Functional genomics
title Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
title_full Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
title_fullStr Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
title_full_unstemmed Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
title_short Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
title_sort global functional genomics reveals grk5 as a cystic fibrosis therapeutic target synergistic with current modulators
topic Cell biology
Functional genomics
url http://www.sciencedirect.com/science/article/pii/S2589004225002020
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