Global functional genomics reveals GRK5 as a cystic fibrosis therapeutic target synergistic with current modulators
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.
Authors
- Miquéias Lopes‐Pacheco (ORCID: https://orcid.org/0000-0002-7444-9359)
- Beate Neumann (ORCID: https://orcid.org/0000-0002-6899-502X)
- Rainer Pepperkok (ORCID: https://orcid.org/0000-0002-9762-3583)
- Jiraporn Ousingsawat (ORCID: https://orcid.org/0000-0003-4318-2601)
- Ines Pankonien (ORCID: https://orcid.org/0000-0002-4622-7521)
- Karl Kunzelmann (ORCID: https://orcid.org/0000-0002-4583-7037)
- Hugo M. Botelho (ORCID: https://orcid.org/0000-0002-4208-1086)
- Violeta Railean (ORCID: https://orcid.org/0000-0003-3713-7089)
- Luka A. Clarke (ORCID: https://orcid.org/0000-0003-3254-9121)
- Vasco Cachatra (ORCID: https://orcid.org/0000-0002-6428-2153)
- Madalena C. Pinto (ORCID: https://orcid.org/0000-0001-9045-269X)
- Margarida D. Amaral (ORCID: https://orcid.org/0000-0002-0828-8630)
- Christian Tischer (ORCID: https://orcid.org/0000-0003-4105-1990)
- Cristina Moiteiro (ORCID: https://orcid.org/0000-0002-9276-6788)
- Mariana F Caleiro
Institutions
- University of Lisbon (PT)
- European Molecular Biology Laboratory (DE)
- European Molecular Biology Laboratory (DE)
- University of Regensburg (DE)
Publication Details
- Journal
- iScience
- Published
- 2025-02-01
- DOI
- https://doi.org/10.1016/j.isci.2025.111942
- Citations
- 3
- Primary Topic
- Cystic Fibrosis Research Advances
- Type
- article
- Field-Weighted Citation Impact
- 4.56
Funders
- Cystic Fibrosis Foundation
- Gilead Sciences
- Gilead Research Scholars
- European Commission
- Fundação para a Ciência e a Tecnologia
- Sixth Framework Programme