Journal article
Novel Cystic Fibrosis Ferret Model Enables Visualization of CFTR Expression Cells and Genetic CFTR Reactivation
Human gene therapy, Vol.36(15-16), pp.1053-1066
08/01/2025
DOI: 10.1089/hum.2024.215
PMCID: PMC12419444
PMID: 39791230
Abstract
Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR). While gene therapy holds promise as a cure, the cell-type-specific heterogeneity of CFTR expression in the lung presents significant challenges. Current CF ferret models closely replicate the human disease phenotype but have limitations in studying functional complementation through cell-type-specific CFTR restoration. To address this, we developed a new transgenic ferret line, CFTRint1-eGFP(lsl), in which a Cre-recombinase (Cre)-excisable enhanced fluorescent protein (eGFP) reporter cassette is knocked in (KI) to intron 1 of the CFTR locus. Breeding this reporter line with CFTRG551D CF ferret resulted in a novel CF model, CFTRint1-eGFP(lsl)/G551D, with disease onset manageable via the administration of CFTR modulator VX770. In this study, we confirmed two key properties of the CFTRint1-eGFP(lsl)/G551D CF ferrets: (1) cell-type-specific expression of the CFTR(N-24)-eGFP fusion protein, driven by the intrinsic CFTR promoter, in polarized epithelial cultures and selected tissues, and (2) functional reversion of the KI allele via Cre-mediated excision of the reporter cassette. This model provides a valuable tool for studying the effects of targeted CFTR reactivation in a cell-type-specific manner, which is crucial for enhancing our understanding of CFTR’s roles in modulating airway clearance and innate immunity, and for identifying relevant cellular targets for CF gene therapy.
Details
- Title: Subtitle
- Novel Cystic Fibrosis Ferret Model Enables Visualization of CFTR Expression Cells and Genetic CFTR Reactivation
- Creators
- Feng Yuan - University of IowaXingshen Sun - University of IowaSoo Yeun Park - University of IowaYinghua Tang - University of IowaZehua Feng - University of IowaMehrnoosh Ebadi - University of IowaYaling Yi - University of IowaAdriane E Thompson - Department of Anatomy and Cell Biology, Carver College of Medicine, University of Iowa, Iowa City, Iowa, USAJoseph D Karippaparambil - University of IowaJohn F Engelhardt - University of IowaZiying Yan - University of Iowa
- Resource Type
- Journal article
- Publication Details
- Human gene therapy, Vol.36(15-16), pp.1053-1066
- DOI
- 10.1089/hum.2024.215
- PMID
- 39791230
- PMCID
- PMC12419444
- NLM abbreviation
- Hum Gene Ther
- ISSN
- 1043-0342
- eISSN
- 1557-7422
- Publisher
- MARY ANN LIEBERT, INC; NEW ROCHELLE
- Grant note
- National Institutes of Health: R01 HL174593, R21 AI182645, P30 DK054759, RC2 DK124207, P01 HL152960, R01 HL165404, 75N92024C00008 Cystic Fibrosis Foundation: YAN23G0, ENGELH21XX0, 004590G222 Cystic Fibrosis Research Institute: 1267026
This work was funded by the following grants from the National Institutes of Health (R01 HL174593, R21 AI182645 to ZY; P30 DK054759, RC2 DK124207, P01 HL152960, and R01 HL165404 and Federal Contract 75N92024C00008 to J.F.E.), the Cystic Fibrosis Foundation (YAN23G0 to Z.Y.; ENGELH21XX0 to J.F.E.; and 004590G222 to X.S.), and the Cystic Fibrosis Research Institute (Grant#1267026 to F.Y.).
- Language
- English
- Electronic publication date
- 01/10/2025
- Date published
- 08/01/2025
- Academic Unit
- Anatomy and Cell Biology; Radiation Oncology; Internal Medicine
- Record Identifier
- 9984771631402771
Metrics
6 Record Views