Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth

Abstract Background The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage ar...

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Main Authors: Shih-Hsing Leir, Svyatoslav Tkachenko, Alekh Paranjapye, Arnaud J. Van Wettere, Jenny L. Kerschner, Iuri Viotti Perisse, Cheyenne M. Marriott, Tayler Patrick, Ying Liu, Kenneth L. White, Irina A. Polejaeva, Ann Harris
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Molecular Medicine
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Online Access:https://doi.org/10.1186/s10020-025-01266-7
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author Shih-Hsing Leir
Svyatoslav Tkachenko
Alekh Paranjapye
Arnaud J. Van Wettere
Jenny L. Kerschner
Iuri Viotti Perisse
Cheyenne M. Marriott
Tayler Patrick
Ying Liu
Kenneth L. White
Irina A. Polejaeva
Ann Harris
author_facet Shih-Hsing Leir
Svyatoslav Tkachenko
Alekh Paranjapye
Arnaud J. Van Wettere
Jenny L. Kerschner
Iuri Viotti Perisse
Cheyenne M. Marriott
Tayler Patrick
Ying Liu
Kenneth L. White
Irina A. Polejaeva
Ann Harris
author_sort Shih-Hsing Leir
collection DOAJ
description Abstract Background The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage are still unclear. Inflammation may occur in the CF lung prior to birth in human and several large animal models suggesting an in utero origin for the disease and encouraging further studies prior to birth. Methods Here we used the sheep model of CF (CFTR −/− ) and age-matched wild-type (WT) sheep of the same breed to investigate the single cell transcriptomes of proximal and distal lung tissue at 80 days and 120 days of gestation and at term (147 days). Single cell RNA-seq was performed on tissues from 4 to 7 animals of each genotype (WT and CFTR −/− ) at each time point. Results At term, FOXJ1-expressing ciliated cells are overrepresented in both lung regions from CFTR −/− lambs, while secretory epithelial and basal cells are underrepresented in proximal lung, as are T cells and monocytes in distal lung. The imbalance in ciliated and basal cells was confirmed by immunohistochemistry. At 120 days of gestation, lymphoid cells are slightly more abundant in proximal and distal lung from CFTR −/− animals compared to WT, consistent with the transient CF-associated inflammatory response in utero. At 80 days of gestation, T and B cells are underrepresented in both lung regions. Conclusions The differences in epithelial cell abundance observed in the CFTR −/− lambs at term may reflect sequelae from the loss of CFTR on lung development and differentiation in utero. These findings provide novel insights into the cellular mechanisms of pathology and may be relevant to the design of new therapeutic approaches for CF lung disease.
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spelling doaj-art-14a4d68cf4564e3494db6026b181a9fa2025-08-20T03:21:03ZengBMCMolecular Medicine1528-36582025-06-0131112410.1186/s10020-025-01266-7Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birthShih-Hsing Leir0Svyatoslav Tkachenko1Alekh Paranjapye2Arnaud J. Van Wettere3Jenny L. Kerschner4Iuri Viotti Perisse5Cheyenne M. Marriott6Tayler Patrick7Ying Liu8Kenneth L. White9Irina A. Polejaeva10Ann Harris11Department of Genetics and Genome Sciences, Case Western Reserve University School of MedicineDepartment of Genetics and Genome Sciences, Case Western Reserve University School of MedicineDepartment of Genetics and Genome Sciences, Case Western Reserve University School of MedicineDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Genetics and Genome Sciences, Case Western Reserve University School of MedicineDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Animal, Dairy and Veterinary Sciences, Utah State UniversityDepartment of Genetics and Genome Sciences, Case Western Reserve University School of MedicineAbstract Background The Lung is the major focus of therapeutic approaches for the inherited disorder cystic fibrosis (CF) as without treatment lung disease is life-limiting. However, the initiating events that predispose the CF lung to cycles of infection, inflammation and resultant tissue damage are still unclear. Inflammation may occur in the CF lung prior to birth in human and several large animal models suggesting an in utero origin for the disease and encouraging further studies prior to birth. Methods Here we used the sheep model of CF (CFTR −/− ) and age-matched wild-type (WT) sheep of the same breed to investigate the single cell transcriptomes of proximal and distal lung tissue at 80 days and 120 days of gestation and at term (147 days). Single cell RNA-seq was performed on tissues from 4 to 7 animals of each genotype (WT and CFTR −/− ) at each time point. Results At term, FOXJ1-expressing ciliated cells are overrepresented in both lung regions from CFTR −/− lambs, while secretory epithelial and basal cells are underrepresented in proximal lung, as are T cells and monocytes in distal lung. The imbalance in ciliated and basal cells was confirmed by immunohistochemistry. At 120 days of gestation, lymphoid cells are slightly more abundant in proximal and distal lung from CFTR −/− animals compared to WT, consistent with the transient CF-associated inflammatory response in utero. At 80 days of gestation, T and B cells are underrepresented in both lung regions. Conclusions The differences in epithelial cell abundance observed in the CFTR −/− lambs at term may reflect sequelae from the loss of CFTR on lung development and differentiation in utero. These findings provide novel insights into the cellular mechanisms of pathology and may be relevant to the design of new therapeutic approaches for CF lung disease.https://doi.org/10.1186/s10020-025-01266-7Cystic fibrosis (CF)In uteroLung developmentScRNA-seqCiliated cells
spellingShingle Shih-Hsing Leir
Svyatoslav Tkachenko
Alekh Paranjapye
Arnaud J. Van Wettere
Jenny L. Kerschner
Iuri Viotti Perisse
Cheyenne M. Marriott
Tayler Patrick
Ying Liu
Kenneth L. White
Irina A. Polejaeva
Ann Harris
Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
Molecular Medicine
Cystic fibrosis (CF)
In utero
Lung development
ScRNA-seq
Ciliated cells
title Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
title_full Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
title_fullStr Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
title_full_unstemmed Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
title_short Cellular imbalance in proximal and distal lung of CFTR −/− sheep in utero and at birth
title_sort cellular imbalance in proximal and distal lung of cftr sheep in utero and at birth
topic Cystic fibrosis (CF)
In utero
Lung development
ScRNA-seq
Ciliated cells
url https://doi.org/10.1186/s10020-025-01266-7
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