Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response
Background Human nasal epithelial (HNE) cells can be sampled noninvasively and cultured to provide a model of the airway epithelium that reflects cystic fibrosis (CF) pathophysiology. We hypothesised that in vitro measures of HNE cell physiology would correlate directly with in vivo measures of lung...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
European Respiratory Society
2022-12-01
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| Series: | ERJ Open Research |
| Online Access: | http://openres.ersjournals.com/content/8/4/00382-2022.full |
| _version_ | 1827931433110339584 |
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| author | Timothy E. Corcoran Carol A. Bertrand Michael M. Myerburg Daniel J. Weiner Sheila A. Frizzell Anna Li Brittani Agostini Robert S. Parker Monica E. Shapiro Ashok Muthukrishnan Nicholas D. Hages Brian P. Mulhern Joseph M. Pilewski |
| author_facet | Timothy E. Corcoran Carol A. Bertrand Michael M. Myerburg Daniel J. Weiner Sheila A. Frizzell Anna Li Brittani Agostini Robert S. Parker Monica E. Shapiro Ashok Muthukrishnan Nicholas D. Hages Brian P. Mulhern Joseph M. Pilewski |
| author_sort | Timothy E. Corcoran |
| collection | DOAJ |
| description | Background
Human nasal epithelial (HNE) cells can be sampled noninvasively and cultured to provide a model of the airway epithelium that reflects cystic fibrosis (CF) pathophysiology. We hypothesised that in vitro measures of HNE cell physiology would correlate directly with in vivo measures of lung physiology and therapeutic response, providing a framework for using HNE cells for therapeutic development and precision medicine.
Methods
We sampled nasal cells from participants with CF (CF group, n=26), healthy controls (HC group, n=14) and single CF transmembrane conductance regulator (CFTR) mutation carrier parents of the CF group (CR group, n=16). Participants underwent lung physiology and sweat chloride testing, and nuclear imaging-based measurement of mucociliary clearance (MCC) and small-molecule absorption (ABS). CF participants completed a second imaging day that included hypertonic saline (HS) inhalation to assess therapeutic response in terms of MCC. HNE measurements included Ussing chamber electrophysiology, small-molecule and liquid absorption rates, and particle diffusion rates through the HNE airway surface liquid (ASL) measured using fluorescence recovery after photobleaching (FRAP).
Results
Long FRAP diffusion times were associated with increased MCC response to HS in CF. This implies a strong relationship between inherent factors affecting ASL mucin concentration and therapeutic response to a hydrating therapy. MCC decreased with age in the CR group, which had a larger range of ages than the other two groups. Likely this indicates a general age-related effect that may be accentuated in this group. Measures of lung ABS correlated with sweat chloride in both the HC and CF groups, indicating that CFTR function drives this measure of paracellular small-molecule probe absorption.
Conclusions
Our results demonstrate the utility of HNE cultures for assessing therapeutic response for hydrating therapies. In vitro measurements of FRAP were particularly useful for predicting response and for characterising important properties of ASL mucus that were ultimately reflected in lung physiology. |
| first_indexed | 2024-03-13T06:52:35Z |
| format | Article |
| id | doaj.art-4cfa57bff8bf44f994aa6ea60489b03d |
| institution | Directory Open Access Journal |
| issn | 2312-0541 |
| language | English |
| last_indexed | 2024-03-13T06:52:35Z |
| publishDate | 2022-12-01 |
| publisher | European Respiratory Society |
| record_format | Article |
| series | ERJ Open Research |
| spelling | doaj.art-4cfa57bff8bf44f994aa6ea60489b03d2023-06-07T13:30:37ZengEuropean Respiratory SocietyERJ Open Research2312-05412022-12-018410.1183/23120541.00382-202200382-2022Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic responseTimothy E. Corcoran0Carol A. Bertrand1Michael M. Myerburg2Daniel J. Weiner3Sheila A. Frizzell4Anna Li5Brittani Agostini6Robert S. Parker7Monica E. Shapiro8Ashok Muthukrishnan9Nicholas D. Hages10Brian P. Mulhern11Joseph M. Pilewski12 Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA Department of Pediatrics, University of Pittsburgh, Pittsburgh, PA, USA Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA Division of Pediatric Pulmonology, University of Pittsburgh, Pittsburgh, PA, USA Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA School of Medicine, University of Pittsburgh, Pittsburgh, PA, USA Division of Pediatric Pulmonology, University of Pittsburgh, Pittsburgh, PA, USA Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA Department of Radiology, University of Pittsburgh, Pittsburgh, PA, USA Department of Chemical and Petroleum Engineering, University of Pittsburgh, Pittsburgh, PA, USA Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA Division of Pulmonary, Allergy, and Critical Care Medicine, University of Pittsburgh, Pittsburgh, PA, USA Background Human nasal epithelial (HNE) cells can be sampled noninvasively and cultured to provide a model of the airway epithelium that reflects cystic fibrosis (CF) pathophysiology. We hypothesised that in vitro measures of HNE cell physiology would correlate directly with in vivo measures of lung physiology and therapeutic response, providing a framework for using HNE cells for therapeutic development and precision medicine. Methods We sampled nasal cells from participants with CF (CF group, n=26), healthy controls (HC group, n=14) and single CF transmembrane conductance regulator (CFTR) mutation carrier parents of the CF group (CR group, n=16). Participants underwent lung physiology and sweat chloride testing, and nuclear imaging-based measurement of mucociliary clearance (MCC) and small-molecule absorption (ABS). CF participants completed a second imaging day that included hypertonic saline (HS) inhalation to assess therapeutic response in terms of MCC. HNE measurements included Ussing chamber electrophysiology, small-molecule and liquid absorption rates, and particle diffusion rates through the HNE airway surface liquid (ASL) measured using fluorescence recovery after photobleaching (FRAP). Results Long FRAP diffusion times were associated with increased MCC response to HS in CF. This implies a strong relationship between inherent factors affecting ASL mucin concentration and therapeutic response to a hydrating therapy. MCC decreased with age in the CR group, which had a larger range of ages than the other two groups. Likely this indicates a general age-related effect that may be accentuated in this group. Measures of lung ABS correlated with sweat chloride in both the HC and CF groups, indicating that CFTR function drives this measure of paracellular small-molecule probe absorption. Conclusions Our results demonstrate the utility of HNE cultures for assessing therapeutic response for hydrating therapies. In vitro measurements of FRAP were particularly useful for predicting response and for characterising important properties of ASL mucus that were ultimately reflected in lung physiology.http://openres.ersjournals.com/content/8/4/00382-2022.full |
| spellingShingle | Timothy E. Corcoran Carol A. Bertrand Michael M. Myerburg Daniel J. Weiner Sheila A. Frizzell Anna Li Brittani Agostini Robert S. Parker Monica E. Shapiro Ashok Muthukrishnan Nicholas D. Hages Brian P. Mulhern Joseph M. Pilewski Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response ERJ Open Research |
| title | Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| title_full | Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| title_fullStr | Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| title_full_unstemmed | Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| title_short | Nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| title_sort | nasal epithelial cell culture fluorescence recovery after photobleaching predicts cystic fibrosis therapeutic response |
| url | http://openres.ersjournals.com/content/8/4/00382-2022.full |
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