Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells
Systemic glutathione deficiency, inflammation, and oxidative stress are hallmarks of cystic fibrosis (CF), an inherited disease that causes persistent lung infections and severe damage to the respiratory system and many of the body organs. Improvements to current antioxidant therapeutic strategies a...
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| Format: | Article |
| Language: | English |
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MDPI AG
2020-11-01
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| Series: | Antioxidants |
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| Online Access: | https://www.mdpi.com/2076-3921/9/12/1204 |
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| author | Chris K. Hewson Alexander Capraro Sharon L. Wong Elvis Pandzic Ling Zhong Bentotage S. M. Fernando Nikhil T. Awatade Gene Hart-Smith Renee M. Whan Shane R. Thomas Adam Jaffe Wallace J. Bridge Shafagh A. Waters |
| author_facet | Chris K. Hewson Alexander Capraro Sharon L. Wong Elvis Pandzic Ling Zhong Bentotage S. M. Fernando Nikhil T. Awatade Gene Hart-Smith Renee M. Whan Shane R. Thomas Adam Jaffe Wallace J. Bridge Shafagh A. Waters |
| author_sort | Chris K. Hewson |
| collection | DOAJ |
| description | Systemic glutathione deficiency, inflammation, and oxidative stress are hallmarks of cystic fibrosis (CF), an inherited disease that causes persistent lung infections and severe damage to the respiratory system and many of the body organs. Improvements to current antioxidant therapeutic strategies are needed. The dietary supplement, γ-glutamylcysteine (GGC), which is the immediate precursor to glutathione, rapidly boosts cellular glutathione levels following a single dose in healthy individuals. Efficacy of GGC against oxidative stress induced by <i>Pseudomonas aeruginosa</i>, which is a common and chronic pathogen infecting lungs of CF patients, remains unassessed. Primary mucocilliary differentiated airway (bronchial and/or nasal) epithelial cells were created from four individuals with CF. Airway oxidative stress and inflammation was induced by <i>P. aeruginosa</i> lipopolysaccharide (LPS). Parameters including global proteomics alterations, cell redox state (glutathione, oxidative stress), pro-inflammatory mediators (IL-8, IDO-1), and cellular health (membrane integrity, stress granule formation, cell metabolic viability) were assayed under six experimental conditions: (1) Mock, (2) LPS-challenged (3) therapeutic, (4) prophylactic (5) therapeutic and prophylactic and (6) GGC alone. Proteomic analysis identified perturbation of several pathways related to cellular respiration and stress responses upon LPS challenge. Most of these were resolved when cells were treated with GGC. While GGC did not resolve LPS-induced IL-8 and IDO-1 activity, it effectively attenuated LPS-induced oxidative stress and stress granule formation, while significantly increasing total intracellular glutathione levels, metabolic viability and improving epithelial cell barrier integrity. Both therapeutic and prophylactic treatments were successful. Together, these findings indicate that GGC has therapeutic potential for treatment and prevention of oxidative stress-related damage to airways in cystic fibrosis. |
| first_indexed | 2024-03-10T14:26:38Z |
| format | Article |
| id | doaj.art-ded626cf578c43c38dbe8569a71c6601 |
| institution | Directory Open Access Journal |
| issn | 2076-3921 |
| language | English |
| last_indexed | 2024-03-10T14:26:38Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Antioxidants |
| spelling | doaj.art-ded626cf578c43c38dbe8569a71c66012023-11-20T22:58:40ZengMDPI AGAntioxidants2076-39212020-11-01912120410.3390/antiox9121204Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial CellsChris K. Hewson0Alexander Capraro1Sharon L. Wong2Elvis Pandzic3Ling Zhong4Bentotage S. M. Fernando5Nikhil T. Awatade6Gene Hart-Smith7Renee M. Whan8Shane R. Thomas9Adam Jaffe10Wallace J. Bridge11Shafagh A. Waters12School of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaBiomedical Imaging Facility, University of New South Wales, Sydney, NSW 2052, AustraliaBioanalytical Mass Spectrometry Facility, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Biotechnology and Biomolecular Sciences, Faculty of Science, University of New South Wales, Sydney, NSW 2052, AustraliaBiomedical Imaging Facility, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Medical Sciences, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Biotechnology and Biomolecular Sciences, Faculty of Science, University of New South Wales, Sydney, NSW 2052, AustraliaSchool of Women’s and Children’s Health, Faculty of Medicine, University of New South Wales, Sydney, NSW 2052, AustraliaSystemic glutathione deficiency, inflammation, and oxidative stress are hallmarks of cystic fibrosis (CF), an inherited disease that causes persistent lung infections and severe damage to the respiratory system and many of the body organs. Improvements to current antioxidant therapeutic strategies are needed. The dietary supplement, γ-glutamylcysteine (GGC), which is the immediate precursor to glutathione, rapidly boosts cellular glutathione levels following a single dose in healthy individuals. Efficacy of GGC against oxidative stress induced by <i>Pseudomonas aeruginosa</i>, which is a common and chronic pathogen infecting lungs of CF patients, remains unassessed. Primary mucocilliary differentiated airway (bronchial and/or nasal) epithelial cells were created from four individuals with CF. Airway oxidative stress and inflammation was induced by <i>P. aeruginosa</i> lipopolysaccharide (LPS). Parameters including global proteomics alterations, cell redox state (glutathione, oxidative stress), pro-inflammatory mediators (IL-8, IDO-1), and cellular health (membrane integrity, stress granule formation, cell metabolic viability) were assayed under six experimental conditions: (1) Mock, (2) LPS-challenged (3) therapeutic, (4) prophylactic (5) therapeutic and prophylactic and (6) GGC alone. Proteomic analysis identified perturbation of several pathways related to cellular respiration and stress responses upon LPS challenge. Most of these were resolved when cells were treated with GGC. While GGC did not resolve LPS-induced IL-8 and IDO-1 activity, it effectively attenuated LPS-induced oxidative stress and stress granule formation, while significantly increasing total intracellular glutathione levels, metabolic viability and improving epithelial cell barrier integrity. Both therapeutic and prophylactic treatments were successful. Together, these findings indicate that GGC has therapeutic potential for treatment and prevention of oxidative stress-related damage to airways in cystic fibrosis.https://www.mdpi.com/2076-3921/9/12/1204cystic fibrosisglutathioneredoxantioxidantoxidative stress<i>Pseudomonas aeruginosa</i> |
| spellingShingle | Chris K. Hewson Alexander Capraro Sharon L. Wong Elvis Pandzic Ling Zhong Bentotage S. M. Fernando Nikhil T. Awatade Gene Hart-Smith Renee M. Whan Shane R. Thomas Adam Jaffe Wallace J. Bridge Shafagh A. Waters Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells Antioxidants cystic fibrosis glutathione redox antioxidant oxidative stress <i>Pseudomonas aeruginosa</i> |
| title | Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells |
| title_full | Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells |
| title_fullStr | Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells |
| title_full_unstemmed | Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells |
| title_short | Novel Antioxidant Therapy with the Immediate Precursor to Glutathione, γ-Glutamylcysteine (GGC), Ameliorates LPS-Induced Cellular Stress in In Vitro 3D-Differentiated Airway Model from Primary Cystic Fibrosis Human Bronchial Cells |
| title_sort | novel antioxidant therapy with the immediate precursor to glutathione γ glutamylcysteine ggc ameliorates lps induced cellular stress in in vitro 3d differentiated airway model from primary cystic fibrosis human bronchial cells |
| topic | cystic fibrosis glutathione redox antioxidant oxidative stress <i>Pseudomonas aeruginosa</i> |
| url | https://www.mdpi.com/2076-3921/9/12/1204 |
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