2045 The role of TGFβ in driving early cystic fibrosis lung disease

OBJECTIVES/SPECIFIC AIMS: Transforming growth factor-beta (TGFβ) is a genetic modifier of cystic fibrosis (CF) lung disease. TGFβ’s pulmonary levels in young CF patients and its mechanism of action in CF are unknown. We examined TGFβ levels in children with CF and investigated responses of human airway epithelial cells (AECs) and mice to TGFβ. METHODS/STUDY POPULATION: TGFβ levels in bronchoalveolar lavage fluid from CF patients (n=15) and non-CF control patients (n=21)<6 years old were determined by ELISA. CF mice and non-CF mice were intratracheally treated with an adenoviral TGFβ1 vector or PBS; lungs were collected for analysis at day 7. Human CF and non-CF AECs were treated with TGFβ or PBS for 24 hours then collected for analysis. RESULTS/ANTICIPATED RESULTS: Young CF patients had higher bronchoalveolar lavage fluid TGFβ than non-CF controls (p=0.03). Mouse lungs exposed to TGFβ demonstrated inflammation, goblet cell hyperplasia, and decreased CFTR expression. CF mice had greater TGFβ-induced lung mechanics abnormalities than controls; both CF human AECs and CF mice showed higher TGFβ induced MAPK and PI3K signaling compared with controls. DISCUSSION/SIGNIFICANCE OF IMPACT: For the first time, we show increased TGFβ levels very early in CF. TGFβ drives CF lung abnormalities in mouse and human models; CF models are more sensitive to TGFβ’s effects. Understanding the role of TGFβ in promoting CF lung disease is critical to developing patient specific treatments.

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Transforming growth factor-β1in bronchoalveolar lavage fluid from children with cystic fibrosis

Pediatric Pulmonology ◽

10.1002/ppul.21079 ◽

2009 ◽

Vol 44 (11) ◽

pp. 1057-1064 ◽

Cited By ~ 33

Author(s):

William T. Harris ◽

Marianne S. Muhlebach ◽

Robert A. Oster ◽

Michael R. Knowles ◽

Terry L. Noah

Keyword(s):

Cystic Fibrosis ◽

Growth Factor ◽

Bronchoalveolar Lavage ◽

Bronchoalveolar Lavage Fluid ◽

Transforming Growth Factor ◽

Lavage Fluid

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Age and sex dimorphisms contribute to the severity of bleomycin-induced lung injury and fibrosis

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00122.2011 ◽

2011 ◽

Vol 301 (4) ◽

pp. L510-L518 ◽

Cited By ~ 69

Author(s):

Elizabeth F. Redente ◽

Kristen M. Jacobsen ◽

Joshua J. Solomon ◽

Abigail R. Lara ◽

Sarah Faubel ◽

...

Keyword(s):

Pulmonary Fibrosis ◽

Bronchoalveolar Lavage ◽

Lung Disease ◽

Bronchoalveolar Lavage Fluid ◽

Transforming Growth Factor ◽

Young Male ◽

Lavage Fluid ◽

Male Mice ◽

Female Mice ◽

Age And Sex

Fibrotic interstitial pneumonias are more prevalent in males of advancing age, although little is known about the underlying mechanisms. To evaluate the contributions of age and sex to the development of pulmonary fibrosis, we intratracheally instilled young (8–12 wk) and aged (52–54 wk) male and female mice with bleomycin and assessed the development and severity of fibrotic lung disease by measurements of lung collagen levels, static compliance, leukocyte infiltration, and stereological quantification of fibrotic areas in histological sections. We also quantified proinflammatory and profibrotic chemokine and cytokine levels in the bronchoalveolar lavage fluid. Aged male mice developed more severe lung disease, indicated by increased mortality, increased collagen deposition, and neutrophilic alveolitis compared with aged female mice or young mice of either sex. Aged male mice also exhibited increased levels of transforming growth factor-β, IL-17A, and CXCL1 in their bronchoalveolar lavage fluid. Young male mice developed a more fibrotic disease after bleomycin instillation compared with female mice, regardless of age. There was no difference in fibrosis between young and aged female mice. Taken together, these findings suggest that the variables of advanced age and male sex contribute to the severity of pulmonary fibrosis in this model. Our findings also emphasize the importance of stratifying experimental groups on the basis of age and sex in experimental and epidemiological studies of this nature.

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Increase in the concentration of transforming growth factor beta-1 in bronchoalveolar lavage fluid before development of chronic lung disease of prematurity

The Journal of Pediatrics ◽

10.1016/s0022-3476(96)70355-4 ◽

1996 ◽

Vol 128 (4) ◽

pp. 464-469 ◽

Cited By ~ 130

Author(s):

S. Kotecha ◽

A. Wangoo ◽

M. Silverman ◽

R.J. Shaw

Keyword(s):

Growth Factor ◽

Bronchoalveolar Lavage ◽

Lung Disease ◽

Chronic Lung Disease ◽

Transforming Growth Factor Beta ◽

Bronchoalveolar Lavage Fluid ◽

Transforming Growth Factor ◽

Lavage Fluid ◽

Growth Factor Beta

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Novel Neutrophil-Derived Proteins in Bronchoalveolar Lavage Fluid Indicate an Exaggerated Inflammatory Response in Pediatric Cystic Fibrosis Patients

Clinical Chemistry ◽

10.1373/clinchem.2007.087650 ◽

2007 ◽

Vol 53 (10) ◽

pp. 1782-1791 ◽

Cited By ~ 32

Author(s):

Brendan J McMorran ◽

Severine A Ouvry Patat ◽

John B Carlin ◽

Keith Grimwood ◽

Alun Jones ◽

...

Keyword(s):

Cystic Fibrosis ◽

Inflammatory Response ◽

Bronchoalveolar Lavage ◽

Lung Disease ◽

Airway Inflammation ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Respiratory Pathogens ◽

Tissue Destruction ◽

Tof Ms

Abstract Background: Airway inflammation in cystic fibrosis (CF) is exaggerated and characterized by neutrophil-mediated tissue destruction, but its genesis and mechanisms remain poorly understood. To further define the pulmonary inflammatory response, we conducted a proteome-based screen of bronchoalveolar lavage fluid (BALF) collected from young children with and without CF experiencing endobronchial infection. Methods: We collected BALF samples from 45 children younger than 5 years and grouped them according to the presence of respiratory pathogens: ≥1 × 105 colony-forming units (CFU)/mL BALF (18 and 12 samples with and without CF, respectively) and <1 × 105 CFU/mL (23 and 15 samples). BALF proteins were analyzed with SELDI-TOF mass spectrometry (MS) and H4 ProteinChips®. Proteins were identified and characterized using trypsin digestion, tandem MS, Fourier transform ion cyclotron resonance MS, immunoblotting, and ELISA. Results: The SELDI-TOF MS BALF profiles contained 53 unique, reliably detected proteins. Peak intensities of 24 proteins differed significantly between the CF and non-CF samples. They included the neutrophil proteins, α-defensin 1 and 2, S100A8, S100A9, and S100A12, as well as novel forms of S100A8 and S100A12 with equivalent C-terminal deletions. Peak intensities of these neutrophil proteins and immunoreactive concentrations of selected examples were significantly higher in CF than non-CF samples. Conclusions: Small neutrophil-derived BALF proteins, including novel C-terminal truncated forms of S100A proteins, are easily detected with SELDI-TOF MS. Concentrations of these molecules are abnormally high in early CF lung disease. The data provide new insights into CF lung disease and identify novel proteins strongly associated with CF airway inflammation.

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Metabolomic biomarkers predictive of early structural lung disease in cystic fibrosis

European Respiratory Journal ◽

10.1183/13993003.00524-2016 ◽

2016 ◽

Vol 48 (6) ◽

pp. 1612-1621 ◽

Cited By ~ 34

Author(s):

Charles R. Esther ◽

Lidija Turkovic ◽

Tim Rosenow ◽

Marianne S. Muhlebach ◽

Richard C. Boucher ◽

...

Keyword(s):

Cystic Fibrosis ◽

Bronchoalveolar Lavage ◽

Lung Disease ◽

Bronchoalveolar Lavage Fluid ◽

Lavage Fluid ◽

Small Peptides ◽

Neutrophilic Inflammation ◽

Cellular Energy ◽

Cystic Fibrosis Lung Disease ◽

And Oxidative Stress

Neutrophilic airway inflammation plays a role in early structural lung disease in cystic fibrosis, but the mechanisms underlying this pathway are incompletely understood.Metabolites associated with neutrophilic inflammation were identified by discovery metabolomics on bronchoalveolar lavage fluid supernatant from 20 preschool children (2.9±1.3 years) with cystic fibrosis. Targeted mass-spectrometric detection of relevant metabolites was then applied to 34 children (3.5±1.5 years) enrolled in the Australian Respiratory Early Surveillance Team for Cystic Fibrosis (AREST CF) who underwent chest computed tomography and bronchoalveolar lavage from two separate lobes during 42 visits. Relationships between metabolites and localised structural lung disease were assessed using multivariate analyses.Discovery metabolomics identified 93 metabolites associated with neutrophilic inflammation, including pathways involved in metabolism of adenyl purines, amino acids and small peptides, cellular energy and lipids. In targeted mass spectrometry, products of adenosine metabolism, protein catabolism and oxidative stress were associated with structural lung disease and predicted future bronchiectasis, and activities of enzymes associated with adenosine metabolism were elevated in the samples with early disease.Metabolomics analyses revealed metabolites and pathways altered with neutrophilic inflammation and destructive lung disease. These pathways can serve as biomarkers and potential therapeutic targets for early cystic fibrosis lung disease.

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