MECHANISMS OF TGFβ ACTIVATION IN LUNG FIBROSIS

Abstract

<p>This PhD thesis focuses on the mechanical activation of TGFβ in the context of pulmonary fibrosis. Mechanical TGFβ activation occurs by physical force of breathing and signals to the nucleus via phospho‐Smad2. This activation occurs in presence of strong pan‐serine protease and matrix metalloproteinase inhibition. The augmented expression of latent TGFβ in lung tissue also lead to TGFβ activity following tissue stretch. Tissue biopsies from pulmonary fibrosis patients exhibited the same mechanical TGFβ activation and subsequent accumulation of phospho‐Smad2 as was seen in animal models. In rodent models and human control tissue, TGFβ was not released in detectable quantities, nor was there any significant upregulation of phospho‐Smad2. These data show that mechanical TGFβ activation is a relevant and limited to the context of a fibrotic disease process. Non‐invasive investigation of lung fibrosis was evaluated for correlation to classical assessments. We found that non‐invasive lung function parameters measured by a rodent ventilator, and small animal CT imaging correlated significantly with histomorphometic Ashcroft scoring. Exercise testing and quantification of the maximal oxygen consumption rate was a valuable indicator of overall rodent lung health but did not correlated significantly with Ashcroft scoring. Non‐invasive investigation tools evaluated here represent important advances in the quality of interpretation of preclinical lung fibrosis trials. Finally, collagen turnover was investigated by measurement of pyridinolines and serum collagen metabolic peptides. A novel method was developed and tested to detect pyridinolines in facile procedure. We found that deoxypyridinolines, but not pyridinolines, were significantly increased in the serum of lung fibrosis patients with respect to healthy controls. Furthermore, collagen type 1 telopeptide, a collagen breakdown product, was significantly increased in lung fibrosis patient serum. These data intriguingly indicate that under stable lung fibrosis conditions, more collagen appears to be breaking down into the serum then is synthesized.</p>