Fibroblasts as matrix modulating cells in asthma and COPD

Chronic lung diseases such as asthma and chronic obstructive pulmonary disease (COPD) are two lung diseases that are continuously increasing worldwide. Despite extensively research to find curative treatment, no so such therapy exists today. New hypothesis suggests that an aberrant chronic wound healing process takes place which involves both the classical inflammation and more chronic changes of the structural environment where the mesenchymal cell phenotypes such as the fibroblasts play a key role. This phenomenon is defined as airway remodeling and is characterized by a deposition of extracellular matrix (ECM) molecules such as collagens and proteoglycans. Importantly, hitherto available studies have only studied fibroblast from central airways from humans where the role of the fibroblast might be different compared to the distal lung. The aim of the present thesis was to study the different fibroblast phenotypes and their biological role from central and distal localizations from human lung in controls, rhinitis, asthmatics and CODP subjects. Another aim was to investigate the possible origin (circulating progenitor cells) of these cells and thereby highlight the heterogeneity of fibroblast phenotypes resident within the human lung. This investigation demonstrated the novel finding that different fibroblast phenotypes are present in human adult lung. Moreover, they are different in asthma and COPD which suggest disease related fibroblasts that could be of fundamental importance in lung disease

Enhanced ROCK1 dependent contractility in fibroblast from chronic obstructive pulmonary disease patients

Background: During wound healing processes fibroblasts account for wound closure by adopting a contractile phenotype. One disease manifestation of COPD is emphysema which is characterized by destruction of alveolar walls and our hypothesis is that fibroblasts in the COPD lungs differentiate into a more contractile phenotype as a response to the deteriorating environment. Methods: Bronchial (central) and parenchymal (distal) fibroblasts were isolated from lung explants from COPD patients (n = 9) (GOLD stage IV) and from biopsies from control subjects and from donor lungs (n = 12). Tissue-derived fibroblasts were assessed for expression of proteins involved in fibroblast contraction by western blotting whereas contraction capacity was measured in three-dimensional collagen gels. Results: The basal expression of rho-associated coiled-coil protein kinase 1 (ROCK1) was increased in both centrally and distally derived fibroblasts from COPD patients compared to fibroblasts from control subjects (p < 0.001) and (p < 0.01), respectively. Distally derived fibroblasts from COPD patients had increased contractile capacity compared to control fibroblasts (p < 0.01). The contraction was dependent on ROCK1 activity as the ROCK inhibitor Y27632 dose-dependently blocked contraction in fibroblasts from COPD patients. ROCK1-positive fibroblasts were also identified by immunohistochemistry in the alveolar parenchyma in lung tissue sections from COPD patients. Conclusions: Distally derived fibroblasts from COPD patients have an enhanced contractile phenotype that is dependent on ROCK1 activity. This feature may be of importance for the elastic dynamics of small airways and the parenchyma in late stages of COPD

Leukotriene receptors are differently expressed in fibroblast from peripheral versus central airways in asthmatics and healthy controls.

Leukotrienes are involved in airway inflammation, and are believed to stimulate airway remodeling in asthma. The aim of the project was to investigate the expression of leukotriene receptors in peripheral and central airway fibroblasts. Peripheral and central airway fibroblasts, from asthmatics and healthy controls, were investigated for the amount of cysteinyl-leukotriene receptors (CysLT(1) and CysLT(2)), leukotriene B(4) receptors (BLT(1) and BLT(2)), IL-13 receptor-α(1) (IL-13Rα(1)) and the IL-4 receptor (IL-4R). The mRNA expression of CysLT(1) in fibroblasts from peripheral airways was higher compared to central airways. There was no difference in CysLT(2) between peripheral and central airways. On the contrary, BLT(1) and BLT(2) were lower in fibroblasts from peripheral airways compared to central. The expression of CysLT(1) was higher than CysLT(2) in fibroblasts from peripheral airways, and the expression of BLT(1) was higher than BLT(2) in both peripheral and central airways. Both BLT(1) and BLT(2) were higher in asthmatics compared to healthy controls, while CysLT(1) and CysLT(2) did not differ. The expression of IL-13Rα(1) was higher in asthmatics compared to controls, and correlated to the BLTs. All fibroblasts stained for the different receptor proteins. Leukotriene receptors are differently expressed in fibroblasts from peripheral compared to central airways, which may explain a suggested cysteinyl-leukotriene driven remodeling mainly in the peripheral airways

Tissue fibrocytes in patients with mild asthma: A possible link to thickness of reticular basement membrane?

Abstract Background Myofibroblasts, proposed as being derived from circulating fibrocytes, are considered to be important cells in thickening of the basement membrane in patients with asthma. We have studied the correlation of tissue fibrocyte levels to basement membrane thickness and the presence of fibrocytes in bronchoalveolar lavage fluid (BALF) in steroid-naive patients with mild asthma and controls. Methods Patients with mild asthma (n = 9) were recruited and divided into two categories based on whether or not fibroblast-like cells could be established from BALF. Non-asthmatic healthy subjects (n = 5) were used as controls. Colocalization of the fibrocyte markers CD34, CD45RO, procollagen I, and α-smooth muscle actin (α-SMA) were identified in bronchial biopsies from patients and controls by confocal microscopy. Kruskall-Wallis method was used to calculate statistical significance and Spearman coefficient of rank correlation was used to assess the degree of association. Results In patients with BALF fibroblasts, a 14-fold increase of tissue cells expressing CD34/CD45RO/α-SMA and a 16-fold increase of tissue cells expressing CD34/procollagen I was observed when compared to controls (p Conclusion These findings indicate a correlation between recruited fibrocytes in tissue and thickness of basement membrane. Fibroblast progenitor cells may therefore be important in airway remodeling in steroid-naive patients with mild asthma.</p

Altered matrix production in the distal airways of individuals with asthma.

BACKGROUND AND AIMS: Although increasing evidence suggests involvement of the distal airway in all stages of asthma, it is not known whether structural changes (defined as airway remodelling) occur in the distal airways of subjects with mild asthma and those with atopy. The aim of this study was to compare control subjects and those with mild asthma in relation to fibroblast phenotypes and remodelling in central and distal airways. METHODS: Distal and central fibroblasts from controls (n=12) and patients with mild asthma (n=11) were cultured and incubated for 24 h with 0.4% serum, or stimulated with transforming growth factor beta1 (TGFbeta1). [(35)S]Sulfate-labelled proteoglycans in culture medium were analysed by ion exchange chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Proliferation was measured with crystal violet, and exhaled nitric oxide was measured by the fractional nitric oxide technique. RESULTS: Vesican production from distal fibroblasts was significantly elevated in patients with asthma compared with controls (p<0.001), and the percentage collagen-positive area in distal asthma tissue was also enhanced compared with controls (p<0.01). In addition, distal asthma fibroblasts had reduced proliferation capacity compared with those of controls (by 24%; p<0.01). Furthermore, the alveolar nitric oxide concentration was correlated to distal biglycan and perlecan production of subjects with asthma (r=-0.857, p<0.05 and r=-0.750, p<0.05 respectively) CONCLUSION: It is shown that centrally and distally derived fibroblasts differ in their proteoglycan production and proliferation between central and distal tissue, and in those with asthma compared with controls. It is also demonstrated that remodelling is present in distal lung of subjects with mild asthma. This may be of importance in airway remodelling and asthma progression

Tissue fibrocytes in patients with mild asthma: A possible link to thickness of reticular basement membrane?

Background: Myofibroblasts, proposed as being derived from circulating fibrocytes, are considered to be important cells in thickening of the basement membrane in patients with asthma. We have studied the correlation of tissue fibrocyte levels to basement membrane thickness and the presence of fibrocytes in bronchoalveolar lavage fluid (BALF) in steroid-naive patients with mild asthma and controls. Methods: Patients with mild asthma (n=9) were recruited and divided into two categories based on whether or not fibroblast-like cells could be established from BALF. Non-asthmatic healthy subjects (n=5) were used as controls. Colocalization of the fibrocyte markers CD34, CD45RO, procollagen I, and alpha-smooth muscle actin (alpha-SMA) were identified in bronchial biopsies from patients and controls by confocal microscopy. Kruskall-Wallis method was used to calculate statistical significance and Spearman coefficient of rank correlation was used to assess the degree of association. Results: In patients with BALF fibroblasts, a 14-fold increase of tissue cells expressing CD34/CD45RO/alpha-SMA and a 16-fold increase of tissue cells expressing CD34/procollagen I was observed when compared to controls (p<0.05). In contrast, patients without BALF fibroblasts displayed a 2-fold increase when compared to controls (p<0.05). Fibrocytes were localized close to the basement membrane which was significantly thicker in patients with BALF fibroblasts when compared to the other two groups of subjects. Furthermore, basement membrane thickness could be correlated to the number of fibrocytes in tissue (r=0.711). Fibroblasts-like cells were cultured from BALF where 17.6% of these cells expressed CD34, CD45RO and alpha-SMA. Conclusion: These findings indicate a correlation between recruited fibrocytes in tissue and thickness of basement membrane. Fibroblast progenitor cells may therefore be important in airway remodeling in steroid-naive patients with mild asthma

Efficient induction of functional neurons from adult human fibroblasts.

Cellular reprogramming is a rapidly developing technology by which somatic cells are turned into pluripotent stem cells or other somatic cell types through expression of specific combinations of genes. This allows for the generation of patient-specific cell lines that can serve as tools for understanding disease pathogenesis, for drug screens and, potentially, for cell replacement therapies. Several cellular models of neurological disorders based on induced pluripotent cells (iPS cells) have been developed, and iPS-derived neurons are being explored as candidates for transplantation. Recent findings show that neurons can also be induced directly from embryonic and postnatal somatic cells by expression of defined combinations of genes. This conversion does not occur through a pluripotent stem cell stage, which eliminates the risk for tumor formation. Here, we demonstrate for the first time that functional neurons can be generated via direct conversion of fibroblasts also from adult individuals. Thus, this technology is an attractive alternative to iPS cells for generating patient- and disease-specific neurons suitable for disease modeling and autologous transplantation