Rolf Ziesche

A Preliminary Study of Long-Term Treatment with Interferon Gamma-1b and Low-Dose Prednisolone in Patients with Idiopathic Pulmonary Fibrosis

BACKGROUND AND METHODS Patients with idiopathic pulmonary fibrosis have progressive scarring of the lung and usually die within four to five years after symptoms develop. Treatment with oral glucocorticoids is often ineffective. We conducted an open, randomized trial of treatment with a combination of interferon gamma-1b, which has antifibrotic properties, and an oral glucocorticoid. We studied 18 patients with idiopathic pulmonary fibrosis who had not had responses to glucocorticoids or other immunosuppressive agents. Nine patients were treated for 12 months with oral prednisolone alone (7.5 mg daily, which could be increased to 25 to 50 mg daily), and nine with a combination of 200 microg of interferon gamma-1b (given three times per week subcutaneously) and 7.5 mg of prednisolone (given once a day). RESULTS All the patients completed the study. Lung function deteriorated in all nine patients in the group given prednisolone alone: total lung capacity decreased from a mean (+/-SD) of 66+/-8 percent of the predicted value at base line to 62+/-6 percent at 12 months. In contrast, in the group receiving interferon gamma-1b plus prednisolone, total lung capacity increased (from 70+/-6 percent of the predicted value at base line to 79+/-12 percent at 12 months, P<0.001 for the difference between the groups). In the group that received interferon gamma-1b plus prednisolone, the partial pressure of arterial oxygen at rest increased from 65+/-9 mm Hg at base line to 76+/-8 mm Hg at 12 months, whereas in the group that received prednisolone alone it decreased from 65+/-6 to 62+/-4 mm Hg (P<0.001 for the difference in the change from baseline values between the two groups); on maximal exertion, the value increased from 55+/-6 to 65+/-8 mm Hg in the group that received combined treatment and decreased from 55+/-6 mm Hg to 52+/-5 mm Hg in the group given prednisolone alone (P<0.001). The side effects of interferon gamma-1b, such as fever, chills, and muscle pain, subsided within the first 9 to 12 weeks. CONCLUSIONS In a preliminary study, 12 months of treatment with interferon gamma-1b plus prednisolone was associated with substantial improvements in the condition of patients with idiopathic pulmonary fibrosis who had had no response to glucocorticoids.

Interaction of C/EBPα and the glucocorticoid receptor in vivo and in nontransformed human cells

Belonging to the family of steroid hormones, glucocorticoids are essential for development and survival of vertebrates. The cellular response to glucocorticoids is attributed to the glucocorticoid receptor, which functions as a transcription factor. However, the majority of glucocorticoid‐modulated genes lack a DNA binding site for the glucocorticoid receptor, raising the question of which mechanism mediates the responses to glucocorticoids. It has been suggested that besides direct DNA binding of the glucocorticoid receptor, interaction with members of other transcription factor families modulates the effect of the glucocorti‐coid receptor. However, the significance of such transcription factor interaction is not clear. In cultured human mesenchymal cells and peripheral blood leukocytes of human volunteers treated with glucocorticoids, we detected the formation of a complex between the GR and the CCAAT/enhancer binding protein a.Inin vitro experiments, this interaction turned out to be responsible for the inhibitory action of glucocorticoids on lymphocytic and mesenchymal cell proliferation. Our results suggest that complex formation of the GR with C/EBPα accounts for a novel pathway of glucocorticoid action.—Rüdiger, J. J., Roth, M., Bihl, M. P., Cornelius, B. C., Johnson, M., Ziesche, R., Block, L.‐H. Interaction of C/EBPα and the glucocorticoid receptor in vivo and in nontransformed human cells. FASEB J. 16, 177–184 (2002)

Diagnostic Performance of Plasma DNA Methylation Profiles in Lung Cancer, Pulmonary Fibrosis and COPD.

Disease-specific alterations of the cell-free DNA methylation status are frequently found in serum samples and are currently considered to be suitable biomarkers. Candidate markers were identified by bisulfite conversion-based genome-wide methylation screening of lung tissue from lung cancer, fibrotic ILD, and COPD. cfDNA from 400 μl serum (n = 204) served to test the diagnostic performance of these markers. Following methylation-sensitive restriction enzyme digestion and enrichment of methylated DNA via targeted amplification (multiplexed MSRE enrichment), a total of 96 markers were addressed by highly parallel qPCR. Lung cancer was efficiently separated from non-cancer and controls with a sensitivity of 87.8%, (95%CI: 0.67–0.97) and specificity 90.2%, (95%CI: 0.65–0.98). Cancer was distinguished from ILD with a specificity of 88%, (95%CI: 0.57–1), and COPD from cancer with a specificity of 88% (95%CI: 0.64–0.97). Separation of ILD from COPD and controls was possible with a sensitivity of 63.1% (95%CI: 0.4–0.78) and a specificity of 70% (95%CI: 0.54–0.81). The results were confirmed using an independent sample set (n = 46) by use of the four top markers discovered in the study (HOXD10, PAX9, PTPRN2, and STAG3) yielding an AUC of 0.85 (95%CI: 0.72–0.95). This technique was capable of distinguishing interrelated complex pulmonary diseases suggesting that multiplexed MSRE enrichment might be useful for simple and reliable diagnosis of diverse multifactorial disease states.

Abnormal pulmonary arterial pressure limits exercise capacity in patients with COPD

ZusammenfassungZIELE: Das Vorliegen einer pulmonalen Hypertension ist häufig bei Patienten mit chronisch obstruktiver Lungenerkrankung (COPD) anzutreffen. Der mittlere pulmonalarterielle Druck (mPAP) ist in Ruhe oft nur gering erhöht, zeigt aber einen pathologischen Anstieg unter Belastung. Das Ziel dieser Studie ist es, die Leistungsfähigkeit und den pulmonalen Gasaustausch bei COPD Patienten mit und ohne pulmonalarterieller Hypertension zu untersuchen. PATIENTEN UND METHODEN: Bei 42 Patienten mit COPD Grad II-IV (28 Männer, 14 Frauen) wurden eine Bodyplethysmographie, eine symptomlimitierte Fahrradergospirometrie sowie eine Rechtsherzkatheteruntersuchung durchgeführt. RESULTATE: 32 von 42 Patienten (76%) zeigten einen erhöhten mPAP in Ruhe (PH mPAP = 26,8 ± 5,9 mmHg), bei 10 Patienten war der mPAP in Ruhe im Normbereich (NPH, mPAP = 16,8 ± 2 mmHg). Es gab keinen signifikanten Unterschied hinsichtlich der lungenfunktionellen Parameter in beiden Gruppen. Die maximale Sauerstoffaufnahme (VO2max) war signifikant niedriger in der PH Gruppe (785 ± 244 ml/min) im Vergleich zur NPH Gruppe (1052 ± 207 ml/min, p = 0,004). Es zeigte sich in der PH Gruppe eine erhöhte Totraumventilation mit signifikant erhöhtem Atemäquivalent für CO2 (VECO2 47,3 ± 10 vs 38,6 ± 3,5, p = 0,025) und signifikant höherem arterio-endtidalen CO2 Partialdruck [p(a-ET)CO2]. Der pulmonalarterielle Widerstand (PVR) in Ruhe zeigte eine negative Korrelation hinsichtlich der VO2max, VE/VCO2 und dem arterio-endtidalen CO2 Partialdruck [p(a-ET)CO2]. ZUSAMMENFASSUNG: Patienten mit COPD und erhöhter pulmonalarterieller Druckwerte in Ruhe zeigen eine Verschlechterung des pulmonalen Gasaustausches unter Belastung, eine Beeinträchtigung der maximalen Sauerstoffaufnahme und somit eine limitierte Leistungsfähigkeit.SummaryOBJECTIVE: Pulmonary hypertension (PH) is common in patients with chronic obstructive pulmonary disease (COPD). Mean pulmonary artery pressure (mPAP) is often only slightly elevated at rest but is increased by exercise. The purpose of this study was to determine whether abnormal pulmonary artery pressure impairs exercise capacity in patients with COPD. PATIENTS AND METHODS: 42 patients with moderate-to-very-severe COPD (28 men, 14 women) underwent symptom-limited incremental cardiopulmonary exercise testing and also right-heart catheterization at rest. Abnormal pulmonary artery pressure was defined as mPAP > 20 mmHg at rest. RESULTS: Resting mPAP was elevated in 32 patients (PH, mPAP = 26.8 ± 5.9 mmHg) and normal in 10 non-hypertensive (NPH) patients (NPH, mPAP = 16.8 ± 2 mmHg). There were no significant differences in lung function between the PH and NPH groups. Maximum oxygen uptake during exercise (VO2max) was significantly lower in PH (785 ± 244 ml/min) than in NPH (1052 ± 207 ml/min, P = 0.004). Dead-space ventilation (Vd/Vt) was greater in PH (P = 0.05) with higher VE/VCO2 (ratio of minute ventilation to carbon dioxide output = 47.3 ± 10 vs 38.6 ± 3.5, P = 0.025) and significantly higher arterial-end-tidal pCO2 difference [p(a-ET)CO2]. Pulmonary vascular resistance measured at rest correlated significantly with VO2max, VE/VCO2 and p(a-ET)CO2. CONCLUSIONS: In patients with COPD, abnormal pulmonary artery pressure impairs gas exchange, decreases maximum oxygen uptake during exercise and impairs exercise capacity.

Assessment of gene transcription demonstrates connection with the clinical course of idiopathic interstitial pneumonia.

Background: Mediators of preferably mesenchymal repair such as transforming growth factor β1 (TGF-β1) and mediators of polarized cellular immunity such as interleukin (IL)-13 are thought to be of key importance for progression of lung fibrosis. Nonetheless, a correlation between these mediators and the clinical development of fibrosis has not been performed thus far. Objectives: We correlated the transcription of TGF-β1, connective tissue growth factor (CTGF), IL-4, IL-13 and interferon-γ (IFN-γ) with lung function development in progressive fibrosis in idiopathic interstitial pneumonia. Methods: One hundred seventy nine sets of RT-PCR measurements were analyzed in 49 patients with usual interstitial pneumonia, nonspecific interstitial pneumonia or both. Specimens were taken by surgical and transbronchial lung biopsy. Lung function was measured at the time of biopsy and 1 year later. All patients received conventional treatment. Thirteen individuals were used as controls. Results: Transcription of TGF-β1, CTGF and IL-13 was significantly higher in pulmonary fibrosis compared to control, whereas transcription of IFN-γ and IL-4 was virtually absent in both normal and fibrotic lungs. When comparing gene transcription with development of lung function, a significant correlation was observed between the decrease in both vital capacity and total lung capacity and increased transcription levels of TGF-β1 and IL-13. A reduced pulmonary diffusion capacity correlated with increased levels of TGF-β1 and CTGF. Transcription pattern in transbronchial and surgical samples was similar. Conclusions: We found a significant correlation between gene transcription and decrease in lung function that was more pronounced for TGF-β1 than for CTGF or IL-13. Our results suggest that transcription analysis may be used in clinical assessment of pulmonary fibrosis.

Epithelial to mesenchymal transition-related proteins ZEB1, β -catenin, and β -tubulin-III in idiopathic pulmonary fibrosis

Epithelial to mesenchymal transition has been suggested as a relevant contributor to pulmonary fibrosis, but how and where this complex process is triggered in idiopathic pulmonary fibrosis is not fully understood. Beta-tubulin-III (Tubβ3), ZEB1, and β-catenin are partially under the negative control of miR-200, a family of micro-RNAs playing a major role in epithelial to mesenchymal transition, that are reduced in experimental lung fibrosis and idiopathic pulmonary fibrosis. We wonder whether in situ expression of these proteins is increased in idiopathic pulmonary fibrosis, to better understand the significance of miR-200 feedback loop and epithelial to mesenchymal transition. We investigated the immunohistochemical and immunofluorescent expression and precise location of ZEB1, Tubβ3, and β-catenin in tissue samples from 34 idiopathic pulmonary fibrosis cases and 21 controls (5 normal lungs and 16 other interstitial lung diseases). In 100% idiopathic pulmonary fibrosis samples, the three proteins were concurrently expressed in fibroblastic foci, as well in damaged epithelial cells overlying these lesions and in pericytes within neo-angiogenesis areas. These results were also confirmed by immunofluorescence assay. In controls the abnormal expression of the three proteins was absent or limited. This is the first study that relates concurrent expression of Tubβ3, ZEB1, and β-catenin to abnormal epithelial and myofibroblast differentiation in idiopathic pulmonary fibrosis, providing indirect but robust evidence of miR-200 deregulation and epithelial to mesenchymal transition activation in idiopathic pulmonary fibrosis. The abnormal expression and localization of these proteins in bronchiolar fibro-proliferative lesions are unique for idiopathic pulmonary fibrosis, and might represent a disease-specific marker in challenging lung biopsies.

Tissue repair genes: the TiRe database and its implication for skin wound healing

Wound healing is an inherent feature of any multicellular organism and recent years have brought about a huge amount of data regarding regular and abnormal tissue repair. Despite the accumulated knowledge, modulation of wound healing is still a major biomedical challenge, especially in advanced ages. In order to collect and systematically organize what we know about the key players in wound healing, we created the TiRe (Tissue Repair) database, an online collection of genes and proteins that were shown to directly affect skin wound healing. To date, TiRe contains 397 entries for four organisms: Mus musculus, Rattus norvegicus, Sus domesticus, and Homo sapiens. Analysis of the TiRe dataset of skin wound healing-associated genes showed that skin wound healing genes are (i) over-conserved among vertebrates, but are under-conserved in invertebrates; (ii) enriched in extracellular and immuno-inflammatory genes; and display (iii) high interconnectivity and connectivity to other proteins. The latter may provide potential therapeutic targets. In addition, a slower or faster skin wound healing is indicative of an aging or longevity phenotype only when assessed in advanced ages, but not in the young. In the long run, we aim for TiRe to be a one-station resource that provides researchers and clinicians with the essential data needed for a better understanding of the mechanisms of wound healing, designing new experiments, and the development of new therapeutic strategies. TiRe is freely available online at http://www.tiredb.org.

Konsensus-Empfehlungen der Arbeitsgruppe Pulmonalarterielle Hypertension der Österreichischen Gesellschaft für Lungenerkrankungen und Tuberkulose

Noch vor knapp 20 Jahren bedeutete die Diagnose „pulmonale Hypertension” ein Todesurteil für den betroffenen Patienten. Mit der Einführung von Prostazyklin als Therapeutikum in den frühen achtziger Jahren durch Timothy Higenbottam begann ein Prozess, der zur Fortentwicklung der medikamentösen Therapie der pulmonalarteriellen Hypertension und gleichzeitig zur fortschreitenden Erkenntnis der Grundlagen dieser Erkrankungsgruppe führte. Derzeit stehen vier verschiedene Prostaglandin-Derivate und ein Endothelin-Rezeptor-Antagonist zur Behandlung der pulmonalarteriellen Hypertension zur Verfügung. Dennoch – auch im Jahre 2003 sind Diagnose und Therapie der pulmonalarteriellen Hypertension nicht trivial. Die wichtigste Ursache hierfür ist, dass wir noch nicht verstehen, warum fundamental unterschiedliche Ursachen und Mechanismen einen sehr ähnlichen pathologischen Zustand, den eines funktionell fehlregulierten Gefäßwachstums, auslösen können. Trotz dieser fundamentalen biologischen Unkenntnis, die auch durch die ersten Daten zur genetischen Beeinflussung der familiären pulmonalen Hypertension nicht beseitigt wird, vermögen wir Patienten in immer besserer Weise zu helfen, zunehmend nicht nur durch die Anwendung eines Einzelwirkstoffes, sondern zum einen durch die Kombination unterschiedlicher Substanzen, und zum anderen durch gleichzeitige Gabe von Wirkstoffen auf unterschiedlichen Applikationswegen. Ein weiterer Grund für das ungenügende Verständnis der Pathogenese der Krankheit besteht darin, dass sich die pulmonale Hypertension längere Zeit nur durch sehr unspezifische Symptome manifestiert, die zudem im Symptomkomplex anderer, heute als „assoziiert“ eingeordneter Erkrankungen untergehen können. Dies hat für den Patienten häufig die fatale Folge, dass seine pulmonalarterielle Hypertension sehr spät oder zu spät erkannt wird. Vor diesem Hintergrund ist es ein Bestreben aller an der Behandlung von Patienten mit pulmonaler Hypertension Interessierten, ein nachvollziehbares einheitliches Vorgehen hinsichtlich Diagnose und Therapie der pulmonalen Hypertension zu erreichen, um eine möglichst rasche Verbesserung der bereits vorhandenen Therapieoptionen gewährleisten zu können. In diesem Sinne stellen die nachfolgend vorgestellten Empfehlungen der Arbeitsgruppe Pulmonale Hypertension der Österreichischen Gesellschaft für Lungenerkrankungen und Tuberkulose einen ersten Versuch dar, dieses Ziel zu realisieren. Es ist allen daran Beteiligten klar, dass diese Empfehlungen durch fortlaufende Diskussion verbessert werden können und sollten.

Middle age enhances expression of innate immunity genes in a female mouse model of pulmonary fibrosis.

The lungs are highly sensitive to tissue fibrosis, with a clear age-related component. Among the possible triggers of pulmonary fibrosis are repeated inhalations of fine organic particles. How age affects this response, is still far from being fully understood. We examined the impact of middle-age on gene expression in pulmonary fibrosis, using the novel “inhalation challenge set” mouse model. Our results demonstrate that the response of female mice to exposure of Pantoea agglomerans extract primarily involves various immune-related pathways and cell–cell/cell–extracellular matrix interactions. We found that middle-age had a strong effect on the response to the P. agglomerans-induced lung fibrosis, featured by a more rapid response and increased magnitude of expression changes. Genes belonging to innate immunity pathways (such as the TLR signaling and the NK-cell mediated cytotoxicity) were particularly up-regulated in middle-aged animals, suggesting that they may be potential targets for the treatment of pulmonary fibrosis caused by inhalations of organic particles. Our analysis also highlights the relevance of the “inhalation challenge set” mouse model to lung aging and related pathology.