397 results on '"Gazdhar, A"'
Search Results
202. Targeted Gene Transfer of Hepatocyte Growth Factor to Alveolar Type II Epithelial Cells Reduces Lung Fibrosis in Rats
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Gazdhar, Amiq, primary, Temuri, Almas, additional, Knudsen, Lars, additional, Gugger, Mathias, additional, Schmid, Ralph A., additional, Ochs, Matthias, additional, and Geiser, Thomas, additional
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- 2013
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203. Cell-specific expression of human HGF by alveolar type II cells induces remodeling of septal wall tissue in the lung: a morphometric study
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Leuenberger, Alexandra, primary, Gazdhar, Amiq, additional, Herrmann, Gudrun, additional, Ochs, Matthias, additional, Geiser, Thomas, additional, and Knudsen, Lars, additional
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- 2012
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204. An approach towards bronchoscopic-based gene therapy using electrical field accelerated plasmid droplets
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Hradetzky, D., primary, Boehringer, S., additional, Geiser, Th, additional, and Gazdhar, A., additional
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- 2012
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205. Time-lapse microscopy and classification of 2D human mesenchymal stem cells based on cell shape picks up myogenic from osteogenic and adipogenic differentiation
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Seiler, Christof, primary, Gazdhar, Amiq, additional, Reyes, Mauricio, additional, Benneker, Lorin M., additional, Geiser, Thomas, additional, Siebenrock, Klaus A., additional, and Gantenbein-Ritter, Benjamin, additional
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- 2012
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206. Induced Pluripotent Stem Cells (iPSC) Attenuate Fibrosis In Bleomycin Injured Rat Lungs
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Gazdhar, Amiq, primary, Grad, Iwona, additional, Gugger, Mathias, additional, Feki, Anis, additional, and Geiser, Thomas, additional
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- 2012
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207. Induzierbare pluripotente Stammzellen (iPSC) und deren Sekretionsprodukte vermindern die durch Bleomycin ausgelöste Lungenfibrose in Rattenlungen
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Gazdhar, A, primary, Grad, I, additional, Gugger, M, additional, Feki, A, additional, and Geiser, T, additional
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- 2012
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208. Microfluidic wound-healing assay to assess the regenerative effect of HGF on wounded alveolar epithelium
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Felder, Marcel, primary, Sallin, Pauline, additional, Barbe, Laurent, additional, Haenni, Beat, additional, Gazdhar, Amiq, additional, Geiser, Thomas, additional, and Guenat, Olivier, additional
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- 2012
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209. Improvement of Flap Necrosis in a Rat Random Skin Flap Model by In Vivo Electroporation-Mediated HGFGene Transfer
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Seyed Jafari, S. Morteza, Shafighi, Maziar, Beltraminelli, Helmut, Geiser, Thomas, Hunger, Robert E., and Gazdhar, Amiq
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- 2017
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210. Prolonged amelioration of acute lung allograft rejection by sequential overexpression of human interleukin-10 and hepatocyte growth factor in rats
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Fakin, Richard, primary, Hamacher, Jürg, additional, Gugger, Mathias, additional, Gazdhar, Amiq, additional, Moser, Helen, additional, and Alexander Schmid, Ralph, additional
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- 2011
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211. In Vivo Electroporation Mediated Gene Delivery to the Beating Heart
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Ayuni, Erick L., primary, Gazdhar, Amiq, additional, Giraud, Marie Noelle, additional, Kadner, Alexander, additional, Gugger, Mathias, additional, Cecchini, Marco, additional, Caus, Thierry, additional, Carrel, Thierry P., additional, Schmid, Ralph A., additional, and Tevaearai, Hendrik T., additional
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- 2010
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212. Effect of electroporation‐mediated diphtheria toxin A expression on PSA positive human prostate xenograft tumors in SCID mice
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Goepfert, Christine, primary, Gazdhar, Amiq, additional, Frey, Felix J., additional, and Frey, Brigitte M., additional
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- 2010
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213. Cellular Responses after Exposure of Lung Cell Cultures to Secondary Organic Aerosol Particles
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Gaschen, Annina, primary, Lang, Doris, additional, Kalberer, Markus, additional, Savi, Melanie, additional, Geiser, Thomas, additional, Gazdhar, Amiq, additional, Lehr, Claus-Michael, additional, Bur, Michael, additional, Dommen, Josef, additional, Baltensperger, Urs, additional, and Geiser, Marianne, additional
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- 2010
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214. Taurolidine in the prevention and therapy of lung metastases
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Hoksch, Beatrix, primary, Rufer, Benjamin, additional, Gazdhar, Amiq, additional, Bilici, Murat, additional, Beshay, Morris, additional, Gugger, Matthias, additional, and Schmid, Ralph Alexander, additional
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- 2009
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215. Comparison of procalcitonin and CrP in the postoperative course after lung decortication
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Carboni, Giovanni L., primary, Fahrner, Rene, additional, Gazdhar, Amiq, additional, Printzen, Gert, additional, Schmid, Ralph Alexander, additional, and Hoksch, Beatrix, additional
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- 2008
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216. Prolonged Amelioration of Acute Lung Allograft Rejection by Overexpression of Human Interleukin-10 Under Control of a Long Acting Ubiquitin C Promoter in Rats
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STAMMBERGER, U, primary, BILICI, M, additional, GUGGER, M, additional, GAZDHAR, A, additional, HAMACHER, J, additional, HYDE, S, additional, and SCHMID, R, additional
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- 2006
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217. 543. Monitoring Transgene Expression by Radio-Peptide Positron Emission Tomography
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Gazdhar, Amiq, primary, Bilici, Murat, additional, Ayuni, Erick, additional, Krause, Thomas, additional, Hofmann, Michal, additional, and Schmid, Raplh A., additional
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- 2006
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218. The Notch Signaling Pathway Is Related to Neurovascular Progression of Pancreatic Cancer
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B??chler, Peter, primary, Gazdhar, Amiq, additional, Schubert, Mario, additional, Giese, Nathalia, additional, Reber, Howard A., additional, Hines, Oscar J., additional, Giese, Thomas, additional, Ceyhan, G??ralp O., additional, M??ller, Michael, additional, B??chler, Markus W., additional, and Friess, Helmut, additional
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- 2005
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219. Synergistic effect of low dose Cyclosporine A and human interleukin 10 overexpression on acute rejection in rat lung allotransplantation
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PIEROG, J, primary, GAZDHAR, A, additional, STAMMBERGER, U, additional, GUGGER, M, additional, HYDE, S, additional, MATHIESEN, I, additional, GRODZKI, T, additional, and SCHMID, R, additional
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- 2005
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220. 925. Electroporation Mediated Gene Transfer of Human Interleukin-10 to Skeletal Muscle Reduces Acute Rejection in Rat Cardiac Allografts
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Jarosław Pieróg, Michele Genoni, Reza Tavakoli, Steven Hyde, Amiq Gazdhar, Anna Bagdannova, Ralph A. Schmid, and Mathias Gugger
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Pharmacology ,Electroporation ,Skeletal muscle ,Gene transfer ,Biology ,Molecular biology ,Interleukin 10 ,Transduction (genetics) ,medicine.anatomical_structure ,Plasmid dna ,Drug Discovery ,Genetics ,Cancer research ,medicine ,Molecular Medicine ,Molecular Biology ,Gene - Abstract
Background Electroporation-mediated transfer of plasmid DNA has been demonstrated to enhance gene transduction dramatically. The aim of this study was to investigate the impact of overexpression of human IL-10 (hIL-10) on acute rejection of cardiac allografts in the rat.
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- 2004
221. 512. Electroporation-Mediated Gene Transfer of Human Hepatocyte Growth Factor Reduces Bleomycin-Induced Pulmonary Fibrosis in Rats
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Ralph A. Schmid, Mathias Gugger, Amiq Gazdhar, Thomas Geiser, Steven Hyde, Patrick Fachinger, and Jarosław Pieróg
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medicine.medical_treatment ,Bleomycin ,Lung Disorder ,chemistry.chemical_compound ,Drug Discovery ,Pulmonary fibrosis ,Genetics ,medicine ,Molecular Biology ,Pharmacology ,biology ,business.industry ,Growth factor ,Electroporation ,respiratory system ,medicine.disease ,In vitro ,chemistry ,Mitogen-activated protein kinase ,Immunology ,Cancer research ,biology.protein ,Molecular Medicine ,Hepatocyte growth factor ,business ,medicine.drug - Abstract
Background: Pulmonary fibrosis may result in respiratory failure and a poor prognosis in patients with various types of interstitial lung disorders. Hepatocyte growth factor (HGF) is a potent mitogen for alveolar epithelial cells with antiapoptotic and fibrinolytic functions, resulting in improved alveolar repair in vitro. We hypothesized that HGF may improve pulmonary fibrosis in vitro by longterm expression of HGF using electroporation-mediated, non-viral gene transfer in the bleomycin-induced lung fibrosis model.
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- 2004
222. HEPATOLOGY Cartilage oligomeric matrix protein expression in hepatocellular carcinoma and the cirrhotic liver.
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Yi Xiao, Kleeff, Jörg, Junchao Guo, Jörg, Gazdhar, Amiq, Quan Liao, Amiq, di Cesare, Paul E, Büchler, Markus W., and Friess, Helmut
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THROMBOSPONDINS ,LIVER cancer ,CALCIUM-binding protein genes ,CARTILAGE diseases ,TUMOR growth ,CIRRHOSIS of the liver - Abstract
Cartilage oligomeric matrix protein (COMP) is the fifth member of the thrombospondin family of extracellular, calcium-binding proteins. It was initially isolated and characterized in cartilage tissue, where it is thought to contribute to the extracellular matrix composition and cell–extracellular matrix interaction. In the present study the expression of COMP was investigated in normal liver ( n = 19), liver cirrhosis ( n = 14) and hepatocellular carcinoma (HCC; n = 16) tissues, both at the mRNA and protein level. By northern blot and western blot analysis, COMP was absent or rarely expressed in the normal liver and liver cirrhosis tissues, but significantly overexpressed in HCC tissue samples. The COMP mRNA overexpression in HCC was not related to the clinical stage or tumor grade. By in situ hybridization and immunohistochemistry analysis, COMP mRNA and protein expression were localized within the cytoplasm of the tumor cells. COMP is highly expressed within the tumor cells of HCC, suggesting that COMP might play a role in the pathophysiology of this disease. © 2004 Blackwell Publishing Asia Pty Ltd [ABSTRACT FROM AUTHOR]
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- 2004
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223. Engineered nanoparticles for gene delivery in a bleomycin-injured lung fibrosis model
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von Garnier, C., Gazdhar, A., Fink, A., Hofmann, H., Gehr, P., Nicod, L. P., and Geiser, T.
224. Inhalational delivery of induced pluripotent stem cell secretome improves postpneumonectomy lung structure and function
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Connie C.W. Hsia, Yu-An Zhang, Thomas Geiser, Kemp H. Kernstine, Amiq Gazdhar, Khoa Cao, and D. Merrill Dane
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Physiology ,Angiogenesis ,medicine.medical_treatment ,Induced Pluripotent Stem Cells ,610 Medicine & health ,030204 cardiovascular system & hematology ,03 medical and health sciences ,Pneumonectomy ,Dogs ,0302 clinical medicine ,Physiology (medical) ,Diffusing capacity ,medicine ,Animals ,Humans ,Distribution (pharmacology) ,Induced pluripotent stem cell ,Intussusceptive angiogenesis ,Lung ,Inhalation ,business.industry ,respiratory system ,030104 developmental biology ,medicine.anatomical_structure ,Pulmonary Diffusing Capacity ,Female ,Lung Volume Measurements ,business ,Research Article - Abstract
Cell-free secretory products (secretome) of human induced pluripotent stem cells (iPSCs) have been shown to attenuate tissue injury and facilitate repair and recovery. To examine whether iPSC secretome facilitates mechanically induced compensatory responses following unilateral pneumonectomy (PNX), litter-matched young adult female hounds underwent right PNX (removing 55%–58% of lung units), followed by inhalational delivery of either the nebulized-conditioned media containing induced pluripotent stem cell secretome (iPSC CM) or control cell-free media (CFM); inhalation was repeated every 5 days for 10 treatments. Lung function was measured under anesthesia pre-PNX and 10 days after the last treatment (8 wk post-PNX); detailed quantitative analysis of lung ultrastructure was performed postmortem. Pre-PNX lung function was similar between groups. Compared with CFM control, treatment with iPSC CM attenuated the post-PNX decline in lung diffusing capacity for carbon monoxide and membrane diffusing capacity, accompanied by a 24% larger postmortem lobar volume and distal air spaces. Alveolar double-capillary profiles were 39% more prevalent consistent with enhanced intussusceptive angiogenesis. Frequency distribution of the harmonic mean thickness of alveolar blood-gas barrier shifted toward the lowest values, whereas alveolar septal tissue volume and arithmetic septal thickness were similar, indicating septal remodeling and reduced diffusive resistance of the blood-gas barrier. Thus, repetitive inhalational delivery of iPSC secretome enhanced post-PNX alveolar angiogenesis and septal remodeling that are associated with improved gas exchange compensation. Results highlight the plasticity of the remaining lung units following major loss of lung mass that are responsive to broad-based modulation provided by the iPSC secretome. NEW & NOTEWORTHY To examine whether the secreted products of human induced pluripotent stem cells (iPSCs) facilitate innate adaptive responses following loss of lung tissue, adult dogs underwent surgical removal of one lung, then received repeated administration of iPSC secretory products via inhalational delivery compared with control treatment. Inhalation of iPSC secretory products enhanced capillary formation and beneficial structural remodeling in the remaining lung, leading to improved lung function.
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225. Mitochondrial DNA mutations and respiratory chain dysfunction in idiopathic and connective tissue disease-related lung fibrosis
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Jaeger, Veronika K, Lebrecht, Dirk, Nicholson, Andrew G, Wells, Athol, Bhayani, Harshil, Gazdhar, Amiq, Tamm, Michael, Venhoff, Nils, Geiser, Thomas, and Walker, Ulrich A
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respiratory system ,610 Medicine & health ,respiratory tract diseases ,3. Good health - Abstract
Reactive oxygen species (ROS) are implicated in the aetiology of interstitial lung disease (ILD). We investigated the role of large-scale somatically acquired mutations in mitochondrial DNA (mtDNA) and consecutive respiratory chain dysfunction as a trigger of ROS-formation and lung fibrosis. Mitochondria were analysed in lung biopsies from 30 patients with idiopathic or connective tissue disease (CTD)-related ILD and 13 controls. In 17 patients we had paired biopsies from upper and lower lobes. Control samples were taken from lung cancer resections without interstitial fibrosis. Malondialdehyde, a marker of ROS-formation, was elevated in ILD-biopsies (p = 0.044). The activity of the mitochondrial respiratory chain (cytochrome c-oxidase/succinate dehydrogenase [COX/SDH]-ratio) was depressed in ILD (median = 0.10,) compared with controls (0.12, p
226. The secretome of induced pluripotent stem cells reduces lung fibrosis in part by hepatocyte growth factor
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Tamò, Luca, Feki, Anis, Gugger, Mathias, Gazdhar, Amiq Ur Rahman, Geiser, Thomas, and Grad, I
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respiratory system ,610 Medicine & health ,3. Good health - Abstract
INTRODUCTION Idiopathic pulmonary fibrosis (IPF) is a progressive and irreversible fibrotic lung disease, resulting in respiratory insufficiency and reduced survival. Pulmonary fibrosis is a result of repeated alveolar epithelial microinjuries, followed by abnormal regeneration and repair processes in the lung. Recently, stem cells and their secretome have been investigated as a novel therapeutic approach in pulmonary fibrosis. We evaluated the potential of induced pluripotent stem cells (iPSC) conditioned media (iPSC-cm) to regenerate and repair the alveolar epithelium in vitro and improve bleomycin induced lung injury in vivo. METHODS IPSC-cm was collected from cultured iPSC derived from human foreskin fibroblasts and its biological effects on alveolar epithelial wound repair was studied in an alveolar wound healing assay in vitro. Furthermore, iPSC-cm was intratracheally instilled 7 days after bleomycin induced injury in the rat lungs and histologically and biochemically assessed 7 days after instillation. RESULTS iPSC-cm increased alveolar epithelial wound repair in vitro compared with medium control. Intratracheal instillation of iPSC-cm in bleomycin-injured lungs reduced the collagen content and improved lung fibrosis in the rat lung in vivo. Profibrotic TGFbeta1 and alpha-smooth muscle actin (alpha-sma) expression were markedly reduced in the iPSC-cm treated group compared with control. Antifibrotic hepatocyte growth factor (HGF) was detected in iPSC-cm in biologically relevant levels, and specific inhibition of HGF in iPSC-cm attenuated the antifibrotic effect of iPSC-cm, indicating a central role of HGF in iPSC-cm. CONCLUSION iPSC-cm increased alveolar epithelial wound repair in vitro and attenuated bleomycin induced fibrosis in vivo, partially due to the presence of HGF and may represent a promising novel, cell free therapeutic option against lung injury and fibrosis.
227. Generation of an alveolar epithelial type II cell line from induced pluripotent stem cells
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Tamò, Luca, Hibaoui, Youssef, Kallol, Sampada, Alves, Marco, Albrecht, Christiane, Hostettler, Katrin E, Feki, Anis, Rougier, Jean-Sébastien, Abriel, Hugues, Knudsen, Lars, Gazdhar, Amiq, and Geiser, Thomas
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630 Agriculture ,570 Life sciences ,biology ,respiratory system ,610 Medicine & health ,3. Good health - Abstract
Differentiation of primary alveolar type II epithelial cells (AEC type II) to AEC type I in culture is a major barrier in the study of the alveolar epithelium in vitro. The establishment of an AEC type II cell line derived from induced pluripotent stem cells (iPSC) represents a novel opportunity to study alveolar epithelial cell biology, for instance in the context of lung injury, fibrosis and repair. In the present study, we generated long lasting AECII from iPSC (iPSC-AEC). Long lasting iPSC-AECII (LL)-iPSC-AECII) displayed morphological characteristics of AECII including growth in a cobble stone monolayer, the presence of lamellar bodies and microvilli as shown by electron microscopy. Also, (LL)-iPSC-AECII expressed AEC type II proteins such as cytokeratin, surfactant protein C and lysotracker DND 26 (a marker for lamellar bodies). Furthermore, the (LL)-iPSC-AECII exhibited functional properties of AECII by an increase of transepithelial electrical resistance (TERR) over time, secretion of inflammatory mediators in biologically relevant quantities (interleukin-6, interleukin-8) and efficient in vitro alveolar epithelial wound repair. Consistent with the AECII phenotype, the cell line showed the ability to uptake and release surfactant protein B, to secrete phospholipids and to differentiate into AEC type I. In summary, we established a long lasting, but finite AEC type II cell line derived from iPSC as a novel cellular model to study alveolar epithelial cell biology in lung health and disease.
228. Synergistic effect of low dose Cyclosporine A and human interleukin 10 overexpression on acute rejection in rat lung allotransplantation
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Pierog, Jaroslaw, Gazdhar, Amiq, Stammberger, Uz, Gugger, Matthias, Hyde, Steven, Mathiesen, Iacob, Grodzki, Tomasz, Schmid, Ralph A., Pierog, Jaroslaw, Gazdhar, Amiq, Stammberger, Uz, Gugger, Matthias, Hyde, Steven, Mathiesen, Iacob, Grodzki, Tomasz, and Schmid, Ralph A.
- Abstract
Objective: Electroporation mediated transfer of plasmid DNA into peripheral muscle results in high transfection efficiency. The aim of this study was to investigate the effect of gene transfer of human IL-10 (hIL-10) into the tibialis anterior muscle (MTA) in combination with low dose Cyclosporine A (CsA) on acute rejection of lung allografts in the rat. Methods: Lung allotransplantation was performed from male BN donor to male Fisher F344 rats. Gene transfer was achieved by intramuscular injection into the MTA of the recipient followed by electroporation (4×20ms impulses at 200V/cm) 24h prior to the transplantation. Group A (n=5) received CsA (2.5mg/kg bw ip) for 5 days post-transplant and group B (n=5) 2.5μg of PCIK hIL-10 (plasmid expression vector containing human CMV immediate early gene promoter and enhancer) and a low dose CsA (2.5mg/kg bw i.p.). Graft function was assessed by blood gas at day 5 after exclusion of the native lung. Animals were sacrificed and blood was drawn to measure serum hIL-10 levels (ELISA) and tissue was sampled for histological grading of rejection. Results: Local expression of hIL-10 was confirmed at the mRNA level by in situ hybridization. All group A control animals showed severe signs of rejection. At day 5 all grafts in group B showed good gas exchange mean PaO2 233±123mmHg, vs 44±8mmHg in group A. Histological examination revealed moderate to severe rejection in all animals in group A (IIIB, ISHLT) in contrast to low moderate rejection in group B (II-IIIA). hIL-10 serum levels on day 5 were 14±7pg/ml in group B vs. 0 in group A. Conclusions: Electroporation mediated hIL-10 overexpression in a peripheral muscle of the recipient in combination with low dose CsA reduces acute rejection in this model of rat lung allotransplantation
229. Taurolidine in the prevention and therapy of lung metastases
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Hoksch, Beatrix, Rufer, Benjamin, Gazdhar, Amiq, Bilici, Murat, Beshay, Morris, Gugger, Matthias, Schmid, Ralph Alexander, Hoksch, Beatrix, Rufer, Benjamin, Gazdhar, Amiq, Bilici, Murat, Beshay, Morris, Gugger, Matthias, and Schmid, Ralph Alexander
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Objective: During surgery for colon carcinoma, tumour cells may spread into the blood and may lead to the development of distant metastases. The most frequent sites of metastases are the liver and lungs. A new therapeutic approach is required to prevent tumour implantation of freely circulating tumour cells during and after surgery and to treat established metastases. The aim of this prospective study was to observe the influence of long-term intravenous taurolidine on the development of lung metastases after intravenous injection of colon adenocarcinoma cells. Methods: Tumour cells (DHD/K12/TRb colon adenocarcinoma cell line, 1×106 cells) were injected into the right vena jugularis interna of BDIX rats. The animals (n=13) were randomised into three groups: group 1: tumour cell implantation without taurolidine application (control group); group 2: tumour cell implantation and simultaneous start of the taurolidine injection through osmotic pump, removal of the osmotic pump on day 7; group 3: tumour cell implantation on day 0 and start of the taurolidine injection through osmotic pump on day 14. Results: In the taurolidine groups, the number and size of lung metastases were significantly lower compared to the control group (p=0.018; p=0.018 and p=0.036; p=0.018). Although the results of the intravenous long-term therapy with taurolidine in group 2 did not reach statistical significance in comparison with the results of group 3, a positive trend was revealed: The mean number of metastases in group 2 was 18.2 versus 28.2 in group 3. Conclusions: The application of taurolidine tends to prevent the development of lung metastases. Furthermore, taurolidine seems to reduce established lung metastases in this in vivo model. Taurolidine may offer additional therapeutic options in patients with colon adenocarcinoma
230. Design of Miniaturized Electro Spray Instrument for Gene and Drug Therapeutic Treatment of IPF
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Fiave, Prosper Agbesi, Boehringer, Stephan, Gazdhar, Amiq Ur Rahman, Geiser, Thomas, Hradetzky, David, Fiave, Prosper Agbesi, Boehringer, Stephan, Gazdhar, Amiq Ur Rahman, Geiser, Thomas, and Hradetzky, David
231. Comparison of procalcitonin and CrP in the postoperative course after lung decortication
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Carboni, Giovanni L., Fahrner, Rene, Gazdhar, Amiq, Printzen, Gert, Schmid, Ralph Alexander, Hoksch, Beatrix, Carboni, Giovanni L., Fahrner, Rene, Gazdhar, Amiq, Printzen, Gert, Schmid, Ralph Alexander, and Hoksch, Beatrix
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Objective: The objective of this prospective study was to compare the clinical value of procalcitonin (PCT) and C-reactive protein (CrP) plasma concentrations in their postoperative course after decortication. Methods: Twenty-two patients requiring surgery for pleural empyema were chosen for this prospective study. Routine blood samples including CrP and PCT plasma concentrations were taken before the operation and on the 1st, 2nd, 3rd, and 7th postoperative day. Results: Due to infection PCT and CrP were elevated preoperatively. In the postoperative course both PCT and CrP reached peak-levels on day 2 with values up to 43.55ng/ml and 384.00mg/l, respectively. In PCT the rise was followed by a clear decrease in 20 (90.9 %) patients until day 7. In contrast the CrP levels decreased slowly and only seven (54.5%) patients had values of 100mg/l or below on day 7. PCT showed a better correlation with the clinic in case of septic course than CrP does. Conclusions: PCT reflects postoperative clinical course more accurately than CrP. Therefore, PCT is a more appropriate laboratory parameter to monitor patients after surgery for pleural empyema
232. Size-dependent accumulation of particles in lysosomes modulates dendritic cell function through impaired antigen degradation
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Seydoux, Emilie, Rothen-Rutishauser, Barbara, Nita, Izabela M, Balog, Sandor, Gazdhar, Amiq, Stumbles, Philip A, Petri-Fink, Alke, Blank, Fabian, Garnier, Christophe von, Seydoux, Emilie, Rothen-Rutishauser, Barbara, Nita, Izabela M, Balog, Sandor, Gazdhar, Amiq, Stumbles, Philip A, Petri-Fink, Alke, Blank, Fabian, and Garnier, Christophe von
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Introduction: Nanosized particles may enable therapeutic modulation of immune responses by targeting dendritic cell (DC) networks in accessible organs such as the lung. To date, however, the effects of nanoparticles on DC function and downstream immune responses remain poorly understood. Methods: Bone marrow–derived DCs (BMDCs) were exposed in vitro to 20 or 1,000 nm polystyrene (PS) particles. Particle uptake kinetics, cell surface marker expression, soluble protein antigen uptake and degradation, as well as in vitro CD4⁺ T-cell proliferation and cytokine production were analyzed by flow cytometry. In addition, co-localization of particles within the lysosomal compartment, lysosomal permeability, and endoplasmic reticulum stress were analyzed. Results: The frequency of PS particle–positive CD11c⁺/CD11b⁺ BMDCs reached an early plateau after 20 minutes and was significantly higher for 20 nm than for 1,000 nm PS particles at all time-points analyzed. PS particles did not alter cell viability or modify expression of the surface markers CD11b, CD11c, MHC class II, CD40, and CD86. Although particle exposure did not modulate antigen uptake, 20 nm PS particles decreased the capacity of BMDCs to degrade soluble antigen, without affecting their ability to induce antigen-specific CD4⁺ T-cell proliferation. Co-localization studies between PS particles and lysosomes using laser scanning confocal microscopy detected a significantly higher frequency of co-localized 20 nm particles as compared with their 1,000 nm counterparts. Neither size of PS particle caused lysosomal leakage, expression of endoplasmic reticulum stress gene markers, or changes in cytokines profiles. Conclusion: These data indicate that although supposedly inert PS nanoparticles did not induce DC activation or alteration in CD4⁺ T-cell stimulating capacity, 20 nm (but not 1,000 nm) PS particles may reduce antigen degradation through interference
233. A Case of Idiopathic Tetanus Treated with Sub-Cutaneous Injection of EserineSulphas: Recovery
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Gazdhar, Sorabji Furdoonji
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A Mirror of Hospital Practice - Published
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234. 249: Prolonged amelioration of acute lung allograft rejection by overexpression of human interleukin-10 under control of a long acting ubiquitin C promoter in rats
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Stammberger, U., Bilici, M., Gazdhar, A., and Schmid, R.A.
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- 2006
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235. Interactome Analysis of iPSC Secretome and Its Effect on Macrophages In Vitro.
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Tamò, Luca, Fytianos, Kleanthis, Caldana, Fabienne, Simillion, Cedric, Feki, Anis, Nita, Izabela, Heller, Manfred, Geiser, Thomas, and Gazdhar, Amiq
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INDUCED pluripotent stem cells , *AMYLOID beta-protein precursor , *MACROPHAGES , *BLOOD cells , *PULMONARY fibrosis - Abstract
Simple Summary: Macrophages play essential role in repair, regeneration and tissue remodeling. Role of macrophages in progression of lung fibrosis is established. Secretome of Induced pluripotent stem cells (iPSC-CM) has shown to reduce lung fibrosis and regulate macrophage phenotype, however exact mechanism is not known. Using advanced bioinformatics analysis by gene network analysis in this study we identified two components AAP and ELAVL-1 present in the iPSC-CM playing important role in regulation of macrophage phenotype. In this invitro study we confirmed experimentally that AAP and ELAVL1 play essential role by changing the profibrotic phenotype of the macrophages to pro resolution macrophages. We demonstrate reduction in gene expression and cytokine secretion of profibrotic macrophages after iPSC-CM treatment. Our study confirms antifibrotic and regenerative potential of iPSC-CM. Induced pluripotent stem cell secretome (iPSC-CM) mitigate organ injury and help in repair. Macrophages play a critical role in tissue repair and regeneration and can be directed to promote tissue repair by iPSC-CM, although the exact mechanisms are not known. In the current investigative study, we evaluated the possible mechanism by which iPSC-CM regulates the phenotype and secretory pattern of macrophages in vitro. Macrophages were obtained from human peripheral blood mononuclear cells and differentiated to various subpopulations and treated with either iPSC-CM or control media in vitro. Macrophage phenotype was assessed by flow cytometry, gene expression changes by qRT PCR and secretory pattern by multiplex protein analysis. The protein and gene interaction network revealed the involvement of Amyloid precursor protein (APP) and ELAV-like protein 1 (ELAVL-1) both present in the iPSC-CM to play an important role in regulating the macrophage phenotype and their secretory pattern. This exploratory study reveals, in part, the possible mechanism and identifies two potential targets by which iPSC-CM regulate macrophages and help in repair and regeneration. [ABSTRACT FROM AUTHOR]
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- 2021
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236. In Vivo Electroporation-Mediated, Intrahepatic Alpha1 Antitrypsin Gene Transfer Reduces Pulmonary Emphysema in Pallid Mice.
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Sutter, Marco A., Cremona, Tiziana P., Nita, Izabela, Cavarra, Eleonora, Lungarella, Giuseppe, Lewis, Eli C., Schittny, Johannes C., Geiser, Thomas, and Gazdhar, Amiq
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PULMONARY emphysema , *GENETIC transformation , *ELECTROPORATION , *TRYPSIN inhibitors , *HUMAN genes , *LUNGS - Abstract
Rationale: Mutation in the alpha1 antitrypsin (AAT) gene leads to low circulating levels of AAT, which is associated with several disease processes including pulmonary emphysema. The standard of care relies on substitution with plasma-purified AAT. We studied a novel approach to obtain sustained therapeutic levels of circulating AAT using nonviral in vivo electroporation-mediated gene transfer to the liver. Methods: In vivo intrahepatic electroporation-mediated human AAT gene transfer was performed in C57 Bl/6J mice carrying a genetic deficiency of murine AAT (pallid mice) and suffering from pulmonary emphysema. The animals were evaluated for lung function using flexiVent and detailed stereological assessments. Lung neutrophilic burden was assessed. Results: Pallid mice showed morphologically detectable pulmonary emphysema. Thirty days after in vivo electroporation-mediated gene transfer directly aimed at the liver, circulating human AAT was elevated and lung function was significantly improved compared to non-treated pallid mice. Stereological analysis revealed a reduction in pulmonary emphysema. Conclusion: Our data indicate that in vivo intrahepatic electroporation-mediated gene transfer of AAT is a safe and efficient procedure resulting in reduction of pulmonary emphysema in pallid mice. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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237. Mitochondrial DNA mutations and respiratory chain dysfunction in idiopathic and connective tissue disease-related lung fibrosis.
- Author
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Jaeger, Veronika K., Lebrecht, Dirk, Nicholson, Andrew G., Wells, Athol, Bhayani, Harshil, Gazdhar, Amiq, Tamm, Michael, Venhoff, Nils, Geiser, Thomas, and Walker, Ulrich A.
- Abstract
Reactive oxygen species (ROS) are implicated in the aetiology of interstitial lung disease (ILD). We investigated the role of large-scale somatically acquired mutations in mitochondrial DNA (mtDNA) and consecutive respiratory chain dysfunction as a trigger of ROS-formation and lung fibrosis. Mitochondria were analysed in lung biopsies from 30 patients with idiopathic or connective tissue disease (CTD)-related ILD and 13 controls. In 17 patients we had paired biopsies from upper and lower lobes. Control samples were taken from lung cancer resections without interstitial fibrosis. Malondialdehyde, a marker of ROS-formation, was elevated in ILD-biopsies (p = 0.044). The activity of the mitochondrial respiratory chain (cytochrome c-oxidase/succinate dehydrogenase [COX/SDH]-ratio) was depressed in ILD (median = 0.10,) compared with controls (0.12, p < 0.001), as was the expression of mtDNA-encoded COX-subunit-2 protein normalized for the nucleus-encoded COX-subunit-4 (COX2/COX4-ratio; ILD-median = 0.6; controls = 2.2; p < 0.001). Wild-type mtDNA copies were slightly elevated in ILD (p = 0.088). The common mtDNA deletion was only present at low levels in controls (median = 0%) and at high levels in ILD (median = 17%; p < 0.001). In ILD-lungs with paired biopsies, lower lobes contained more malondialdehyde and mtDNA deletions than upper lobes and had lower COX2/COX4-ratios and COX/SDH-ratios (all p < 0.001). Acquired mtDNA-mutations and consecutive respiratory chain dysfunction may both trigger and perpetuate ROS-formation in ILD. [ABSTRACT FROM AUTHOR]
- Published
- 2019
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238. Influence of Microenvironmental Orchestration on Multicellular Lung Alveolar Organoid Development from Human Induced Pluripotent Stem Cells.
- Author
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Ozan VB, Wang H, Akshay A, Anand D, Hibaoui Y, Feki A, Gote-Schniering J, Gheinani AH, Heller M, Uldry AC, Lagache SB, Gazdhar A, and Geiser T
- Abstract
Induced pluripotent stem cells (iPSCs) have emerged as promising in vitro tools, providing a robust system for disease modelling and facilitating drug screening. Human iPSCs have been successfully differentiated into lung cells and three-dimensional lung spheroids or organoids. The lung is a multicellular complex organ that develops under the symphonic influence of the microenvironment. Here, we hypothesize that the generation of lung organoids in a controlled microenvironment (cmO) (oxygen and pressure) yields multicellular organoids with architectural complexity resembling the lung alveoli. iPSCs were differentiated into mature lung organoids following a stepwise protocol in an oxygen and pressure-controlled microenvironment. The organoids developed in the controlled microenvironment displayed complex alveolar architecture and stained for SFTPC, PDPN, and KRT5, indicating the presence of alveolar epithelial type II and type I cells, as well as basal cells. Moreover, gene and protein expression levels were also increased in the cmO. Furthermore, pathway analysis of proteomics revealed upregulation of lung development-specific pathways in the cmO compared to those growing in normal culture conditions. In summary, by using a controlled microenvironment, we established a complex multicellular lung organoid derived from iPSCs as a novel cellular model to study lung alveolar biology in both lung health and disease., (© 2024. The Author(s).)
- Published
- 2024
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239. Biogenic polymer-based patches for congenital cardiac surgery: a feasibility study.
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Richert E, Nienhaus A, Ekroll Jahren S, Gazdhar A, Grab M, Hörer J, Carrel T, Obrist D, and Heinisch PP
- Abstract
Objective: Currently used patch materials in congenital cardiac surgery do not grow, renew, or remodel. Patch calcification occurs more rapidly in pediatric patients eventually leading to reoperations. Bacterial cellulose (BC) as a biogenic polymer offers high tensile strength, biocompatibility, and hemocompatibility. Thus, we further investigated the biomechanical properties of BC for use as patch material., Methods: The BC-producing bacteria Acetobacter xylinum were cultured in different environments to investigate optimal culturing conditions. For mechanical characterization, an established method of inflation for biaxial testing was used. The applied static pressure and deflection height of the BC patch were measured. Furthermore, a displacement and strain distribution analysis was performed and compared to a standard xenograft pericardial patch., Results: The examination of the culturing conditions revealed that the BC became homogenous and stable when cultivated at 29°C, 60% oxygen concentration, and culturing medium exchange every third day for a total culturing period of 12 days. The estimated elastic modulus of the BC patches ranged from 200 to 530 MPa compared to 230 MPa for the pericardial patch. The strain distributions, calculated from preloaded (2 mmHg) to 80 mmHg inflation, show BC patch strains ranging between 0.6% and 4%, which was comparable to the pericardial patch. However, the pressure at rupture and peak deflection height varied greatly, ranging from 67 to around 200 mmHg and 0.96 to 5.28 mm, respectively. The same patch thickness does not automatically result in the same material properties indicating that the manufacturing conditions have a significant impact on durability., Conclusions: BC patches can achieve comparable results to pericardial patches in terms of strain behavior as well as in the maximum applied pressure that can be withstood without rupture. Bacterial cellulose patches could be a promising material worth further research., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (© 2023 Richert, Nienhaus, Ekroll Jahren, Gazdhar, Grab, Hörer, Carrel, Obrist and Heinisch.)
- Published
- 2023
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240. Mesenchyme-derived vertebrate lonesome kinase controls lung organogenesis by altering the matrisome.
- Author
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Brütsch SM, Madzharova E, Pantasis S, Wüstemann T, Gurri S, Steenbock H, Gazdhar A, Kuhn G, Angel P, Bellusci S, Brinckmann J, Auf dem Keller U, Werner S, and Bordoli MR
- Subjects
- Animals, Mice, Organogenesis genetics, Lung, Mesoderm, Vertebrates, Protein-Tyrosine Kinases, Protein Kinases genetics, Extracellular Matrix
- Abstract
Vertebrate lonesome kinase (VLK) is the only known secreted tyrosine kinase and responsible for the phosphorylation of a broad range of secretory pathway-resident and extracellular matrix proteins. However, its cell-type specific functions in vivo are still largely unknown. Therefore, we generated mice lacking the VLK gene (protein kinase domain containing, cytoplasmic (Pkdcc)) in mesenchymal cells. Most of the homozygous mice died shortly after birth, most likely as a consequence of their lung abnormalities and consequent respiratory failure. E18.5 embryonic lungs showed a reduction of alveolar type II cells, smaller bronchi, and an increased lung tissue density. Global mass spectrometry-based quantitative proteomics identified 97 proteins with significantly and at least 1.5-fold differential abundance between genotypes. Twenty-five of these had been assigned to the extracellular region and 15 to the mouse matrisome. Specifically, fibromodulin and matrilin-4, which are involved in extracellular matrix organization, were significantly more abundant in lungs from Pkdcc knockout embryos. These results support a role for mesenchyme-derived VLK in lung development through regulation of matrix dynamics and the resulting modulation of alveolar epithelial cell differentiation., (© 2023. The Author(s).)
- Published
- 2023
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241. Anti-Fibrotic Effect of SDF-1β Overexpression in Bleomycin-Injured Rat Lung.
- Author
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Fytianos K, Schliep R, Mykoniati S, Khan P, Hostettler KE, Tamm M, Gazdhar A, Knudsen L, and Geiser T
- Abstract
Rational: Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease and is associated with high mortality due to a lack of effective treatment. Excessive deposition of the extracellular matrix by activated myofibroblasts in the alveolar space leads to scar formation that hinders gas exchange. Therefore, selectively removing activated myofibroblasts with the aim to repair and remodel fibrotic lungs is a promising approach. Stromal-derived growth factor (SDF-1) is known to stimulate cellular signals which attract stem cells to the site of injury for tissue repair and remodeling. Here, we investigate the effect of overexpression of SDF-1β on lung structure using the bleomycin-injured rat lung model. Methods: Intratracheal administration of bleomycin was performed in adult male rats (F344). Seven days later, in vivo electroporation-mediated gene transfer of either SDF-1β or the empty vector was performed. Animals were sacrificed seven days after gene transfer and histology, design-based stereology, flow cytometry, and collagen measurement were performed on the tissue collected. For in vitro experiments, lung fibroblasts obtained from IPF patients were used. Results: Seven days after SDF-1β gene transfer to bleomycin-injured rat lungs, reduced total collagen, reduced collagen fibrils, improved histology and induced apoptosis of myofibroblasts were observed. Furthermore, it was revealed that TNF-α mediates SDF-1β-induced apoptosis of myofibroblasts; moreover, SDF-1β overexpression increased alveolar epithelial cell numbers and proliferation in vivo and also induced their migration in vitro. Conclusions: Our study demonstrates a new antifibrotic mechanism of SDF-1β overexpression and suggests SDF-1β as a potential new approach for the treatment of lung fibrosis.
- Published
- 2022
- Full Text
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242. Basal-Like Cell-Conditioned Medium Exerts Anti-Fibrotic Effects In Vitro and In Vivo .
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Khan P, Fytianos K, Blumer S, Roux J, Gazdhar A, Savic S, Knudsen L, Jonigk D, Kuehnel MP, Mykoniati S, Tamm M, Geiser T, and Hostettler KE
- Abstract
In idiopathic pulmonary fibrosis (IPF), basal-like cells are atypically present in the alveolar region, where they may affect adjacent stromal cells by paracrine mechanisms. We here aimed to confirm the presence of basal-like cells in peripheral IPF lung tissue in vivo , to culture and characterize the cells in vitro, and to investigate their paracrine effects on IPF fibroblasts in vitro and in bleomycin-injured rats in vivo . Basal-like cells are mainly localized in areas of pathological bronchiolization or honeycomb cysts in peripheral IPF lung tissue. Single-cell RNA sequencing (scRNA-seq) demonstrated an overall homogeneity, the expression of the basal cell markers cytokeratin KRT5 and KRT17, and close transcriptomic similarities to basal cells in the majority of cells cultured in vitro . Basal-like cells secreted significant levels of prostaglandin E2 (PGE2), and their conditioned medium (CM) inhibited alpha-smooth muscle actin (α-SMA) and collagen 1A1 (Col1A1) and upregulated matrix metalloproteinase-1 (MMP-1) and hepatocyte growth factor (HGF) by IPF fibroblasts in vitro . The instillation of CM in bleomycin-injured rat lungs resulted in reduced collagen content, improved lung architecture, and reduced α-SMA-positive cells. Our data suggested that basal-like cells may limit aberrant fibroblast activation and differentiation in IPF through paracrine mechanisms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Khan, Fytianos, Blumer, Roux, Gazdhar, Savic, Knudsen, Jonigk, Kuehnel, Mykoniati, Tamm, Geiser and Hostettler.)
- Published
- 2022
- Full Text
- View/download PDF
243. LRR-protein RNH1 dampens the inflammasome activation and is associated with COVID-19 severity.
- Author
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Bombaci G, Sarangdhar MA, Andina N, Tardivel A, Yu EC, Mackie GM, Pugh M, Ozan VB, Banz Y, Spinetti T, Hirzel C, Youd E, Schefold JC, Taylor G, Gazdhar A, Bonadies N, Angelillo-Scherrer A, Schneider P, Maslowski KM, and Allam R
- Subjects
- Animals, COVID-19 immunology, Caspase 1 metabolism, Humans, Male, Mice, Mice, Inbred C57BL, Mice, Knockout, NF-kappa B metabolism, NLR Family, Pyrin Domain-Containing 3 Protein metabolism, Patient Acuity, Proteasome Endopeptidase Complex metabolism, COVID-19 pathology, Carrier Proteins metabolism, Inflammasomes metabolism, Leucine-Rich Repeat Proteins metabolism
- Abstract
Inflammasomes are cytosolic innate immune sensors of pathogen infection and cellular damage that induce caspase-1-mediated inflammation upon activation. Although inflammation is protective, uncontrolled excessive inflammation can cause inflammatory diseases and can be detrimental, such as in coronavirus disease (COVID-19). However, the underlying mechanisms that control inflammasome activation are incompletely understood. Here we report that the leucine-rich repeat (LRR) protein ribonuclease inhibitor (RNH1), which shares homology with LRRs of NLRP (nucleotide-binding oligomerization domain, leucine-rich repeat, and pyrin domain containing) proteins, attenuates inflammasome activation. Deletion of RNH1 in macrophages increases interleukin (IL)-1β production and caspase-1 activation in response to inflammasome stimulation. Mechanistically, RNH1 decreases pro-IL-1β expression and induces proteasome-mediated caspase-1 degradation. Corroborating this, mouse models of monosodium urate (MSU)-induced peritonitis and lipopolysaccharide (LPS)-induced endotoxemia, which are dependent on caspase-1, respectively, show increased neutrophil infiltration and lethality in Rnh1 mice compared with wild-type mice. Furthermore, RNH1 protein levels were negatively related with disease severity and inflammation in hospitalized COVID-19 patients. We propose that RNH1 is a new inflammasome regulator with relevance to COVID-19 severity.
-/- mice compared with wild-type mice. Furthermore, RNH1 protein levels were negatively related with disease severity and inflammation in hospitalized COVID-19 patients. We propose that RNH1 is a new inflammasome regulator with relevance to COVID-19 severity., (© 2022 Bombaci et al.)- Published
- 2022
- Full Text
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244. Electrospray Mediated Localized and Targeted Chemotherapy in a Mouse Model of Lung Cancer.
- Author
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Ruzgys P, Böhringer S, Dokumaci AS, Hari Y, Schürch CM, Brühl F, Schürch S, Szidat S, Riether C, Šatkauskas S, Geiser T, Hradetzky D, and Gazdhar A
- Abstract
Background: An advanced stage, centrally localized invasive tumor is a major cause of sudden death in lung cancer patients. Currently, chemotherapy, radiotherapy, laser ablation, or surgical resection if possible are the available state-of-the-art treatments but none of these guarantee remedy or long-term relief and are often associated with fatal complications. Allowing localized chemotherapy, by direct and confined drug delivery only at the tumor site, could be a promising option for preoperative down staging or palliative therapy. Here we report the localized and targeted application of intra tumor delivery of chemotherapeutics using a novel device based on the principle of electrospray. Methods: C57BL/6J mice were injected with Lewis lung carcinoma cells subcutaneously. After 15 days, the animals were anesthetized and the tumors were exposed by skin incision. Tumors were electrosprayed with 100 µg cisplatin on days 0 and 2, and tumor volumes were measured daily. Animals were sacrificed on day 7 after the first electrospray and tumors were analyzed by immunohistochemistry. Results: In this proof-of-concept study, we report that the tumor volume was reduced by 81.2% (22.46 ± 12.14 mm
3 ) after two electrospray mediated Cisplatin deliveries, while the control tumor growth, at the same time point, increased by 200% (514.30 ± 104.50 mm3 ). Moreover, tunnel and Caspase-3 positive cells were increased after Cisplatin electrospray compared to other experimental groups of animals. Conclusion: Targeted drug delivery by electrospray is efficient in the subcutaneous mouse model of lung cancer and offers a promising opportunity for further development toward its clinical application., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Ruzgys, Böhringer, Dokumaci, Hari, Schürch, Brühl, Schürch, Szidat, Riether, Šatkauskas, Geiser, Hradetzky and Gazdhar.)- Published
- 2021
- Full Text
- View/download PDF
245. Efficacy of Combined in-vivo Electroporation-Mediated Gene Transfer of VEGF, HGF, and IL-10 on Skin Flap Survival, Monitored by Label-Free Optical Imaging: A Feasibility Study.
- Author
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Seyed Jafari SM, Blank F, Ramser HE, Woessner AE, Shafighi M, Geiser T, Quinn KP, Hunger RE, and Gazdhar A
- Abstract
Preventing surgical flaps necrosis remains challenging. Laser Doppler imaging and ultrasound can monitor blood flow in flap regions, but they do not directly measure the cellular response to ischemia. The study aimed to investigate the efficacy of synergistic in-vivo electroporation-mediated gene transfer of interleukin 10 (IL-10) with either hepatocyte growth factor (HGF) or vascular endothelial growth factor (VEGF) on the survival of a modified McFarlane flap, and to evaluate the effect of the treatment on cell metabolism, using label-free fluorescence lifetime imaging. Fifteen male Wistar rats (290-320 g) were randomly divided in three groups: group-A (control group) underwent surgery and received no gene transfer. Group-B received electroporation mediated hIL-10 gene delivery 24 h before and VEGF gene delivery 24 h after surgery. Group-C received electroporation mediated hIL-10 gene delivery 24 h before and hHGF gene delivery 24 h after surgery. The animals were assessed clinically and histologically. In addition, label-free fluorescence lifetime imaging was performed on the flap. Synergistic electroporation mediated gene delivery significantly decreased flap necrosis ( P = 0.0079) and increased mean vessel density ( P = 0.0079) in treatment groups B and C compared to control group-A. NADH fluorescence lifetime analysis indicated an increase in oxidative phosphorylation in the epidermis of the group-B ( P = 0.039) relative to controls. These findings suggested synergistic in-vivo electroporation-mediated gene transfer as a promising therapeutic approach to enhance viability and vascularity of skin flap. Furthermore, the study showed that combinational gene therapy promoted an increase in tissue perfusion and a relative increase in oxidative metabolism within the epithelium., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Seyed Jafari, Blank, Ramser, Woessner, Shafighi, Geiser, Quinn, Hunger and Gazdhar.)
- Published
- 2021
- Full Text
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246. Enhancing CRISPR deletion via pharmacological delay of DNA-PKcs.
- Author
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Bosch-Guiteras N, Uroda T, Guillen-Ramirez HA, Riedo R, Gazdhar A, Esposito R, Pulido-Quetglas C, Zimmer Y, Medová M, and Johnson R
- Subjects
- Animals, DNA genetics, DNA metabolism, DNA End-Joining Repair, DNA-Activated Protein Kinase metabolism, DNA-Binding Proteins antagonists & inhibitors, DNA-Binding Proteins metabolism, CRISPR-Cas Systems genetics, DNA Breaks, Double-Stranded, DNA-Activated Protein Kinase antagonists & inhibitors, Gene Editing, Sequence Deletion
- Abstract
CRISPR-Cas9 deletion (CRISPR-del) is the leading approach for eliminating DNA from mammalian cells and underpins a variety of genome-editing applications. Target DNA, defined by a pair of double-strand breaks (DSBs), is removed during nonhomologous end-joining (NHEJ). However, the low efficiency of CRISPR-del results in laborious experiments and false-negative results. By using an endogenous reporter system, we show that repression of the DNA-dependent protein kinase catalytic subunit (DNA-PKcs)-an early step in NHEJ-yields substantial increases in DNA deletion. This is observed across diverse cell lines, gene delivery methods, commercial inhibitors, and guide RNAs, including those that otherwise display negligible activity. We further show that DNA-PKcs inhibition can be used to boost the sensitivity of pooled functional screens and detect true-positive hits that would otherwise be overlooked. Thus, delaying the kinetics of NHEJ relative to DSB formation is a simple and effective means of enhancing CRISPR-deletion., (© 2021 Bosch-Guiteras et al.; Published by Cold Spring Harbor Laboratory Press.)
- Published
- 2021
- Full Text
- View/download PDF
247. Non-viral Gene Delivery Methods for Bone and Joints.
- Author
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Gantenbein B, Tang S, Guerrero J, Higuita-Castro N, Salazar-Puerta AI, Croft AS, Gazdhar A, and Purmessur D
- Abstract
Viral carrier transport efficiency of gene delivery is high, depending on the type of vector. However, viral delivery poses significant safety concerns such as inefficient/unpredictable reprogramming outcomes, genomic integration, as well as unwarranted immune responses and toxicity. Thus, non-viral gene delivery methods are more feasible for translation as these allow safer delivery of genes and can modulate gene expression transiently both in vivo , ex vivo , and in vitro . Based on current studies, the efficiency of these technologies appears to be more limited, but they are appealing for clinical translation. This review presents a summary of recent advancements in orthopedics, where primarily bone and joints from the musculoskeletal apparatus were targeted. In connective tissues, which are known to have a poor healing capacity, and have a relatively low cell-density, i.e., articular cartilage, bone, and the intervertebral disk (IVD) several approaches have recently been undertaken. We provide a brief overview of the existing technologies, using nano-spheres/engineered vesicles, lipofection, and in vivo electroporation. Here, delivery for microRNA (miRNA), and silencing RNA (siRNA) and DNA plasmids will be discussed. Recent studies will be summarized that aimed to improve regeneration of these tissues, involving the delivery of bone morphogenic proteins (BMPs), such as BMP2 for improvement of bone healing. For articular cartilage/osteochondral junction, non-viral methods concentrate on targeted delivery to chondrocytes or MSCs for tissue engineering-based approaches. For the IVD, growth factors such as GDF5 or GDF6 or developmental transcription factors such as Brachyury or FOXF1 seem to be of high clinical interest. However, the most efficient method of gene transfer is still elusive, as several preclinical studies have reported many different non-viral methods and clinical translation of these techniques still needs to be validated. Here we discuss the non-viral methods applied for bone and joint and propose methods that can be promising in clinical use., (Copyright © 2020 Gantenbein, Tang, Guerrero, Higuita-Castro, Salazar-Puerta, Croft, Gazdhar and Purmessur.)
- Published
- 2020
- Full Text
- View/download PDF
248. Inhalational delivery of induced pluripotent stem cell secretome improves postpneumonectomy lung structure and function.
- Author
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Dane DM, Cao K, Zhang YA, H Kernstine K, Gazdhar A, Geiser T, and Hsia CCW
- Subjects
- Animals, Dogs, Female, Humans, Lung Volume Measurements, Pulmonary Diffusing Capacity, Induced Pluripotent Stem Cells, Lung physiology, Lung surgery, Pneumonectomy
- Abstract
Cell-free secretory products (secretome) of human induced pluripotent stem cells (iPSCs) have been shown to attenuate tissue injury and facilitate repair and recovery. To examine whether iPSC secretome facilitates mechanically induced compensatory responses following unilateral pneumonectomy (PNX), litter-matched young adult female hounds underwent right PNX (removing 55%-58% of lung units), followed by inhalational delivery of either the nebulized-conditioned media containing induced pluripotent stem cell secretome (iPSC CM) or control cell-free media (CFM); inhalation was repeated every 5 days for 10 treatments. Lung function was measured under anesthesia pre-PNX and 10 days after the last treatment (8 wk post-PNX); detailed quantitative analysis of lung ultrastructure was performed postmortem. Pre-PNX lung function was similar between groups. Compared with CFM control, treatment with iPSC CM attenuated the post-PNX decline in lung diffusing capacity for carbon monoxide and membrane diffusing capacity, accompanied by a 24% larger postmortem lobar volume and distal air spaces. Alveolar double-capillary profiles were 39% more prevalent consistent with enhanced intussusceptive angiogenesis. Frequency distribution of the harmonic mean thickness of alveolar blood-gas barrier shifted toward the lowest values, whereas alveolar septal tissue volume and arithmetic septal thickness were similar, indicating septal remodeling and reduced diffusive resistance of the blood-gas barrier. Thus, repetitive inhalational delivery of iPSC secretome enhanced post-PNX alveolar angiogenesis and septal remodeling that are associated with improved gas exchange compensation. Results highlight the plasticity of the remaining lung units following major loss of lung mass that are responsive to broad-based modulation provided by the iPSC secretome. NEW & NOTEWORTHY To examine whether the secreted products of human induced pluripotent stem cells (iPSCs) facilitate innate adaptive responses following loss of lung tissue, adult dogs underwent surgical removal of one lung, then received repeated administration of iPSC secretory products via inhalational delivery compared with control treatment. Inhalation of iPSC secretory products enhanced capillary formation and beneficial structural remodeling in the remaining lung, leading to improved lung function.
- Published
- 2020
- Full Text
- View/download PDF
249. Transcriptomic profiling reveals disease-specific characteristics of epithelial cells in idiopathic pulmonary fibrosis.
- Author
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Boesch M, Baty F, Brutsche MH, Tamm M, Roux J, Knudsen L, Gazdhar A, Geiser T, Khan P, and Hostettler KE
- Subjects
- Adult, Aged, Cells, Cultured, Epithelial-Mesenchymal Transition genetics, Female, Gene Expression Regulation, Humans, Idiopathic Pulmonary Fibrosis pathology, Lung Diseases, Interstitial, Male, Mesenchymal Stem Cells, Middle Aged, RNA biosynthesis, RNA genetics, Signal Transduction, Epithelial Cells pathology, Gene Expression Profiling, Idiopathic Pulmonary Fibrosis genetics, Transcriptome
- Abstract
Background: Idiopathic pulmonary fibrosis (IPF) is an incurable disease characterized by progressive lung fibrosis ultimately resulting in respiratory failure and death. Recurrent micro-injuries to the alveolar epithelium and aberrant alveolar wound healing with impaired re-epithelialization define the initial steps of the pathogenic trajectory. Failure of timely alveolar epithelial repair triggers hyper-proliferation of mesenchymal cells accompanied by increased deposition of extracellular matrix into the lung interstitium., Methods: We previously isolated fibrosis-specific mesenchymal stem cell (MSC)-like cells from lung tissue of patients with interstitial lung diseases. These cells produced factors bearing anti-fibrotic potential and changed their morphology from mesenchymal to epithelial upon culture in an epithelial cell (EC)-specific growth medium. Here, we set out to molecularly characterize these MSC-like cell-derived ECs using global gene expression profiling by RNA-sequencing. Moreover, we aimed at characterizing disease-specific differences by comparing the transcriptomes of ECs from IPF and non-IPF sources., Results: Our results suggest that differentially expressed genes are enriched for factors related to fibrosis, hypoxia, bacterial colonization and metabolism, thus reflecting many of the hallmark characteristics of pulmonary fibrosis. IPF-ECs showed enrichment of both pro- and anti-fibrotic genes, consistent with the notion of adaptive, compensatory regulation., Conclusions: Our findings support the hypothesis of a functional impairment of IPF-ECs, which could possibly explain the poor clinical outcome of IPF that roughly compares to those of advanced-stage cancers. Our study provides a valuable resource for downstream mechanistic investigation and the quest for novel therapeutic IPF targets.
- Published
- 2020
- Full Text
- View/download PDF
250. Modulation of the unfolded protein response pathway as an antiviral approach in airway epithelial cells.
- Author
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Schögler A, Caliaro O, Brügger M, Oliveira Esteves BI, Nita I, Gazdhar A, Geiser T, and Alves MP
- Subjects
- Bronchi cytology, Bronchi virology, Case-Control Studies, Cells, Cultured, Child, Cystic Fibrosis complications, Endoplasmic Reticulum Chaperone BiP, Epithelial Cells virology, Humans, Respiratory Mucosa cytology, Respiratory Mucosa virology, Rhinovirus physiology, Signal Transduction, Antiviral Agents pharmacology, Endoplasmic Reticulum Stress drug effects, Epithelial Cells drug effects, Rhinovirus drug effects, Unfolded Protein Response, Virus Replication drug effects
- Abstract
Introduction: Rhinovirus (RV) infection is a major cause of cystic fibrosis (CF) lung morbidity with limited therapeutic options. Various diseases involving chronic inflammatory response and infection are associated with endoplasmic reticulum (ER) stress and subsequent activation of the unfolded protein response (UPR), an adaptive response to maintain cellular homeostasis. Recent evidence suggests impaired ER stress response in CF airway epithelial cells, this might be a reason for recurrent viral infection in CF. Therefore, assuming that ER stress inducing drugs have antiviral properties, we evaluated the activation of the UPR by selected ER stress inducers as an approach to control virus replication in the CF bronchial epithelium., Methods: We assessed the levels of UPR markers, namely the glucose-regulated protein 78 (Grp78) and the C/EBP homologous protein (CHOP), in primary CF and control bronchial epithelial cells and in a CF and control bronchial epithelial cell line before and after infection with RV. The cells were also pretreated with ER stress-inducing drugs and RV replication and shedding was measured by quantitative RT-PCR and by a TCID
50 assay, respectively. Cell death was assessed by a lactate dehydrogenate (LDH) activity test in supernatants., Results: We observed a significantly impaired induction of Grp78 and CHOP in CF compare to control cells following RV infection. The ER stress response could be significantly induced in CF cells by pharmacological ER stress inducers Brefeldin A, Tunicamycin, and Thapsigargin. The chemical induction of the UPR pathway prior to RV infection of CF and control cells reduced viral replication and shedding by up to two orders of magnitude and protected cells from RV-induced cell death., Conclusion: RV infection causes an impaired activation of the UPR in CF cells. Rescue of the ER stress response by chemical ER stress inducers reduced significantly RV replication in CF cells. Thus, pharmacological modulation of the UPR might represent a strategy to control respiratory virus replication in the CF bronchial epithelium., (Copyright © 2018 Elsevier B.V. All rights reserved.)- Published
- 2019
- Full Text
- View/download PDF
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