TY  - JOUR
AB  - The development of chronic lung disease in the neonate, also known as bronchopulmonary dysplasia (BPD), is the most common long-term complication in prematurely born infants. In BPD, the disease-characteristic inflammatory response culminates in nonreversible remodeling of the developing gas exchange area, provoked by the impact of postnatal treatments such as mechanical ventilation (MV) and oxygen treatment. To evaluate the potential of prenatal treatment regimens to modulate this inflammatory response and thereby impact the vulnerability of the lung toward postnatal injury, we designed a multilayered preclinical mouse model. After administration of either prenatal vitamin D-enriched (VitD+; 1,500 IU/g food) or -deprived (VitD-; <10 IU/kg) food during gestation in C57B6 mice (the onset of mating until birth), neonatal mice were exposed to hyperoxia (FiO2 = 0.4) with or without MV for 8 h at days 5-7 of life, whereas controls spontaneously breathed room air. Prenatal vitamin D supplementation resulted in a decreased number of monocytes/macrophages in the neonatal lung undergoing postnatal injury together with reduced TGF-β pathway activation. In consequence, neonatal mice that received a VitD+ diet during gestation demonstrated less extracellular matrix (ECM) remodeling upon lung injury, reflected by the reduction of pulmonary α-smooth muscle actin-positive fibroblasts, decreased collagen and elastin deposition, and lower amounts of interstitial tissue in the lung periphery. In conclusion, our findings support strategies that attempt to prevent vitamin D insufficiency during pregnancy as they could impact lung health in the offspring by mitigating inflammatory changes in neonatal lung injury and ameliorating subsequent remodeling of the developing gas exchange area.NEW & NOTEWORTHY Vitamin D-enriched diet during gestation resulted in reduced lung inflammation and matrix remodeling in neonatal mice exposed to clinically relevant, postnatal injury. The results underscore the need to monitor the subclinical effects of vitamin D insufficiency that impact health in the offspring when other risk factors come into play.
AU  - Heydarian, M.
AU  - Oak, P.
AU  - Koschlig, M.
AU  - Kamgari, N.
AU  - Hagemann, M.
AU  - Wjst, M.*
AU  - Hilgendorff, A.
C1  - 67867
C2  - 54345
CY  - 6120 Executive Blvd, Suite 600, Rockville, Md, United States
SP  - L95-L103
TI  - Prenatal vitamin D supplementation mitigates inflammation-related alveolar remodeling in neonatal mice.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 325
IS  - 2
PB  - Amer Physiological Soc
PY  - 2023
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - OBJECTIVE: Neonatal chronic lung disease lacks standardised assessment of lung structural changes. METHOD AND RESULTS: We addressed this clinical need by the development of a novel scoring system (UNSEAL BPD (UNiforme Scoring of the disEAsed Lung in BPD)) using T2-weighted single-shot fast-spin-echo sequences from 3T MRI in very premature infants with and without bronchopulmonary dysplasia (BPD). Quantification of interstitial and airway remodeling, emphysematous changes and ventilation inhomogeneity was achieved by consensus scoring on a 5-point Likert scale. We successfully identified moderate and severe disease by logistic regression (AUC 0.89) complemented by classification tree analysis revealing gestational age-specific structural changes. We demonstrated substantial inter-reader reproducibility (weighted Cohen's kappa 0.69) and disease specificity (AUC=0.91). CONCLUSION: Our novel MRI score enables the standardised assessment of disease characteristic structural changes in the preterm lung exhibiting significant potential as a quantifiable endpoint in early intervention clinical trials and long-term disease monitoring.
AU  - Förster, K.
AU  - Marchi, H.
AU  - Stöcklein, S.*
AU  - Dietrich, O.*
AU  - Ehrhardt, H.*
AU  - Wielpütz, M.O.*
AU  - Flemmer, A.W.*
AU  - Schubert, B.
AU  - Mall, M.A.*
AU  - Ertl-Wagner, B.*
AU  - Hilgendorff, A.
C1  - 66864
C2  - 53335
SP  - L114-L122
TI  - MRI based scoring of the diseased lung in the preterm infant with BPD.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 324
IS  - 2
PY  - 2022
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - The 9th biennial conference titled "Stem Cells, Cell Therapies, and Bioengineering in Lung Biology and Diseases" was hosted virtually, due to the ongoing COVID-19 pandemic, in collaboration with the University of Vermont Larner College of Medicine, the National Heart, Lung, and Blood Institute, the Alpha-1 Foundation, the Cystic Fibrosis Foundation, and the International Society for Cell & Gene Therapy. The event was held from July 12th through 15th, 2021 with a pre-conference workshop held July 9th. As in previous years the objectives remained to review and discuss the status of active research areas involving stem cells, cellular therapeutics, and bioengineering as they relate to the human lung. Topics included: 1) technological advancements in the in situ analysis of lung tissues, 2) new insights into stem cell signalling and plasticity in lung remodelling and regeneration, 3) the impact of extracellular matrix in stem cell regulation and airway engineering in lung regeneration, 4) differentiating and delivering stem cell therapeutics to the lung, 5) regeneration in response to viral infection, and 6) ethical development of cell-based treatments for lung diseases. This selection of topics represents some of the most dynamic and current research areas in lung biology. The virtual workshop included active discussion on state-of-the-art methods relating to the core features of the 2021 conference, including in-situ protemics, lung-on-chip, iPSC-airway differentiation, and light sheet microscopy. The conference concluded with an open discussion to suggest funding priorities and recommendations for future research directions in basic and translational lung biology.
AU  - Ikonomou, L.*
AU  - Magnusson, M.*
AU  - Dries, R.*
AU  - Herzog, E.L.*
AU  - Hynds, R.*
AU  - Borok, Z.*
AU  - Park, J.A.*
AU  - Skolasinski, S.*
AU  - Burgess, J.K.*
AU  - Turner, L.*
AU  - Mojarad, S.M.*
AU  - Mahoney, J.E.*
AU  - Lynch, T.*
AU  - Lehmann, M.
AU  - Thannickal, V.J.*
AU  - Hook, J.L.*
AU  - Vaughan, A.E.*
AU  - Hoffman, E.T.*
AU  - Weiss, D.J.*
AU  - Ryan, A.L.*
C1  - 65514
C2  - 52708
SP  - L341-L354
TI  - Stem cells, cell therapies, and bioengineering in lung biology and disease 2021.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 323
IS  - 3
PY  - 2022
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - RATIONALE: The bronchial epithelium is constantly challenged by inhalative insults including cigarette smoke (CS), a key risk factor for lung disease. In vitro exposure of bronchial epithelial cells using CS extract (CSE) is a widespread alternative to whole CS (wCS) exposure. However, CSE exposure protocols vary considerably between studies, precluding direct comparison of applied doses. Moreover, they are rarely validated in terms of physiological response in vivo and the relevance of the findings is often unclear. METHODS: We tested six different exposure settings in primary human bronchial epithelial cells (phBECs), including five CSE protocols in comparison with wCS exposure. We quantified cell-delivered dose and directly compared all exposures using expression analysis of 10 well-established smoke-induced genes in bronchial epithelial cells. CSE exposure of phBECs was varied in terms of differentiation state, exposure route, duration of exposure, and dose. Gene expression was assessed by quantitative Real-Time PCR (qPCR) and Western Blot analysis. Cell type-specific expression of smoke-induced genes was analyzed by immunofluorescent analysis. RESULTS: Three surprisingly dissimilar exposure types, namely chronic CSE treatment of differentiating phBECs, acute CSE treatment of submerged basal phBECs, and wCS exposure of differentiated phBECs performed best, resulting in significant upregulation of seven (chronic CSE) and six (acute wCS, acute submerged CSE exposure) out of 10 genes. Acute apical or basolateral exposure of differentiated phBECs with CSE was much less effective despite similar doses used. CONCLUSIONS: Our findings provide guidance for the design of human in vitro CS exposure models in experimental and translational lung research.
AU  - Mastalerz, M.
AU  - Dick, E.
AU  - Chakraborty, A.
AU  - Hennen, E.
AU  - Schamberger, A.C.
AU  - Schröppel, A.
AU  - Lindner, M.*
AU  - Hatz, R.A.*
AU  - Behr, J.*
AU  - Hilgendorff, A.
AU  - Schmid, O.
AU  - Staab-Weijnitz, C.A.
C1  - 63460
C2  - 51541
CY  - 6120 Executive Blvd, Suite 600, Rockville, Md, United States
SP  - L129-L148
TI  - Validation of in vitro models for smoke exposure of primary human bronchial epithelial cells.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 322
IS  - 1
PB  - Amer Physiological Soc
PY  - 2021
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Cigarette smoking has marked effects on lung tissue, including induction of oxidative stress, inflammatory cell recruitment, and a protease/antiprotease imbalance. These effects contribute to tissue remodeling and destruction resulting in loss of lung function in chronic obstructive pulmonary disease (COPD) patients. Cathepsin S (CatS) is a cysteine protease that is involved in the remodeling/degradation of connective tissue and basement membrane. Aberrant expression or activity of CatS has been implicated in a variety of diseases, including arthritis, cancer, cardiovascular, and lung diseases. However, little is known about the effect of cigarette smoking on both CatS expression and activity, as well as its role in smoking-related lung diseases. Here, we evaluated the expression and activity of human CatS in lung tissues from never-smokers and smokers with or without COPD. Despite the presence of an oxidizing environment, CatS expression and activity were significantly higher in current smokers (both non-COPD and COPD) compared with never-smokers, and correlated positively with smoking history. Moreover, we found that the exposure of primary human bronchial epithelial cells to cigarette smoke extract triggered the activation of P2X7 receptors, which in turns drives CatS upregulation. The present data suggest that excessive CatS expression and activity contribute, beside other proteases, to the deleterious effects of cigarette smoke on pulmonary homeostasis.
AU  - Andrault, P.-M.*
AU  - Schamberger, A.C.
AU  - Chazeirat, T.*
AU  - Sizaret, D.*
AU  - Renault, J.*
AU  - Staab-Weijnitz, C.A.
AU  - Hennen, E.
AU  - Petit-Courty, A.*
AU  - Wartenberg, M.*
AU  - Saidi, A.*
AU  - Baranek, T.*
AU  - Guyetant, S.*
AU  - Courty, Y.*
AU  - Eickelberg, O.
AU  - Lalmanach, G.*
AU  - Lecaille, F.*
C1  - 57294
C2  - 47668
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L625-L638
TI  - Cigarette smoke induces overexpression of active human cathepsin S in lungs from current smokers with or without COPD.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 317
IS  - 5
PB  - Amer Physiological Soc
PY  - 2019
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Transforming growth factor-beta (TGF-beta)-induced fibroblast-to-myofibroblast differentiation contributes to remodeling in chronic obstructive pulmonary disease and idiopathic pulmonary fibrosis, but whether this impacts the ability of fibroblasts to support lung epithelial repair remains little explored. We pretreated human lung fibroblasts [primary (phFB) or MRC5 cells] with recombinant human TGF-beta to induce myofibroblast differentiation, then cocultured them with adult mouse lung epithelial cell adhesion molecule-positive cells (EpCAM(+)) to investigate their capacity to support epithelial organoid formation in vitro. While control phFB and MRC5 lung fibroblasts supported organoid formation of mouse EpCAM(+) cells, TGF-beta pretreatment of both phFB and MRC5 impaired organoid-supporting ability. We performed RNA sequencing of TGF beta-treated phFB. which revealed altered expression of key Wnt signaling pathway components and Wnt/beta-catenin target genes, and modulated expression of secreted factors involved in mesenchymal-epithelial signaling. TGF-beta profoundly skewed the transcriptional program induced by the Wnt/beta-catenin activator CHIR99021. Supplementing organoid culture media recombinant hepatocyte growth factor or fibroblast growth factor 7 promoted organoid formation when using TGF-beta pretreated fibroblasts. In conclusion, TGF-beta-induced myofibroblast differentiation results in Wnt/beta-catenin pathway skewing and impairs fibroblast ability to support epithelial repair likely through multiple mechanisms, including modulation of secreted growth factors.
AU  - Ng-Blichfeldt, J.P.
AU  - de Jong, T.*
AU  - Kortekaas, R.K.*
AU  - Wu, X.*
AU  - Lindner, M.
AU  - Guryev, V.*
AU  - Hiemstra, P.S.*
AU  - Stolk, J.*
AU  - Königshoff, M.
AU  - Gosens, R.*
C1  - 56591
C2  - 47108
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L14-L28
TI  - TGF-beta activation impairs fibroblast ability to support adult lung epithelial progenitor cell organoid formation.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 317
IS  - 1
PB  - Amer Physiological Soc
PY  - 2019
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Chronic obstructive pulmonary disease (COPD) is a life-threatening lung disease. Although cigarette smoke was considered the main cause of development, the heterogeneous nature of the disease leaves it unclear whether other factors contribute to the predisposition or impaired regeneration response observed. Recently, epigenetic modification has emerged to be a key player in the pathogenesis of COPD. The addition of methyl groups to arginine residues in both histone and nonhistone proteins by protein arginine methyltransferases (PRMTs) is an important posttranslational epigenetic modification event regulating cellular proliferation, differentiation, apoptosis, and senescence. Here, we hypothesize that coactivator-associated arginine methyltransferase-1 (CARM1) regulates airway epithelial cell injury in COPD pathogenesis by controlling cellular senescence. Using the naphthalene (NA)-induced mouse model of airway epithelial damage, we demonstrate that loss of CC10-positive club cells is accompanied by a reduction in CARM1-expressing cells of the airway epithelium. Furthermore, Carm1 haploinsuffficent mice showed perturbed club cell regeneration following NA treatment. In addition, CARM1 reduction led to decreased numbers of antisenescent sirtuin 1-expressing cells accompanied by higher p21, p16, and beta-galactosidase-positive senescent cells in the mouse airway following NA treatment. Importantly, CARM1-silenced human bronchial epithelial cells showed impaired wound healing and higher beta-galactosidase activity. These results demonstrate that CARM1 contributes to airway repair and regeneration by regulating airway epithelial cell senescence.
AU  - Sarker, R.S.
AU  - Conlon, T.M.
AU  - Morrone, C.
AU  - Srivastava, B.
AU  - Konyalilar, N.*
AU  - Verleden, S.E.*
AU  - Bayram, H.*
AU  - Fehrenbach, H.*
AU  - Yildirim, A.Ö.
C1  - 57315
C2  - 47702
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L602-L614
TI  - CARM1 regulates senescence during airway epithelial cell injury in COPD pathogenesis.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 317
IS  - 5
PB  - Amer Physiological Soc
PY  - 2019
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Asthma is characterized by —Asthma a chronic inflammation and remodeling of the airways. Although inflammation can be controlled, therapeutic options to revert remodeling do not exist. Thus, there is a large and unmet need to understand the underlying molecular mechanisms to develop novel therapies. We previously identified a pivotal role for miR-142-3p in regulating airway smooth muscle (ASM) precursor cell proliferation during lung development by fine-tuning the Wingless/Integrase I (WNT) signaling. Thus, we here aimed to investigate the relevance of this interaction in asthma. We performed quantitative RT-PCR and immune staining in a murine model for ovalbumin-induced allergic airway inflammation and in bronchial biopsies from patients with asthma and isolated primary fibroblasts thereof. miR-142-3p was increased in hyperproliferative regions of lung in murine and human asthma, whereas this microRNA (miRNA) was excluded from regions with differentiated ASM cells. Increases in miR-142-3p were associated with a decrease of its known target Adenomatous polyposis coli. Furthermore, we observed a differential expression of miR-142-3p in bronchial biopsies from patients with early or late onset severe asthma, which coincided with a differential WNT signature. Our data suggest that miR-142-3p is involved in regulating the balance between proliferation and differentiation of ASM cells in asthma, possibly via controlling WNT signaling. Thus, this miRNA might be an interesting target to prevent ASM hyperproliferation in asthma.
AU  - Bartel, S.*
AU  - Carraro, G.*
AU  - Alessandrini, F.
AU  - Krauss-Etschmann, S.*
AU  - Ricciardolo, F.L.M.*
AU  - Bellusci, S.*
C1  - 53476
C2  - 44879
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L328-L333
TI  - MiR-142-3p is associated with aberrant WNT signaling during airway remodeling in asthma.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 315
IS  - 2
PB  - Amer Physiological Soc
PY  - 2018
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Cues from the extracellular matrix (ECM) and their functional interplay with cells play pivotal roles for development, tissue repair, and disease. However, the precise nature of this interplay remains elusive. We used an innovative 3D cell culture ECM model by decellularizing 300-µm-thick ex vivo lung tissue scaffolds (d3D-LTCs) derived from diseased and healthy mouse lungs, which widely mimics the native (patho)physiological in vivo ECM microenvironment. We successfully repopulated all d3D-LTCs with primary human and murine fibroblasts, and moreover, we demonstrated that the cells also populated the innermost core regions of the d3D-LTCs in a real 3D fashion. The engrafted fibroblasts revealed a striking functional plasticity, depending on their localization in distinct ECM niches of the d3D-LTCs, affecting the cells' tissue engraftment, cellular migration rates, cell morphologies, and protein expression and phosphorylation levels. Surprisingly, we also observed fibroblasts that were homing to the lung scaffold's interstitium as well as fibroblasts that were invading fibrotic areas. To date, the functional nature and even the existence of 3D cell matrix adhesions in vivo as well as in 3D culture models is still unclear and controversial. Here, we show that attachment of fibroblasts to the d3D-LTCs evidently occurred via focal adhesions, thus advocating for a relevant functional role in vivo. Furthermore, we found that protein levels of talin, paxillin, and zyxin and phosphorylation levels of paxillin Y118, as well as the migration-relevant small GTPases RhoA, Rac, and CDC42, were significantly reduced compared with their attachment to 2D plastic dishes. In summary, our results strikingly indicate that inherent physical or compositional characteristics of the ECM act as instructive cues altering the functional behavior of engrafted cells. Thus, d3D-LTCs might aid to obtain more realistic data in vitro, with a high relevance for drug discovery and mechanistic studies alike.
AU  - Burgstaller, G.
AU  - Sengupta, A.
AU  - Vierkotten, S.
AU  - Preissler, G.*
AU  - Lindner, M.*
AU  - Behr, J.*
AU  - Königshoff, M.
AU  - Eickelberg, O.*
C1  - 52749
C2  - 44312
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L708-L723
TI  - Distinct niches within the extracellular matrix dictate fibroblast function in (cell free) 3D lung tissue cultures.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 314
IS  - 5
PB  - Amer Physiological Soc
PY  - 2018
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Fibroblasts play an important role in lung homeostasis and disease. In lung fibrosis, fibroblasts adopt a proliferative and migratory phenotype, with increased expression of alpha-smooth muscle actin (alpha SMA) and enhanced secretion of extracellular matrix components. Comprehensive profiling of fibroblast heterogeneity is limited because of a lack of specific cell-surface markers. We have previously profiled the surface proteome of primary human lung fibroblasts. Here, we sought to define and quantify a panel of cluster of differentiation (CD) markers in primary human lung fibroblasts and idiopathic pulmonary fibrosis (IPF) lung tissue, using immunofluorescence and FACS analysis. Fibroblast function was assessed by analysis of replicative senescence. We observed the presence of distinct fibroblast phenotypes in vivo, characterized by various combinations of Desmin, alpha SMA, CD36, or CD97 expression. Most markers demonstrated stable expression over passages in vitro, but significant changes were observed for CD36, CD54, CD82, CD106, and CD140a. Replicative senescence of fibroblasts was observed from passage 10 onward. CD36- and CD97-positive but alpha SMA-negative cells were present in remodeled areas of IPF lungs. Transforming growth factor (TGF)-beta treatment induced alpha SMA and collagen I expression but repressed CD36 and CD97 expression. We identified a panel of stable surface markers in human lung fibroblasts, applicable for positive-cell isolation directly from lung tissue. TGF-beta exposure represses CD36 and CD97 expression, despite increasing alpha SMA expression; we therefore identified complex surface protein changes during fibroblast-myofibroblast activation. Coexistence of quiescence and activated fibroblast subtypes in the IPF lung suggests dynamic remodeling of fibroblast activation upon subtle changes to growth factor exposure in local microenvironmental niches.
AU  - Heinzelmann, K.
AU  - Lehmann, M.
AU  - Gerckens, M.
AU  - Noskovicova, N.
AU  - Frankenberger, M.
AU  - Lindner, M.
AU  - Hatz, R.*
AU  - Behr, J.*
AU  - Hilgendorff, A.
AU  - Königshoff, M.
AU  - Eickelberg, O.
C1  - 53729
C2  - 44990
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L682-L696
TI  - Cell-surface phenotyping identifies CD36 and CD97 as novel markers of fibroblast quiescence in lung fibrosis.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 315
IS  - 5
PB  - Amer Physiological Soc
PY  - 2018
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Fibroblasts are thought to be the prime cell type for producing and secreting extracellular matrix (ECM) proteins in the connective tissue. The profibrotic cytokine transforming growth factor-β1 (TGF-β1) activates and transdifferentiates fibroblasts into α-smooth muscle actin (α-SMA)-expressing myofibroblasts, which exhibit increased ECM secretion, in particular collagens. Little information, however, exists about cell-surface molecules on fibroblasts that mediate this transdifferentiation process. We recently identified, using unbiased cell-surface proteome analysis, Cub domain-containing protein 1 (CDCP1) to be strongly downregulated by TGF-β1. CDCP1 is a transmembrane glycoprotein, the expression and role of which has not been investigated in lung fibroblasts to date. Here, we characterized, in detail, the effect of TGF-β1 on CDCP1 expression and function, using immunofluorescence, FACS, immunoblotting, and siRNA-mediated knockdown of CDCP1. CDCP1 is present on interstitial fibroblasts, but not myofibroblasts, in the normal and idiopathic pulmonary fibrosis lung. In vitro, TGF-β1 decreased CDCP1 expression in a time-dependent manner by impacting mRNA and protein levels. Knockdown of CDCP1 enhanced a TGF-β1-mediated cell adhesion of fibroblasts. Importantly, CDCP1-depleted cells displayed an enhanced expression of profibrotic markers, such as collagen V or α-SMA, which was found to be independent of TGF-β1. Our data show, for the very first time that loss of CDCP1 contributes to fibroblast to myofibroblast differentiation via a potential negative feedback loop between CDCP1 expression and TGF-β1 stimulation.
AU  - Noskovicova, N.
AU  - Heinzelmann, K.
AU  - Burgstaller, G.
AU  - Behr, J.*
AU  - Eickelberg, O.
C1  - 52753
C2  - 44480
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L695-L707
TI  - Cub domain-containing protein 1 negatively regulates TGF-β signaling and myofibroblast differentiation.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 314
IS  - 5
PB  - Amer Physiological Soc
PY  - 2018
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Mesenchymal stromal cells (MSCs) are released into the airways of preterm infants following lung injury. These cells display a proinflammatory phenotype and are associated with development of severe bronchopulmonary dysplasia (BPD). We aimed to characterize the functional properties of MSCs obtained from tracheal aspirates of 50 preterm infants who required invasive ventilation. Samples were separated by disease severity. The increased proliferative capacity of MSCs was associated with longer duration of mechanical ventilation and higher severity of BPD. Augmented growth depended on nuclear accumulation of NF kappa Bp65 and was accompanied by reduced expression of cytosolic alpha-smooth muscle actin (alpha-SMA). The central role of NF-kappa B signaling was confirmed by inhibition of I kappa B alpha phosphorylation. The combined score of proliferative capacity, accumulation of NF kappa Bp65, and expression of alpha-SMA was used to predict the development of severe BPD with an area under the curve (AUC) of 0.847. We mimicked the clinical situation in vitro, and stimulated MSCs with IL-1 beta and TNF-alpha. Both cytokines induced similar and persistent changes as was observed in MSCs obtained from preterm infants with severe BPD. RNA interference was employed to investigate the mechanistic link between NF kappa Bp65 accumulation and alterations in phenotype. Our data indicate that determining the phenotype of resident pulmonary MSCs represents a promising biomarker-based approach. The persistent alterations in phenotype, observed in MSCs from preterm infants with severe BPD, were induced by the pulmonary inflammatory response. NF kappa Bp65 accumulation was identified as a central regulatory mechanism. Future preclinical and clinical studies, aimed to prevent BPD, should focus on phenotype changes in pulmonary MSCs.
AU  - Reicherzer, T.
AU  - Häffner, S.
AU  - Shahzad, T.*
AU  - Gronbach, J.*
AU  - Mysliwietz, J.
AU  - Huebener, C.*
AU  - Hasbargen, U.*
AU  - Gertheiss, J.*
AU  - Schulze, A.*
AU  - Bellusci, S.*
AU  - Morty, R.E.*
AU  - Hilgendorff, A.
AU  - Ehrhardt, H.*
C1  - 53396
C2  - 44846
CY  - 9650 Rockville Pike, Bethesda, Md 20814 Usa
SP  - L87-L101
TI  - Activation of the NF-kappa B pathway alters the phenotype of MSCs in the tracheal aspirates of preterm infants with severe BPD.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 315
IS  - 1
PB  - Amer Physiological Soc
PY  - 2018
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - IPF is a devastating chronic interstitial lung disease (ILD) characterized by lung tissue scarring and high morbidity. Lung epithelial injury, myofibroblast activation, and deranged repair are believed to be key processes involved in disease onset and progression but the exact molecular mechanisms behind IPF remain unclear. Several drugs have been shown to slow disease progression, but treatments which halt or reverse IPF progression have not been identified. Ex vivo models of human lung have been proposed for drug discovery, one of which is precision-cut lung slices (PCLS). Although PCLS production from IPF explants is possible, IPF explants are rare and typically represent end-stage disease. Here we present a novel model of early fibrosis-like changes in human PCLS derived from patients without ILD/IPF using a combination of profibrotic growth factors and signaling molecules. Fibrotic-like changes of PCLS were qualitatively analyzed by histology and immunofluorescence and quantitatively by WST1, RT-qPCR, WB, and ELISA. PCLS remained viable after 5 days of treatment and fibrotic gene expression (FN1, SERPINE1, COL1A1, CTGF, MMP7 and ACTA2) increased as early as 24h of treatment, with increases in protein levels at 48 hours and increased deposition of extracellular matrix. Alveolar epithelium reprogramming was evident by decreases in SFTPC and loss of HOPX In summary, using human-derived PCLS from patients without ILD/IPF, we established a novel ex vivo model which displays characteristics of early fibrosis and could be used to evaluate novel therapies and study early-stage IPF pathomechanisms.
AU  - Alsafadi, H.N.
AU  - Staab-Weijnitz, C.A.
AU  - Lehmann, M.
AU  - Lindner, M.*
AU  - Peschel, B.
AU  - Königshoff, M.
AU  - Wagner, D.E.
C1  - 50755
C2  - 42512
CY  - Bethesda
SP  - L896-L902
TI  - An ex vivo model to induce early fibrosis-like changes in human precision-cut lung slices.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 312
IS  - 6
PB  - Amer Physiological Soc
PY  - 2017
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - RATIONALE: Bronchopulmonary dysplasia (BPD), characterized by impaired alveolarization and vascularization in association with lung inflammation and apoptosis, often occurs after mechanical ventilation with oxygen rich gas (MV-O2). As heightened expression of the pro-inflammatory cytokine TNF-α has been described in infants with BPD, we hypothesized that absence of TNF-α would reduce pulmonary inflammation, and attenuate structural changes in newborn mice undergoing MV-O2 Methods: Neonatal TNF-α null (TNF-α(-/-)) and wild type (TNF-α(+/+)) mice received MV-O2for 8h; controls spontaneously breathed 40%O2. Histologic, mRNA and protein analysis in vivo were complemented by in vitro studies subjecting primary pulmonary myofibroblasts to mechanical stretch. Finally, TNF-α level in tracheal aspirates (TA) from preterm infants were determined by ELISA. RESULTS: Although MV-O2induced larger and fewer alveoli in both, TNF-α(-/-)and TNF-α(+/+)mice, it caused enhanced lung apoptosis (TUNEL, Caspase-3/-6/-8), infiltration of macrophages and neutrophils, and pro-inflammatory mediator expression (IL-1β, CXCL-1, MCP-1) in TNF-α(-/-)mice. These differences were associated with increased pulmonary TGF-β signaling, decreased TGF-β inhibitor SMAD-7 expression and reduced pulmonary NF-κB activity in ventilated TNF-α(-/-)mice. Preterm infants who went on to develop BPD showed significantly lower TNF-α levels at birth. CONCLUSION: Our results suggest a critical balance between TNF-α and TGF-β signaling in the developing lung, and underscore the critical importance of these key pathways in the pathogenesis of BPD. Future treatment strategies need to weigh the potential benefits of inhibiting pathologic cytokine expression against the potential of altering key developmental pathways.
AU  - Ehrhardt, H.*
AU  - Pritzke, T.
AU  - Oak, P.
AU  - Kossert, M.
AU  - Biebach, L.*
AU  - Forster, K.*
AU  - Koschlig, M.
AU  - Alvira, C.M.*
AU  - Hilgendorff, A.
C1  - 48197
C2  - 41051
CY  - Bethesda
SP  - L909-L918
TI  - Absence of TNF-α enhances inflammatory response in the newborn lung undergoing mechanical ventilation.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 310
IS  - 10
PB  - Amer Physiological Soc
PY  - 2016
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - To date, phenotyping and disease course prediction in idiopathic pulmonary fibrosis (IPF) primarily relies on lung function measures. Blood biomarkers were recently proposed for diagnostic and outcome prediction in IPF, yet their correlation with lung function and histology remains unclear. Here, we comprehensively assessed biomarkers in liquid biopsies and correlated their abundance with lung function and histology during the onset, progression, and resolution of lung fibrosis, with the aim to more precisely evaluate disease progression in the pre-clinical model of bleomycin-induced pulmonary fibrosis in vivo. Importantly, the strongest correlation of lung function with histological extent of fibrosis was observed at day 14, while lung function was unchanged at day 28 and 56, even when histology showed marked fibrotic lesions. While MMP7, MMP9, and PAI1 were significantly elevated in BAL of fibrotic mice, only sICAM1 was elevated in the peripheral blood of fibrotic mice and strongly correlated with the extent of fibrosis. Importantly, tissue-bound ICAM1 was also elevated in lung homogenates, with prominent staining in hyperplastic type II alveolar epithelial and endothelial cells. In sum, we show that lung function decline is not a prerequisite for histologically evident fibrosis, particularly during the onset or resolution thereof. Plasma levels of sICAM1 strongly correlate with the extent of lung fibrosis, and may thus be considered for the assessment of intraindividual therapeutic studies in preclinical studies of pulmonary fibrosis.
AU  - Fernandez, I.E.
AU  - Amarie, O.V.
AU  - Mutze, K.
AU  - Königshoff, M.
AU  - Yildirim, A.Ö.
AU  - Eickelberg, O.
C1  - 48160
C2  - 39948
CY  - Bethesda
SP  - 919-927
TI  - Systematic phenotyping and correlation of biomarkers with lung function and histology in lung fibrosis.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 310
IS  - 10
PB  - Amer Physiological Soc
PY  - 2016
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Epidemiological evidence demonstrates a strong link between postnatal cigarette smoke (CS)-exposure and increased respiratory morbidity in young children. However, how CS induces early onset airways disease in young children and how it interacts with endogenous risk factors remains poorly understood. We, therefore exposed 10 day old neonatal wild-type and βENaC-transgenic mice with cystic fibrosis like lung disease to CS for 4 days. Neonatal wild-type mice exposed to CS demonstrated increased numbers of macrophages and neutrophils in the BALF which was accompanied by increased levels of Mmp12 and Cxcl1. BALF from βENaC-transgenic mice contained greater numbers of macrophages which did not increase following acute CS-exposure, however there was significant increase in airway neutrophilia compared to filtered air transgenic and CS-exposed wild-type controls. Interestingly, wild-type and βENaC-transgenic mice demonstrated epithelial airway and vascular remodeling following CS-exposure. Morphometric analysis of lung sections revealed that CS-exposure caused increased mucus accumulation in the airway lumen of neonatal βENaC-transgenic mice compared to wild-type controls, which was accompanied by an increase in the number of goblet cells and Muc5ac upregulation. We conclude that short-term CS exposure i) induces acute airways disease with airway epithelial and vascular remodeling in neonatal wild-type mice; and ii) exacerbates airway inflammation, mucus hypersecretion and mucus plugging in neonatal βENaC-transgenic mice with chronic lung disease. Our results in neonatal mice suggest that young children may be highly susceptible to develop airways disease in response to tobacco smoke exposure and that adverse effects may be aggravated in children with underlying chronic lung diseases.
AU  - Jia, J.
AU  - Conlon, T.M.
AU  - Seimetz, M.*
AU  - Bednorz, M.*
AU  - Zhou-Suckow, Z.*
AU  - Weissmann, N.*
AU  - Eickelberg, O.
AU  - Mall, M.A.*
AU  - Yildirim, A.Ö.
C1  - 49173
C2  - 41708
CY  - Bethesda
SP  - L602-L610
TI  - Cigarette smoke causes acute airways disease and exacerbates chronic obstructive lung disease in neonatal mice.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 311
IS  - 3
PB  - Amer Physiological Soc
PY  - 2016
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - The proteasome system degrades more than 80% of intracellular proteins into small peptides. Accordingly, the proteasome is involved in many essential cellular functions such as protein quality control, transcription, immune responses, cell signaling, and apoptosis. Moreover, degradation products are loaded onto major histocompatibility (MHC) class I molecules to communicate the intracellular protein composition to the immune system. The standard 20S proteasome core complex contains three distinct catalytic active sites that are exchanged upon stimulation with inflammatory cytokines to form the so-called immunoproteasome. Immunoproteasomes are constitutively expressed in immune cells and have different proteolytic activities compared to standard proteasomes. They are rapidly induced in parenchymal cells upon intracellular pathogen infection and are crucial for priming effective CD8+ T cell-mediated immune responses against infected cells. Beyond shaping these adaptive immune reactions, immunoproteasomes also regulate the function of immune cells by degradation of inflammatory and immune mediators. Accordingly, they emerge as novel regulators of innate immune responses. The recently unraveled impairment of immunoproteasome function by environmental challenges and by genetic variations of immunoproteasome genes might represent a currently underestimated risk factor for the development and progression of lung diseases. In particular, immunoproteasome dysfunction will dampen resolution of infections thereby promoting exacerbations, may foster autoimmunity in chronic lung diseases, and possibly contributes to immune evasion of tumor cells. Novel pharmacological tools such as site-specific inhibitors of the immunoproteasome as well as activity-based probes, however, hold promises as novel therapeutic drugs for respiratory diseases and biomarker profiling, respectively.
AU  - Kammerl, I.E.
AU  - Meiners, S.
C1  - 48907
C2  - 41471
CY  - Bethesda
SP  - L328-L336
TI  - Proteasome function shapes innate and adaptive immune responses.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 311
IS  - 2
PB  - Amer Physiological Soc
PY  - 2016
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - During the last decades, the study of cell behavior was largely accomplished in uncoated or extracellular matrix (ECM)-coated plastic dishes. To date, considerable cell biological efforts have tried to model in vitro the natural microenvironment found in vivo. For the lung, explants cultured ex vivo as lung tissue cultures (LTCs) provide a 3D tissue model containing all cells in their natural microenvironment. Techniques for assessing the dynamic live interaction between ECM and cellular tissue components, however, are still missing. Here, we describe specific multidimensional immunolabeling of living 3D-LTCs, derived from healthy and fibrotic mouse lungs, as well as patient-derived 3D-LTCs, and concomitant real-time 4D multi-channel imaging thereof. This approach allowed the evaluation of dynamic interactions between mesenchymal cells and macrophages with their ECM. Furthermore, fibroblasts transiently expressing focal adhesions markers incorporated into the 3D-LTCs, paving new ways for studying the dynamic interaction between cellular adhesions and their natural-derived ECM. A novel protein transfer technology (FuseIt/Ibidi) shuttled fluorescently-labelled αSMA antibodies into the native cells of living 3D-LTCs, enabling live monitoring of αSMA-positive stress fibers in native tissue myofibroblasts residing in fibrotic lesions of 3D-LTCs. Finally, this technique can be applied to healthy and diseased human lung tissue, as well as to adherent cells in conventional 2D cell culture. This novel method will provide valuable new insights into the dynamics of ECM (patho)biology, studying in detail the interaction between ECM and cellular tissue components in their natural microenvironment.
AU  - Burgstaller, G.
AU  - Vierkotten, S.
AU  - Lindner, M.*
AU  - Königshoff, M.
AU  - Eickelberg, O.
C1  - 45363
C2  - 37301
CY  - Bethesda
SP  - L323-L332
TI  - Multidimensional immunolabeling and 4D time-lapse imaging of vital ex vivo lung tissue.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 309
IS  - 4
PB  - Amer Physiological Soc
PY  - 2015
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Elastin plays a pivotal role in lung development. We therefore queried if elastin haplo-insufficient newborn mice (Eln(+/-)) would exhibit abnormal lung structure and function related to modified extracellular matrix (ECM) composition. As mechanical ventilation (MV) has been linked to dysregulated elastic fiber formation in the newborn lung, we also asked if elastin haplo-insufficiency would accentuate lung growth arrest seen after prolonged MV of neonatal mice. We studied 5d-old wild-type (Eln(+/+)) and Eln(+/-) littermates at baseline, and after MV with air for 8-24h. Lungs of unventilated Eln(+/-) mice contained ~50% less elastin and ~100% more collagen-1 and lysyl oxidase compared to Eln(+/+) pups. Eln(+/-) lungs contained fewer capillaries than Eln(+/+) lungs, without discernible differences in alveolar structure. In response to MV, lung tropoelastin and elastase activity increased in Eln(+/+) neonates, whereas tropoelastin decreased and elastase activity was unchanged in Eln(+/-) mice. Fibrillin-1 protein increased in lungs of both groups during MV, more in Eln(+/-) than in Eln(+/+) pups. In both groups, MV caused capillary loss, with larger and fewer alveoli compared with unventilated controls. Respiratory system elastance, which was less in unventilated Eln(+/-) compared to Eln(+/+) mice, was similar in both groups after MV. These results suggest that elastin haplo-insufficiency adversely impacts pulmonary angiogenesis, and that MV dysregulates elastic fiber integrity, with further loss of lung capillaries, lung growth arrest, and impaired respiratory function in both Eln(+/+) and Eln(+/-) mice. Paucity of lung capillaries in Eln(+/-) newborns might help explain subsequent development of pulmonary hypertension previously reported in adult Eln(+/-) mice.
AU  - Hilgendorff, A.
AU  - Parai, K.*
AU  - Ertsey, R.*
AU  - Navarro, E.F.*
AU  - Jain, N.*
AU  - Carandang, F.*
AU  - Peterson, J.*
AU  - Mokres, L.M.*
AU  - Milla, C.*
AU  - Preuss, S.*
AU  - Alejandre Alcazar, M.A.*
AU  - Khan, S.*
AU  - Masumi, J.*
AU  - Ferreira-Tojais, N.*
AU  - Mujahid, S.*
AU  - Starcher, B.C.*
AU  - Rabinovitch, M.*
AU  - Bland, R.D.*
C1  - 43032
C2  - 35980
CY  - Bethesda
SP  - L464-L478
TI  - Lung matrix and vascular remodeling in mechanically ventilated elastin haplo-insufficient (Eln+/-) newborn mice.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 308
IS  - 5
PB  - Amer Physiological Soc
PY  - 2015
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Microfibrillar-associated protein 4 (MFAP4) is localized to elastic fibers in blood vessels and the interalveolar septa of the lungs and is further present in bronchoalveolar lavage. Mfap4 has been previously suggested to be involved in elastogenesis in the lung. We tested this prediction and aimed to characterize the pulmonary function changes and emphysematous changes that occur in Mfap4 deficient (Mfap4-/-) mice. Significant changes included increases in total lung capacity and compliance, which were evident in Mfap4-/- mice at 6 months and 8 months, but not at 3 months of age. Using in vivo breath-hold gated micro-computed tomography (micro-CT) in 8-month-old Mfap4-/- mice, we found that the mean density of the lung parenchyma was decreased, and the low-attenuation area (LAA) was significantly increased by 14 % compared to Mfap4+/+ mice. Transmission electron microscopy (TEM) did not reveal differences in the organization of elastic fibers, and there was no difference in elastin content, but borderline significant increase in elastin mRNA expression in 3-month-old mice. Stereological analysis showed that alveolar surface density in relation to the lung parenchyma and total alveolar surface area inside of the lung were both significantly decreased in Mfap4-/- mice by 25 % and 15 %, respectively. The data did not support an essential role of MFAP4 in pulmonary elastic fiber organization or content, but indicated increased turnover in young Mfap4-/- mice. However, Mfap4-/- mice developed a spontaneous loss of lung function, which was evident at 6 months of age, and moderate airspace enlargement, with emphysema-like changes.
AU  - Holm, A.T.*
AU  - Wulf-Johansson, H.*
AU  - Hvidtsen, S.*
AU  - Jorgensen, P.T.*
AU  - Schlosser, A.*
AU  - Pilecki, B.*
AU  - Ormhoj, M.*
AU  - Moeller, J.B.*
AU  - Johannsen, C.*
AU  - Baun, C.*
AU  - Andersen, T.*
AU  - Schneider, J.P.*
AU  - Hegermann, J.*
AU  - Ochs, M.*
AU  - Götz, A.A.*
AU  - Schulz, H.
AU  - Hrabě de Angelis, M.
AU  - Vestbo, J.*
AU  - Holmskov, U.*
AU  - Sorensen, G.L.*
C1  - 44439
C2  - 36832
CY  - Bethesda
SP  - L1114-L1124
TI  - Characterization of spontaneous air space enlargement in mice lacking microfibrillar-associated protein 4.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 308
IS  - 11
PB  - Amer Physiological Soc
PY  - 2015
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Fibroblast growth factors (Fgfs) mediate organ repair. Lung epithelial cell overexpression of Fgf10 post-bleomycin injury is both protective and therapeutic, characterized by increased survival and attenuated fibrosis. Exogenous administration of FGF7 (Palifermin) also showed prophylactic survival benefits in mice. The role of endogenous Fgfr2b ligands upon bleomycin-induced lung fibrosis is still elusive. This study reports the expression of endogenous Fgfr2b ligands, receptors and signaling targets in wild type mice following bleomycin lung injury. In addition, the impact of attenuating endogenous Fgfr2b-ligands following bleomycin-induced fibrosis was tested using a doxycycline (dox)-based inducible, soluble, dominant-negative form of the Fgfr2b receptor. Double transgenic (DTG) Rosa26(rtTA/+);tet(O)solFgfr2b mice were validated for the expression and activity of soluble Fgfr2b (failure to regenerate maxillary incisors, attenuated recombinant FGF7 signal in the lung). As previously reported, no defects in lung morphometry were detected in DTG (+dox) mice exposed from post-natal (PN) 1 through PN105. Female single transgenic (STG) and DTG mice were subjected to various levels of bleomycin injury (1.0, 2.0, and 3.0U/kg). Fgfr2b ligands were attenuated either throughout injury (d0-d11; d0-28) or during later stages (d6-d28 and d14-d28). No significant changes in survival, weight, lung function, confluent areas of fibrosis, or hydroxyproline deposition were detected in DTG mice. These results indicate that endogenous Fgfr2b ligands do not significantly protect against bleomycin injury, nor do they expedite the resolution of bleomycin lung induced injury in mice.
AU  - MacKenzie, B.*
AU  - Henneke, I.*
AU  - Hezel, S.*
AU  - Al Alam, D.*
AU  - El Agha, E.*
AU  - Chao, C.M.*
AU  - Quantius, J.*
AU  - Wilhelm, J.*
AU  - Jones, M.*
AU  - Goth, K.*
AU  - Li, X.*
AU  - Seeger, W.*
AU  - Königshoff, M.
AU  - Herold, S.*
AU  - Rizvanov, A.A.*
AU  - Günther, A.*
AU  - Bellusci, S.*
C1  - 44076
C2  - 36789
CY  - Bethesda
SP  - L1014-L1024
TI  - Attenuating endogenous Fgfr2b ligands during bleomycin-induced lung fibrosis does not compromise murine lung repair.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 308
IS  - 10
PB  - Amer Physiological Soc
PY  - 2015
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Cigarette smoke is the main risk factor for chronic obstructive pulmonary disease (COPD). Exposure of cells to cigarette smoke induces an initial adaptive cellular stress response involving increased oxidative stress and induction of inflammatory signaling pathways. Exposure of mitochondria to cellular stress alters their fusion/fission dynamics. While mild stress induces a pro-survival response termed stress induced mitochondrial hyperfusion, severe stress results in mitochondrial fragmentation and mitophagy. In the present study, we analyzed the mitochondrial response to mild and non-toxic doses of cigarette smoke extract (CSE) in alveolar epithelial cells. We characterized mitochondrial morphology, expression of mitochondrial fusion and fission genes, markers of mitochondrial proteostasis as well as mitochondrial functions such as membrane potential and oxygen consumption. Murine lung epithelial (MLE)12, as well as primary mouse alveolar epithelial cells revealed pronounced mitochondrial hyperfusion upon treatment with CSE, accompanied by increased expression of the mitochondrial fusion protein mitofusin (MFN) 2 and increased metabolic activity. We did not observe any alterations in mitochondrial proteostasis, i.e. induction of the mitochondrial unfolded protein response or mitophagy. Therefore, our data indicate an adaptive pro-survival response of mitochondria of alveolar epithelial cells to non-toxic concentrations of CSE. A hyperfused mitochondrial network, however, renders the cell more vulnerable to additional stress such as sustained cigarette smoke exposure. As such cigarette smoke induced mitochondrial hyperfusion - although being part of a beneficial adaptive stress response in the first place - may contribute to the pathogenesis of COPD.
AU  - Ballweg, K.
AU  - Mutze, K.
AU  - Königshoff, M.
AU  - Eickelberg, O.
AU  - Meiners, S.
C1  - 32571
C2  - 35147
SP  - 895-907
TI  - Cigarette smoke extract affects mitochondrial function in alveolar epithelial cells.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 307
IS  - 11
PY  - 2014
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Copyright © 2014 the American Physiological Society. Hemeoxygenase-1 (HO-1), an inducible heat shock protein, is upregulated in response to multiple cellular insults via oxidative stress, lipopolysaccharides (LPS), and hypoxia. In this study, we investigated in vitro the role of Toll-like receptor 4 (TLR4), hypoxia-inducible factor 1α (HIF-1α), and iron on HO-1 expression in cystic fibrosis (CF). Immunohistochemical analysis of TLR4, HO-1, ferritin, and HIF-1α were performed on lung sections of CFTR-/- and wild-type mice. CFBE41o- and 16HBE14o- cell lines were employed for in vitro analysis via immunoblotting, immunofluorescence, real-time PCR, luciferase reporter gene analysis, and iron quantification. We observed a reduced TLR4, HIF-1α, HO-1, and ferritin in CFBE41o- cell line and CF mice. Knockdown studies using TLR4-siRNA in 16HBE14o- revealed significant decrease of HO-1, confirming the role of TLR4 in HO-1 downregulation. Inhibition of HO-1 using tin protoporphyrin in 16HBE14o- cells resulted in increased iron levels, suggesting a probable role of HO-1 in iron accumulation. Additionally, sequestration of excess iron using iron chelators resulted in increased hypoxia response element response in CFBE41o- and 16HBE14o-, implicating a role of iron in HIF-1α stabilization and HO-1. To conclude, our in vitro results demonstrate that multiple regulatory factors, such as impaired TLR4 surface expression, increased intracellular iron, and decreased HIF-1α, downregulate HO-1 expression in CFBE41o- cells.
AU  - Chillappagari, S.*
AU  - Venkatesan, S.*
AU  - Garapati, V.*
AU  - Mahavadi, P.*
AU  - Munder, A.*
AU  - Seubert, A.*
AU  - Sarode, G.*
AU  - Guenther, A.*
AU  - Schmeck, B.T.*
AU  - Tuemmler, B.*
AU  - Henke, M.O.
C1  - 42996
C2  - 35941
SP  - L791-L799
TI  - Impaired TLR4 and HIF expression in cystic fibrosis bronchial epithelial cells downregulates hemeoxygenase-1 and alters iron homeostasis in vitro.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 307
IS  - 10
PY  - 2014
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - Chronic obstructive pulmonary disease (COPD) is characterized by a progressive decline in lung function, caused by exposure to exogenous particles, mainly cigarette smoke (CS). COPD is initiated and perpetuated by an abnormal CS-induced inflammatory response of the lungs, involving both innate and adaptive immunity. Specifically, B cells organized in iBALT structures and macrophages accumulate in the lungs and contribute to CS-induced emphysema, but the mechanisms thereof remain unclear. Here, we demonstrate that B cell-deficient mice are significantly protected against CS-induced emphysema. Chronic CS exposure led to an increased size and number of iBALT structures, and increased lung compliance and mean linear chord length in WT, but not B cell-deficient mice. The increased accumulation of lung resident macrophages around iBALT and in emphysematous alveolar areas in CS-exposed WT mice coincided with upregulated MMP12 expression. In vitro co-culture experiments using B cells and macrophages demonstrated that B cell-derived IL-10 drives macrophage activation and MMP12 upregulation, which could be inhibited by an anti-IL10 antibody. In summary, B cell function in iBALT formation seems necessary for macrophage activation and tissue destruction in CS-induced emphysema, and possibly provides a new target for therapeutic intervention in COPD.
AU  - John-Schuster, G.
AU  - Hager, K.
AU  - Conlon, T.M.
AU  - Irmler, M.
AU  - Beckers, J.
AU  - Eickelberg, O.
AU  - Yildirim, A.Ö.
C1  - 31947
C2  - 34885
CY  - Bethesda
SP  - L692-L706
TI  - Cigarette smoke-induced iBALT mediates macrophage activation in a B cell-dependent manner in COPD.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 307
IS  - 9
PB  - Amer Physiological Soc
PY  - 2014
SN  - 1040-0605
ER  - 

TY  - JOUR
AB  - van Rijt SH, Keller IE, John G, Kohse K, Yildirim AO, Eickelberg O, Meiners S. Acute cigarette smoke exposure impairs proteasome function in the lung. Am J Physiol Lung Cell Mol Physiol 303: L814-L823, 2012. First published September 7, 2012; doi:10.1152/ajplung.00128.2012.-Cigarette smoke mediates DNA damage, lipid peroxidation, and modification and misfolding of proteins, thereby inducing severe cellular damage. The ubiquitin proteasome system serves as the major disposal system for modified and misfolded proteins and is thus essential for proper cellular function. Its role in cigarette smoke-induced cell damage, however, is largely unknown. We hypothesized that the ubiquitin-proteasome system is involved in the degradation of cigarette smoke-damaged proteins and that cigarette smoke exposure impairs the proteasome itself. Here, we show that treatment of human alveolar epithelial cells with cigarette smoke extract (CSE) induced time-and dose-dependent cell death, a rise in intracellular reactive oxygen species, and increased levels of carbonylated and polyubiquitinated proteins. While high doses of CSE severely impaired all three proteasomal activities, low CSE concentrations significantly inhibited only the trypsin-like activity of the proteasome in alveolar and bronchial epithelial cells. Moreover, acute exposure of mice to cigarette smoke significantly impaired the trypsin-like activity by 25% in the lungs. Reduced proteasome activity was not due to transcriptional regulation of the proteasome. Notably, cigarette smoke exposure induced accumulation of polyubiquitinated proteins in the soluble and insoluble protein fraction of the lung. We show for the first time that acute exposure to cigarette smoke directly impairs proteasome activity in the lungs of mice and in human epithelial cells at low doses without affecting proteasome expression. Our results indicate that defective proteasomal protein quality control may exacerbate the detrimental effects of cigarette smoke in the lung.
AU  - van Rijt, S.H.
AU  - Keller, I.E.
AU  - John, G.
AU  - Kohse, K.
AU  - Yildirim, A.Ö.
AU  - Eickelberg, O.
AU  - Meiners, S.
C1  - 11103
C2  - 30522
SP  - L814-L823
TI  - Acute cigarette smoke exposure impairs proteasome function in the lung.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 303
IS  - 9
PB  - Amer. Physiological Soc.
PY  - 2012
SN  - 1040-0605
ER  - 

TY  - JOUR
AU  - Kim, Y.-M.*
AU  - Reed, W.*
AU  - Lenz, A.-G.
AU  - Jaspers, I.*
AU  - Silbajoris, R.*
AU  - Nick, H.S.*
AU  - Samet, J.M.*
C1  - 5072
C2  - 22711
SP  - 432-441
TI  - Ultrafine carbon particles induce interleukin-8 gene transcription and p38 MAPK activation in normal human bronchial epithelial cells.
JO  - Am. J. Physiol. Lung Cell Mol. Physiol.
VL  - 288
PY  - 2005
SN  - 1040-0605
ER  -