Mechanical Stretch Induces Fetal Type II Cell Differentiation Via an Epidermal Growth Factor Receptor–Extracellular-Regulated Protein Kinase Signaling Pathway

Sanchez‐Esteban, Juan; Wang, Yulian; Gruppuso, Philip A.; Rubin, Lewis P.

doi:10.1165/rcmb.2003-0121oc

Cited by 75 publications

(71 citation statements)

References 47 publications

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“…23,24 Briefly, neonatal mouse lungs were first perfused with sterile phosphate-buffered saline and removed en bloc. Collected lungs were dissected, finely minced, and enzymatically digested for a brief period at 37 C in Dulbecco's modified Eagle's medium containing collagenase type I (0.5 mg/mL; SigmaAldrich) and collagenase type IA (0.5 mg/mL; Sigma Aldrich), followed by pipetting to mechanically disperse lung tissue.…”

Section: Mouse Lung Fibroblast Isolation and Culturementioning

confidence: 99%

Epithelial-Derived Inflammation Disrupts Elastin Assembly and Alters Saccular Stage Lung Development

Benjamin

Meer

et al. 2016

The American Journal of Pathology

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The highly orchestrated interactions between the epithelium and mesenchyme required for normal lung development can be disrupted by perinatal inflammation in preterm infants, although the mechanisms are incompletely understood. We used transgenic (inhibitory kB kinase b transactivated) mice that conditionally express an activator of the NF-kB pathway in airway epithelium to investigate the impact of epithelial-derived inflammation during lung development. Epithelial NF-kB activation selectively impaired saccular stage lung development, with a phenotype comprising rapidly progressive distal airspace dilation, impaired gas exchange, and perinatal lethality. Epithelial-derived inflammation resulted in disrupted elastic fiber organization and down-regulation of elastin assembly components, including fibulins 4 and 5, lysyl oxidase likee1, and fibrillin-1. Fibulin-5 expression by saccular stage lung fibroblasts was consistently inhibited by treatment with bronchoalveolar lavage fluid from inhibitory kB kinase b transactivated mice, Escherichia coli lipopolysaccharide, or tracheal aspirates from preterm infants exposed to chorioamnionitis. Expression of a dominant NF-kB inhibitor in fibroblasts restored fibulin-5 expression after lipopolysaccharide treatment, whereas reconstitution of fibulin-5 rescued extracellular elastin assembly by saccular stage lung fibroblasts. Elastin organization was disrupted in saccular stage lungs of preterm infants exposed to systemic inflammation. Our study reveals a critical window for elastin assembly during the saccular stage that is disrupted by inflammatory signaling and could be amenable to interventions that restore elastic fiber assembly in the developing lung.

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Section: Mouse Lung Fibroblast Isolation and Culturementioning

confidence: 99%

Epithelial-Derived Inflammation Disrupts Elastin Assembly and Alters Saccular Stage Lung Development

Benjamin

Meer

et al. 2016

The American Journal of Pathology

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“…Non-adherent cells were collected and cultured overnight. Purity of the FATIIC fraction was determined to be 90 ± 5% by microscopic analysis of epithelial cell morphology and immune-blotting for cytokeratin/ surfactant protein-C and vimentin as markers of epithelial cells and fibroblasts respectively (Sanchez-Esteban et al, 2004). After overnight culture, FATIICs were harvested with trypsin and plated at a density of 10 × 10 5 cells/well on 6-well plates precoated with laminin [10 μg/ml].…”

Section: Cell Isolation Hyperoxia and Treatment Proceduresmentioning

confidence: 99%

Cathepsin B is activated as an executive protease in fetal rat alveolar type II cells exposed to hyperoxia

Lee

Kim

2011

Exp Mol Med

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Alveolar type II cells are main target of hyperoxia-induced lung injury. The authors investigated whether lysosomal protease, cathepsin B (CB), is activated in fetal alveolar type II cells in the transitional period from the canalicular to saccular stages during 65%-hyperoxia and whether CB is related to fetal alveolar type II cell (FATIIC) death secondary to hyperoxia. FATIICs were isolated from embryonic day 19 rats and exposed to 65%-oxygen for 24 h and 36 h. The cells exposed to room air were used as controls. Cell cytotoxicity was assessed by lactate dehydrogenase-release and flow cytometry, and apoptosis was analyzed by TUNEL assay and flow cytometry. CB activity was assessed by colorimetric assay, qRT-PCR and western blots. 65%-hyperoxia induced FATIIC death via necrosis and apoptosis. Interestingly, caspase-3 activities were not enhanced in FATIICs during 65%-hyperoxia, whereas CB activities were greatly increased during 65%-hyperoxia in a time-dependent manner, and similar findings were observed with qRT-PCR and western blots. In addition, the preincubation of CB inhibitor prior to 65%-hyperoxia reduced FATIIC death significantly. Our studies suggest that CB activation secondary to hyperoxia might have a relevant role in executing the cell death program in FATIICs during the acute stage of 65%-hyperoxia.

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“…Epithelial cells were then seeded onto Bioflex plates precoated with collagen-1 (Flexcell Corporation, Hillsborough, NC) and maintained in serum-free Dulbecco's modified Eagle's medium (DMEM) for 24 h. Plates containing adherent cells were mounted in a Flexercell FX-4000 Strain Unit (Flexcell Corp.). Equibiaxial elongation of 5% was applied at intervals of 60 cycles per minute for 15 min plus 2.5% continuous distention for the remaining 45 min of each hour, for 16 h. This regimen was chosen to simulate mechanical forces experienced by type II epithelial cells during lung development (13), and it is based on previous studies from our laboratory (5,8). Cells grown on nonstrained substrates were treated in an identical manner and served as controls.…”

Section: Methodsmentioning

confidence: 99%

DNA Microarray Reveals Novel Genes Induced by Mechanical Forces in Fetal Lung Type II Epithelial Cells

et al. 2006

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Mechanical forces are essential for normal fetal lung development. However, the cellular and molecular mechanisms regulating this process are still poorly defined. In this study, we used oligonucleotide microarrays to investigate gene expression in cultured embryonic d 19 rat fetal lung type II epithelial cells exposed to a level of mechanical strain similar to the developing lung. Significance Analysis of Microarrays (SAM) identified 92 genes differentially expressed by strain. Interestingly, several members of the solute carrier family of amino acid transporter (Slc7a1, Slc7a3, Slc6a9, and tumor-associated protein 1) genes involved in amino acid synthesis (Phgdh, Psat1, Psph, Cars, and Asns), as well as the amiloride-sensitive epithelial sodium channel gene (Scnn1a) were up-regulated by the application of force. These results were confirmed by quantitative real-time PCR (qRT-PCR). Thus, this study identifies genes induced by strain that may be important for amino acid signaling pathways and protein synthesis in fetal type II cells. In addition, these data suggest that mechanical forces may contribute to facilitate lung fluid reabsorption in preparation for birth. Taken together, the present investigation provides further insights into how mechanical forces may modulate fetal lung development. N ormal lung growth and development during fetal life are critical for extrauterine survival. Premature infants are often born before sufficient lung maturation has occurred, and, as a result, they experience a high rate of long-term pulmonary complications such as bronchopulmonary dysplasia. Understanding the mechanisms of prenatal lung growth and development is a crucial step for the design of preventive and therapeutic strategies.Mechanical forces generated in utero by repetitive breathing movements and by fluid distension are essential to mammalian lung development (1-3). Previous in vitro experiments have demonstrated that application of force to cultured type II epithelial cells induces cell proliferation (4) and differentiation (5). Other studies have begun to identify mechanoreceptors and cell signaling pathways mediating fetal lung growth and maturation (4 -8). However, the precise molecular and cellular mechanisms by which lung cells sense mechanical stimuli to influence lung development are still poorly defined.DNA microarray technology provides a powerful tool for rapid, comprehensive, and quantitative analysis of gene expression profile. This technique has been previously used to assess changes in gene expression in endothelial cells (9), bladder smooth muscle cells (10), and human pulmonary A549 (11) and H441 (12) epithelial cells exposed to mechanical strain.Therefore, the goal of the present study was to identify genes differentially expressed by mechanical stress of fetal type II epithelial cells. We used an in vitro model system in which embryonic d (E) 19 type II cells are exposed to mechanical forces similar to that observed in the developing lung. These studies revealed that mechanical force...

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Mechanical Stretch Induces Fetal Type II Cell Differentiation Via an Epidermal Growth Factor Receptor–Extracellular-Regulated Protein Kinase Signaling Pathway

Cited by 75 publications

References 47 publications

Epithelial-Derived Inflammation Disrupts Elastin Assembly and Alters Saccular Stage Lung Development

Epithelial-Derived Inflammation Disrupts Elastin Assembly and Alters Saccular Stage Lung Development

Cathepsin B is activated as an executive protease in fetal rat alveolar type II cells exposed to hyperoxia

DNA Microarray Reveals Novel Genes Induced by Mechanical Forces in Fetal Lung Type II Epithelial Cells

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