Bleomycin and its Role in Inducing Apoptosis and Senescence in Alveolar Epithelial Lung Cells - Modulating Effects of Caveolin-1: An Update

In this study an in vitro exposure test to investigate toxicological effects of the volatile disinfection by-product trichloramine and of real indoor pool air was established. For this purpose a set-up to generate a well-defined, clean gas stream of trichloramine was combined with biotests. Human alveolar epithelial lung cells of the cell line A-549 were exposed in a CULTEX® device with trichloramine concentrations between 0.1 and 40 mg/m3 for 1 h. As toxicological endpoints the cell viability and the inflammatory response by the cytokines IL-6 and IL-8 were investigated. A decreasing cell viability could be observed with increasing trichloramine concentration. An increase of IL-8 release could be determined at trichloramine concentrations higher than 10 mg/m3 and an increase of IL-6 release at concentrations of 20 mg/m3. Investigations of indoor swimming pool air showed similar inflammatory effects to the lung cells although the air concentrations of trichloramine of 0.17 and 0.19 mg/m3 were much lower compared with the laboratory experiments with trichloramine as the only contaminant. Therefore it is assumed that a mixture of trichloramine and other disinfection by-products in the air of indoor pool settings contribute to that effect.

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Immortalisation of primary human alveolar epithelial lung cells using a non-viral vector to study respiratory bioreactivity in vitro

Scientific Reports ◽

10.1038/s41598-020-77191-y ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Alberto Katsumiti ◽

Pakatip Ruenraroengsak ◽

Miren P. Cajaraville ◽

Andrew J. Thorley ◽

Teresa D. Tetley

Keyword(s):

Viral Vector ◽

Alveolar Epithelium ◽

Simian Virus 40 ◽

Differential Sensitivity ◽

Molecular Characteristics ◽

Lung Cells ◽

Alveolar Epithelial ◽

Alkaline Phosphatase Staining ◽

Epithelial Lung Cells

AbstractTo overcome the scarcity of primary human alveolar epithelial cells for lung research, and the limitations of current cell lines to recapitulate the phenotype, functional and molecular characteristics of the healthy human alveolar epithelium, we have developed a new method to immortalise primary human alveolar epithelial lung cells using a non-viral vector to transfect the telomerase catalytic subunit (hTERT) and the simian virus 40 large-tumour antigen (SV40). Twelve strains of immortalised cells (ICs) were generated and characterised using molecular, immunochemical and morphological techniques. Cell proliferation and sensitivity to polystyrene nanoparticles (PS) were evaluated. ICs expressed caveolin-1, podoplanin and receptor for advanced glycation end-products (RAGE), and most cells were negative for alkaline phosphatase staining, indicating characteristics of AT1-like cells. However, most strains also contained some cells that expressed pro-surfactant protein C, classically described to be expressed only by AT2 cells. Thus, the ICs mimic the cellular heterogeneity in the human alveolar epithelium. These ICs can be passaged, replicate rapidly and remain confluent beyond 15 days. ICs showed differential sensitivity to positive and negatively charged PS nanoparticles, illustrating their potential value as an in vitro model to study respiratory bioreactivity. These novel ICs offer a unique resource to study human alveolar epithelial biology.

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TRIM72 is required for effective repair of alveolar epithelial cell wounding

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00172.2014 ◽

2014 ◽

Vol 307 (6) ◽

pp. L449-L459 ◽

Cited By ~ 15

Author(s):

Seong Chul Kim ◽

Thomas Kellett ◽

Shaohua Wang ◽

Miyuki Nishi ◽

Nagaraja Nagre ◽

...

Keyword(s):

Epithelial Cells ◽

Molecular Mechanisms ◽

Striated Muscle ◽

Alveolar Epithelial Cell ◽

Alveolar Epithelial Cells ◽

Lung Cells ◽

Alveolar Epithelial ◽

High Tidal Volume Ventilation

The molecular mechanisms for lung cell repair are largely unknown. Previous studies identified tripartite motif protein 72 (TRIM72) from striated muscle and linked its function to tissue repair. In this study, we characterized TRIM72 expression in lung tissues and investigated the role of TRIM72 in repair of alveolar epithelial cells. In vivo injury of lung cells was introduced by high tidal volume ventilation, and repair-defective cells were labeled with postinjury administration of propidium iodide. Primary alveolar epithelial cells were isolated and membrane wounding and repair were labeled separately. Our results show that absence of TRIM72 increases susceptibility to deformation-induced lung injury whereas TRIM72 overexpression is protective. In vitro cell wounding assay revealed that TRIM72 protects alveolar epithelial cells through promoting repair rather than increasing resistance to injury. The repair function of TRIM72 in lung cells is further linked to caveolin 1. These data suggest an essential role for TRIM72 in repair of alveolar epithelial cells under plasma membrane stress failure.

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Caveolin-1 Expression and Caveolae Biogenesis during Cell Transdifferentiation in Lung Alveolar Epithelial Primary Cultures

Biochemical and Biophysical Research Communications ◽

10.1006/bbrc.1999.1280 ◽

1999 ◽

Vol 262 (3) ◽

pp. 744-751 ◽

Cited By ~ 88

Author(s):

Lee Campbell ◽

Andrew J. Hollins ◽

Ahmed Al-Eid ◽

Geoff R. Newman ◽

Chris von Ruhland ◽

...

Keyword(s):

Primary Cultures ◽

Caveolin 1 ◽

Alveolar Epithelial ◽

Cell Transdifferentiation

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Podoplanin interaction with caveolin-1 promotes tumour cell migration and invasion

10.1101/488304 ◽

2018 ◽

Cited By ~ 1

Author(s):

Luisa Pedro ◽

Jacqueline D. Shields

Keyword(s):

Tumour Cell ◽

Surface Expression ◽

Alveolar Epithelial Cells ◽

Migration And Invasion ◽

Dependent Manner ◽

Caveolin 1 ◽

Alveolar Epithelial ◽

Podoplanin Expression

AbstractPodoplanin, a highly O-glycosylated type-1 transmembrane glycoprotein, found in lymphatic endothelial cells, podocytes, alveolar epithelial cells and lymph node fibroblasts is also expressed by tumour cells, and is correlated with more aggressive disease. Despite numerous studies documenting podoplanin expression, the mechanisms underlying its tumour-promoting functions remain unclear. Using a murine melanoma cell line that endogenously expresses podoplanin, we demonstrate interactions with proteins necessary for cytoskeleton reorganization, adhesion and matrix degradation, and endocytosis/receptor recycling but also identify a novel interaction with caveolin-1. We generated a panel of podoplanin and caveolin-1 variants to determine the molecular interactions and functional consequences of these interactions. Complementary in vitro and in vivo systems confirmed the existence of a functional cooperation in which surface expression of both full length, signalling competent podoplanin and caveolin-1 are necessary to induce directional migration and invasion, which is executed via PAK1 and ERK1 pathways. Our findings establish that podoplanin signalling mediates the invasive properties of melanoma cells in a caveolin-1 dependent manner.Summary StatementThis manuscript describes a new interaction and functional cooperation between podoplanin and caveolin1 that drives tumour cell invasion into surrounding tissues.

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Paracrine Regulation of Alveolar Epithelial Damage and Repair Responses by Human Lung-Resident Mesenchymal Stromal Cells

Cells ◽

10.3390/cells10112860 ◽

2021 ◽

Vol 10 (11) ◽

pp. 2860

Author(s):

Dennis M. L. W. Kruk ◽

Marissa Wisman ◽

Jacobien A. Noordhoek ◽

Mehmet Nizamoglu ◽

Marnix R. Jonker ◽

...

Keyword(s):

Epithelial Cells ◽

Mesenchymal Stromal Cells ◽

Lung Tissue ◽

Stromal Cells ◽

Wound Repair ◽

Lung Cells ◽

Alveolar Type ◽

Type Ii ◽

Epithelial Damage ◽

Alveolar Epithelial

COPD is characterized by irreversible lung tissue damage. We hypothesized that lung-derived mesenchymal stromal cells (LMSCs) reduce alveolar epithelial damage via paracrine processes, and may thus be suitable for cell-based strategies in COPD. We aimed to assess whether COPD-derived LMSCs display abnormalities. LMSCs were isolated from lung tissue of severe COPD patients and non-COPD controls. Effects of LMSC conditioned-medium (CM) on H2O2-induced, electric field- and scratch-injury were studied in A549 and NCI-H441 epithelial cells. In organoid models, LMSCs were co-cultured with NCI-H441 or primary lung cells. Organoid number, size and expression of alveolar type II markers were assessed. Pre-treatment with LMSC-CM significantly attenuated oxidative stress-induced necrosis and accelerated wound repair in A549. Co-culture with LMSCs supported organoid formation in NCI-H441 and primary epithelial cells, resulting in significantly larger organoids with lower type II-marker positivity in the presence of COPD-derived versus control LMSCs. Similar abnormalities developed in organoids from COPD compared to control-derived lung cells, with significantly larger organoids. Collectively, this indicates that LMSCs’ secretome attenuates alveolar epithelial injury and supports epithelial repair. Additionally, LMSCs promote generation of alveolar organoids, with abnormalities in the supportive effects of COPD-derived LMCS, reflective of impaired regenerative responses of COPD distal lung cells.

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