TGF beta inhibits expression of SP-A, SP-B, SP-C, but not SP-D in human alveolar type II cells

Correll, Kelly A.; Edeen, Karen E.; Zemans, Rachel L.; Redente, Elizabeth F.; Mikels‐Vigdal, Amanda; Mason, Robert J.

doi:10.1016/j.bbrc.2018.04.003

Cited by 16 publications

(12 citation statements)

References 43 publications

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“…Expression of SP-C was detectable as a ~ 7 kDa protein band in AT-II cells (4.92 ± 3.98 AU; n = 5) only and varied signi cantly in different cell types (p = 0.001). The sizes of the SP-C and SP-B proteins detected by western blotting were equivalent to sizes previously described [21]. The pattern of β-actin staining differed between non-reduced and reduced samples.…”

Section: Protein Expressionsupporting

confidence: 69%

Primary Nasal Epithelial Cells As a Type-II Alveolar Surrogate Cell Culture Model for ABCA-3 Deficiency

Shaw

Kicic

Fletcher

et al. 2021

Preprint

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ATP Binding Cassette Subfamily A Member 3 (ABCA-3) is a lipid transporter protein highly expressed in type-II alveolar (AT-II) cells. Mutations in ABCA3 can result in severe respiratory disease in infants and children. To study ABCA-3 deficiency in vitro, primary AT-II cells would be the cell culture of choice although sample accessibility is limited. Our aim was to investigate the suitability of primary nasal epithelial cells, as a surrogate culture model for AT-II cells, to study ABCA-3 deficiency. Expression of ABCA3, and surfactant protein genes, SFTPB and SFTPC, was detected in primary nasal epithelial cells but at a significantly lower level than in AT-II cells. ABCA-3, SP-B and SP-C were detected by immunofluorescence microscopy in primary nasal epithelial cells. However, SP-B and SP-C were undetectable in primary nasal epithelial cells using western blot. Structurally imperfect lamellar bodies were observed in primary nasal epithelial cells using transmission electron microscopy. Functional assessment of the ABCA-3 protein demonstrated that higher concentrations of doxorubicin reduced cell viability in ABCA-3 deficient nasal epithelial cells compared to controls in an assay-dependent manner. Our results indicate that there may be a role for primary nasal epithelial cell cultures to model ABCA-3 deficiency in vitro although additional cell culture models that more effectively recapitulate the AT-II phenotype may be required.

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Section: Protein Expressionsupporting

confidence: 69%

Primary Nasal Epithelial Cells As a Type-II Alveolar Surrogate Cell Culture Model for ABCA-3 Deficiency

Shaw

Kicic

Fletcher

et al. 2021

Preprint

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“…Effects of active TGF-β1 on AE2 cells include the downregulation of SPs B and C, and other factors involved in surfactant biosynthesis [11,17]. During the expression of a porcine-derived active TGF-β1, the surface tension lowering capacity of broncho-alveolar lavage-derived surfactant has therefore been shown to be severely impaired.…”

Section: Discussionmentioning

confidence: 99%

“…A previous study showed that severe surfactant dysfunction and microatelectases represent the initial pathological findings during expression of active TGF-β1 and occur in the absence of any fibrotic remodeling in the early phase [11]. Surfactant dysfunction could be attributed to TGF-β1-induced downregulation of biophysically active surfactant proteins (SPs) B and C, and other components involved in surfactant biosynthesis in alveolar epithelial type II (AE2) cells [17], most likely due to lack of activity of thyroid transcription factor-1 in the nucleus [11]. In a second step, an increase in collagen fibrils in interalveolar septal walls was observed on day 14 of this model, whereas AE2 cells were subject to ultrastructural changes, characterized by a loss of surface area of the apical plasma membrane in relation to the baso-lateral plasma membrane [11].…”

Section: Introductionmentioning

confidence: 99%

Surfactant dysfunction and alveolar collapse are linked with fibrotic septal wall remodeling in the TGF-β1-induced mouse model of pulmonary fibrosis

Beike

Wrede

Hegermann

et al. 2019

Laboratory Investigation

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In human idiopathic pulmonary fibrosis (IPF), collapse of distal airspaces occurs in areas of the lung not (yet) remodeled. Mice lungs overexpressing transforming growth factor-β1 (TGF-β1) recapitulate this abnormality: surfactant dysfunction results in alveolar collapse preceding fibrosis and loss of alveolar epithelial type II (AE2) cells' apical membrane surface area. Here we examined whether surfactant dysfunction-related alveolar collapse due to TGF-β1 overexpression is linked to septal wall remodeling and AE2 cell abnormalities. Three and 6 days after gene transfer of TGF-β1, mice received either intratracheal surfactant (Surf-groups: Curosurf®, 100 mg/kg bodyweight) or 0.9% NaCl (Saline-groups). On days 7 (D7) and 14 (D14), lung mechanics were assessed followed by design-based stereology at light and electron microscopic level to quantify structures. Compared with Saline, Surf showed significantly improved tissue elastance, increased numbers of open alveoli, as well as reduced alveolar size heterogeneity on D7. Deterioration in lung mechanics was highly correlated to the loss of open alveoli. On D14, lung mechanics, number of open alveoli, and alveolar size heterogeneity remained significantly improved in the Surf-group. Volumes of extracellular matrix and collagen fibrils in septal walls were significantly reduced, whereas the apical membrane surface area of AE2 cells was increased in Surf compared with Saline. In remodeled tissue with collapsed alveoli, three-dimensional reconstruction of AE2 cells based on scanning electron microscopy array tomography revealed that AE2 cells were trapped without contact to airspaces in the TGF-β1 mouse model. Similar observations were made in human IPF. Based on correlation analyses, the number of open alveoli and of alveolar size heterogeneity were highly linked with the loss of apical membrane surface area of AE2 cells and deposition of collagen fibrils in septal walls on D14. In conclusion, surfactant replacement therapy stabilizes alveoli and prevents extracellular matrix deposition in septal walls in the TGF-β1 model.

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“…Notably, TGF-β1 markedly decreases the expression of fatty acid synthase 42 which we found downregulated in 3G8 cells (−1.95 fold change). This enzyme besides playing a pivotal role in lipid metabolism, is also implicated in the induction of angiogenesis 43 , in particular in cancer cells 44 .…”

Section: Discussionmentioning

confidence: 64%

Runx2 stimulates neoangiogenesis through the Runt domain in melanoma

Cecconi

Brandi

Manfredi

et al. 2019

Sci Rep

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Runx2 is a transcription factor involved in melanoma cell migration and proliferation. Here, we extended the analysis of Runt domain of Runx2 in melanoma cells to deepen understanding of the underlying mechanisms. By the CRISPR/Cas9 system we generated the Runt KO melanoma cells 3G8. Interestingly, the proteome analysis showed a specific protein signature of 3G8 cells related to apoptosis and migration, and pointed out the involvement of Runt domain in the neoangiogenesis process. Among the proteins implicated in angiogenesis we identified fatty acid synthase, chloride intracellular channel protein-4, heat shock protein beta-1, Rho guanine nucleotide exchange factor 1, D-3-phosphoglycerate dehydrogenase, myosin-1c and caveolin-1. Upon querying the TCGA provisional database for melanoma, the genes related to these proteins were found altered in 51.36% of total patients. In addition, VEGF gene expression was reduced in 3G8 as compared to A375 cells; and HUVEC co-cultured with 3G8 cells expressed lower levels of CD105 and CD31 neoangiogenetic markers. Furthermore, the tube formation assay revealed down-regulation of capillary-like structures in HUVEC co-cultured with 3G8 in comparison to those with A375 cells. These findings provide new insight into Runx2 molecular details which can be crucial to possibly propose it as an oncotarget of melanoma.

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TGF beta inhibits expression of SP-A, SP-B, SP-C, but not SP-D in human alveolar type II cells

Cited by 16 publications

References 43 publications

Primary Nasal Epithelial Cells As a Type-II Alveolar Surrogate Cell Culture Model for ABCA-3 Deficiency

Primary Nasal Epithelial Cells As a Type-II Alveolar Surrogate Cell Culture Model for ABCA-3 Deficiency

Surfactant dysfunction and alveolar collapse are linked with fibrotic septal wall remodeling in the TGF-β1-induced mouse model of pulmonary fibrosis

Runx2 stimulates neoangiogenesis through the Runt domain in melanoma

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