We investigated whether in vitro expansion of human alveolar epithelial type II cells is possible. We found that human endogenous human alveolar epithelial type II cells can be cultured and passaged. The culture system enabled retroviral gene transduction into human alveolar epithelial type II cells. We performed RNA sequencing of human alveolar epithelial type II cells transduced with mutant surfactant protein C or control vector. Overall design: Cultured human alveolar epithelial type II cells were transfected with retroviral vector containing mutant surfactant protein C or control retroviral vector. The retroviral vector contained LNGFR as a marker. After gene transduction, transduced cells were purified by magnetic-activated cell sorting. The transcriptome of the cells was generated by 5'Tag-seq using Ion Genestudio S5 Sequencer.
In vitro expansion of endogenous human alveolar epithelial type II cells in fibroblast-free spheroid culture.
Specimen part, Subject
View SamplesIn the alveoli, lung fibroblasts are in close contact with alveolar epithelial cells type 2, and are considered to support alveolar epithelial cells, forming an alveolar stem cell niche. However, what fibroblast-to-epithelial cell interactions occur during the alveolar maturation stage remains unclear. To understand the lung fibroblast-to-epithelial cell interactions, we performed time-course 3´SAGE-seq analysis of lung epithelial cells and fibroblasts. Overall design: Lung epithelial cells and lung fibroblasts from various developmental stages (E18.5, P0.5, P2, P7, P28, and P56) were purified by cell sorting. The time series transcriptome of the epithelial cells and fibroblasts was generated by 3'SAGE-seq using Ion Proton sequencer.
Mesenchymal-Epithelial Interactome Analysis Reveals Essential Factors Required for Fibroblast-Free Alveolosphere Formation.
Specimen part, Cell line, Subject
View SamplesIntratracheal transfer of isolated lung fibroblasts in bleomycin-induced lung fibrosis recapitulates the activation process of lung fibroblasts after epithelial injury. In order to investigate gene expression signatures of transferred fibroblasts, we purified transferred fibroblasts 2, 4, and 7 days after the transfer and performed transcriptome analysis. We also isolated Acta2 high and low cells by using Acta2-mKO1 reporter mice 4 days after the transfer. Overall design: Lung fibroblasts were isolated from untreated Col-GFP mice after tissue dissociation and negative selection for lineage markers. Isolated lung fibroblasts were intratracheally transferred into wild type mice, which received intratracheal bleomycin treatment 7 days before the transfer. Col-GFP+ cells were purified from the host lungs by FACS sorting on 2, 4, and 7 days after the transfer. Acta2 high and low cells were prepared by transferring lung fibroblasts from Acta2-mKO1 reporter mice. mRNA was isolated from sorted cells, and gene expression profiles were acquired by next generation sequencing.
Gli signaling pathway modulates fibroblast activation and facilitates scar formation in pulmonary fibrosis.
Cell line, Subject
View SamplesTumor budding is a definite prognostic marker, but does not always show favorable reproducibility.
Clinical Significance of a Gene Signature Generated from Tumor Budding Grade in Colon Cancer.
Specimen part
View SamplesQuantitative phosphoproteome and transcriptome analysis of ligand-stimulated MCF-7 human breast cancer cells was performed to understand the mechanisms of tamoxifen resistance at a systems level. Phosphoproteome data revealed that wild type (WT) cells were more enriched with phospho-proteins than tamoxifen-resistant (TamR) cells after stimulation with ligands. Surprisingly, decreased phosphorylation after ligand perturbation was more common than increased phosphorylation. In particular, 17beta-estradiol (E2) induced down-regulation in WT cells at a very high rate. E2 and the ErbB ligand, heregulin (HRG) induced almost equal numbers of up-regulated phospho-proteins in WT cells. Pathway and motif activity analyses using transcriptome data additionally suggested that deregulated activation of GSK3B(glycogen synthase kinase 3 beta) and MAPK1/3 signaling might be associated with altered activation of CREB and AP-1 transcription factors in TamR cells and this hypothesis was validated by reporter assays. An examination of clinical samples revealed that, inhibitory phosphorylation of GSK3B at serine 9 was significantly lower in tamoxifen-treated breast cancer patients that eventually had relapses, implying that activation of GSK3B may be associated with the tamoxifen resistant phenotype. Thus, the combined phosphoproteome and transcriptome dataset analyses revealed distinct signal-transcription programs in tumor cells and provided a novel molecular target to understand tamoxifen resistance.
Integrated quantitative analysis of the phosphoproteome and transcriptome in tamoxifen-resistant breast cancer.
Sex, Age, Specimen part, Disease, Cell line, Treatment, Race, Time
View SamplesIdentification of genes up-regulated in ALK-positive and EGFR/KRAS/ALK-negative lung adenocarcinomas.
Identification of genes upregulated in ALK-positive and EGFR/KRAS/ALK-negative lung adenocarcinomas.
Sex, Age, Specimen part
View SamplesGene expression was examined in granulosa cells and oocytes in various stage of follicle and in vitro grown oocytes and granulosa cells complexes in sus scrofa.
Gene expression patterns in granulosa cells and oocytes at various stages of follicle development as well as in in vitro grown oocyte-and-granulosa cell complexes.
Specimen part
View SamplesOne hundred and seven lung Squamous Cell Carcinomas collected from early stage (stage I+II; AJCC 7th edition) patients at the National Cancer Center Hospital (Tokyo, Japan) between 1997 and 2008 were hybridized to the Human Transcriptome (HT) Array 2.0
A Two-Gene Prognostic Classifier for Early-Stage Lung Squamous Cell Carcinoma in Multiple Large-Scale and Geographically Diverse Cohorts.
Sex, Age, Specimen part
View SamplesGene expression analysis to compare control cells and sorted cells
Identification of two major autoantigens negatively regulating endothelial activation in Takayasu arteritis.
Specimen part
View SamplesThis SuperSeries is composed of the SubSeries listed below.
Recurrent mutations in multiple components of the cohesin complex in myeloid neoplasms.
Specimen part, Disease, Cell line
View Samples