Expansion of SFTPC+ cells from human induced pluripotent stem cell-derived CPMhigh progenitor cells by forming alveolar organoids
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE90591 | GEO | 2017/10/02
SECONDARY ACCESSION(S): PRJNA355125
REPOSITORIES: GEO
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Project description:Microarray analysis was performed on P0 SFTPC+ cells, P0 SFTPC- cells and CPMhigh progenitor cells derived form human induced pluripotent stem cells (B2-3 SFTPC-GFP reporter hiPSCs), respectively.
2017-10-02 | GSE90590 | GEO
Project description:Mutations in the gene encoding surfactant protein C (SFTPC) are associated with interstitial lung disease in children and adults. To assess the natural history of disease, we knocked-in a familial, disease-associated SFTPC mutation, L188Q (L184Q in mice), into the mouse Sftpc locus. Expression of the L184Q allele (LQ) resulted in formation of an unstable, misfolded proprotein (proSP-CLQ) that was rapidly degraded by the proteasome pathway. Translation of proSP-CLQ exceeded that of proSP-CWT in neonatal AT2 cells and was associated with transient activation of oxidative stress and apoptosis leading to impaired expansion of AT2 cells during postnatal alveolarization. Consequently, adult mutant mice demonstrated a decrease in the number of AT2 cells with no effect on lung homeostasis; however, lung regeneration in response to bleomycin challenge was substantially reduced in mutant mice. Collectively, these data support the hypothesis that susceptibility to disease in adult mutant mice is established during postnatal lung development and is associated with a permanent reduction in AT2 cell numbers.
2020-08-19 | MSV000085976 | MassIVE
Project description:Dyskeratosis congenita (DC) is a rare genetic disorder characterized by deficiencies in telomere maintenance leading to very short telomeres and the premature onset of certain age_Related diseases, including pulmonary fibrosis (PF). PF is thought to derive from epithelial failure, particularly that of type II alveolar epithelial (AT2) cells, which are highly dependent on Wnt signaling during development and adult regeneration. We use human iPSC-derived AT2 (iAT2) cells to model how short telomeres affect AT2 cells. Cultured DC mutant iAT2 cells accumulate shortened, uncapped telomeres and manifest defects in the growth of alveolospheres, hallmarks of senescence, and apparent defects in Wnt signaling. The GSK3 inhibitor, CHIR99021, which mimics the output of canonical Wnt signaling, enhances telomerase activity and rescues the defects. These findings support further investigation of Wnt agonists as potential therapies for DC related pathologies.
2021-05-07 | GSE160871 | GEO
Project description:The pulmonary alveolar epithelium mainly composed of two types of epithelial cells: alveolar type I (AT1) and type II (AT2) cells. AT2 cells are the alveolar stem cells, and can differentiate into AT1 cells post-pneumonectomy (PNX). Here, we found that, compared with control mice (Sftpc-CreER; Cdc42flox/+; Rosa26-mTmG) at post-PNX day 21, Cdc42 AT2 null mice (Sftpc-CreER; Cdc42flox/-; Rosa26-mTmG) at post-PNX day 21 undergone fibrotic change. By using 10X genomics “Chromium Single Cell” technology, we performed single-cell RNA-seq analyses of AT2 cells of sham treated control mice (C0), AT2 cells of control mice at post PNX day 21 (C21) , AT2 cells of sham treated Cdc42 AT2 null mice (N0), and AT2 cells of Cdc42 AT2 null mice at post PNX day 21 (N21). The study identified a specific gene signature in AT2 cells of Cdc42 AT2 null mice at post PNX day 21 which is related to the fibrosis phenotype of Cdc42 AT2 null mice.
2020-01-03 | GSE138585 | GEO
Project description:Mutations in the gene encoding surfactant protein C (SFTPC) are associated with interstitial lung disease in children and adults. To assess the natural history of disease, we knocked-in a familial disease-associated SFTPC mutation, L188Q (L184Q in mice), into the mouse Sftpc locus. Expression of L184Q (LQ) resulted in formation of an unstable, misfolded proprotein (proSP-CLQ) that was rapidly degraded by the proteasome pathway. Importantly, expression of misfolded proSP-CLQ was associated with a decrease in AT2 cells in adult mutant mice, with no effect on lung homeostasis; however, lung regeneration in response to bleomycin challenge was substantially reduced in mutant mice. Translation of proSP-CLQ exceeded that of proSP-CWT in neonatal AT2 cells and was associated with activation of oxidative stress and apoptosis leading to impaired expansion of AT2 cells during postnatal alveolarization. Collectively, these data support the hypothesis that susceptibility to disease in adult mutant mice is established during postnatal lung development and is associated with a reduction in AT2 cell numbers.
2021-06-30 | GSE155814 | GEO
Project description:During human lung development the distal tip epithelial cells act as multipotent progenitors. From ~15 pcw (post conception weeks) the human tip epithelium retains progenitor marker expression, but also upregulates markers of the AT2 lineage. Immature AT2 cells appear in the tissue from 17 pcw. We have recently shown that 16-22 pcw epithelial tip cells can be expanded as organoids and differentiated to AT2 cells. However, the differentiated AT2 cells were not proliferative, limiting their use for functional studies and genetic manipulation. We have now developed a highly robust, efficient, and scalable culture condition (AT2 medium) that induces the differentiation of 16-22 pcw fetal lung tip cells into mature AT2 cells which grow as expandable 3D organoids. In this study, we characterise the fetal-derived AT2 cells, showing that they are stable over long-term passaging, efficiently process and secrete surfactant, and can differentiate into AT1 cells in vitro and in mouse lung transplantation assays. Forward genetic screen identifies candidate effectors of SFTPC trafficking which we validate using CRISPR interference (CRISPRi) in the AT2 organoids. We show that the trafficking of SFTPC requires ubiquitination by HECT domain E3 ligases, particularly Itch, and that their depletion phenocopies the pathological redistribution seen for the SFTPCI73T variant.
2023-08-31 | GSE237359 | GEO
Project description:Rationale: Aryl hydrocarbon receptor (AhR), a ligand-activated transcription factor, has been considered as an important regulator for immune diseases. We have previously shown that AhR protects against allergic airway inflammation. The underlying mechanism, however, remains undetermined. Objectives: We sought to determine whether AhR specifically in Type II alveolar epithelial cells (AT2) modulates allergic airway inflammation and its underlying mechanisms. Methods: The role of AhR in AT2 cells in airway inflammation was investigated in a mouse model of asthma with AhR conditional knock out mice in AT2 cells (Sftpc-Cre;AhRflox/flox). The effect of AhR on allergen-induced autophagy was examined by both in vivo and in vitro analyses. The involvement of autophagy in airway inflammation was analyzed by using autophagy inhibitor chloroquine. The AhR-regulated gene profiling in AT2 cells was also investigated by RNA-seq analysis. Results: Sftpc-Cre; AhRflox/flox mice showed exacerbation of allergen-induced airway hyperresponsiveness and airway inflammation with elevated Th2 and airway epithelial-derived cytokines in bronchoalveolar lavage fluid (BALF). Notably, an increased allergen-induced autophagy was observed in the lung tissues of Sftpc-Cre; AhRflox/flox mice when compared with wild-type mice. Further analyses suggested a functional axis of AhR-TGF-β1 that is critical in driving allergic airway inflammation through regulating allergen-induced cellular autophagy. Furthermore, inhibition of autophagy suppressed allergic airway inflammation with decreased Th2 and epithelial cell-derived cytokines in BALFs. Additionally, RNA-seq analysis suggests that autophagy is one of the major pathways and CALCOCO2/NDP52 and S1009 are major autophagy-associated genes in AT2 cells that contribute to the AhR-mediated allergic airway inflammation. Conclusion: These results suggest that AhR in AT2 cells functions as a protective mechanism against allergic airway inflammation through controlling cell autophagy.
2022-06-13 | GSE205818 | GEO
Project description:We stepwisely induced SFTPC+ cells from hiPSCs via CPM-high progenitor cells with or without coculturing human fetal lung fibroblasts. Single-cell RNA-seq of CPM-high progenitor and hiPSC-derived SFTPC+ cells demonstrated their differentiation process and cellular heterogeneity.
2017-10-02 | GSE90813 | GEO
Project description:Rationale: Alveolar epithelial type 2 (AT2) cells are stem/progenitor cells in the adult lung and their maintenance and regulation is achieved through their interaction with mesenchymal cells which constitute their niches. However, the precise identity of the niche cells is still elusive. Objectives: To characterize the niche cells capable of supporting the self-renewal of AT2 stem cells in the murine model. Methods: The alveolar organoid model was used to test the efficiency of different subpopulations of mesenchymal cells isolated by FACS from well-established (PdgfraGFP, Axin2LacZ, Fgf10LacZ) reporter mice to support the self-renewal of mature AT2 cells. The reporter-positive cells where pre-sorted based on Cd45neg Cd31neg Epcamneg Sca1pos. Additional selection was achieved using LipidTOX staining for cells containing high level of neutral lipids. Organoid size and colony formation efficiency after 2 weeks in culture were quantified. RNAscope combined with immunofluorescence on adult lungs. qPCR, gene array were used to characterize the niche cells. Measurements and main results: A subset of Sca1+; Fgf10-expressing cells positive for LipidTox staining in the distal mesenchyme is efficient in the self-renewal and differentiation of AT2 stem cells. Co-staining of adult lung by using ISH and IF staining for Fgf10 and Sftpc respectively, indicate that 28% of Fgf10+ cells are located to close to AT2 cells. These cells do not overlap with Fgf7-expressing cells. Gene array comparing MANC (Cd45Neg Cd31Neg EpcamNeg Sca1Pos Axin2LacZ/FDGPos) with Fgf10Pos Lipofibroblasts (Cd45Neg Cd31Neg EpcamNeg Sca1Pos Fgf10LacZ/FDGPos) support that these two cell subsets are different and express differential markers that can be further used for the respective characterization of these cells. Conclusions: We have demonstrated for the first time that Fgf10Pos LIF cells play important role to support AT2 stem cells in terms of self-renewal and differentiation toward the AT1 cell type. These cells appear different from the previously characterized MANC and display a similar activity in eliciting AT2 self-renewal.
2021-12-01 | GSE162859 | GEO
Project description:Alveolar epithelial type 2 (AT2) cells are facultative progenitor cells that drive adult alveolar regeneration after acute lung injury. Using transcriptomic analyses from in vivo mouse injury models, we define the role of Tfcp2l1 in regulating AT2 cell behavior during lung regeneration.
2023-09-12 | GSE204786 | GEO