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Age-related changes in the cellular composition and epithelial organization of the mouse trachea

ABSTRACT: We report here senescent changes in the structure and organization of the mucociliary pseudostratified epithelium of the mouse trachea and the main stem bronchi. We confirm previous reports of the graduate appearance of age-related, gland-like structures (ARGLS) in the submucosa, espeically in the intercartilage regions and carina. Immunohistochemistry shows these structures contain ciliated and secretory cells and Krt5+ basal cells, but not the myoepithelial cells or ciliated ducts typical of normal submucosal glands. Data suggests they arise de novo by budding from teh surface epithelium rather than by delayted growth of small or cryptic submucosal glands. In old mice the surface epithelium contains fewer cells per unit length than in young mice and the proportion of Krt5+, p63+ basal cells is reduced in both males and females. However, there appears to be no significant difference in the ability of basal stem cells isolated from individual young and old mice to form clonal tracheospheres in culture or in the ability of the pithelium to repair after damage by inhaled sulfur dioxide. Gene expression analysis by Affymetrix microarray and quantitative PCR, as well as immunohistochemistry and flow sorting studies, are consistent with low-grade chronic inflammation in the tracheas of old versus young mice. The significance of these changes for ARGL formation are not clear since several treatments that induce acute inflammation in young mice did not result in budding of the surface epithelium.

ORGANISM(S): Mus musculus

PROVIDER: GSE55162 | GEO | 2014/02/20

SECONDARY ACCESSION(S): PRJNA238657

REPOSITORIES: GEO

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Project description:We report here senescent changes in the structure and organization of the mucociliary pseudostratified epithelium of the mouse trachea and the main stem bronchi. We confirm previous reports of the graduate appearance of age-related, gland-like structures (ARGLS) in the submucosa, espeically in the intercartilage regions and carina. Immunohistochemistry shows these structures contain ciliated and secretory cells and Krt5+ basal cells, but not the myoepithelial cells or ciliated ducts typical of normal submucosal glands. Data suggests they arise de novo by budding from teh surface epithelium rather than by delayted growth of small or cryptic submucosal glands. In old mice the surface epithelium contains fewer cells per unit length than in young mice and the proportion of Krt5+, p63+ basal cells is reduced in both males and females. However, there appears to be no significant difference in the ability of basal stem cells isolated from individual young and old mice to form clonal tracheospheres in culture or in the ability of the pithelium to repair after damage by inhaled sulfur dioxide. Gene expression analysis by Affymetrix microarray and quantitative PCR, as well as immunohistochemistry and flow sorting studies, are consistent with low-grade chronic inflammation in the tracheas of old versus young mice. The significance of these changes for ARGL formation are not clear since several treatments that induce acute inflammation in young mice did not result in budding of the surface epithelium. Total RNA from distal tracheas and carinas of four young (2 month) and four older (14 month) C57Bl/6 female mice was extracted using QIAshredder and RNeasy Micro Kits (QIAGEN). The quality was checked with a 2100 Bioanalyzer (Agilent Technologies). Total RNA was processed using Ambion MessageAmpTM Premier by the Duke Microarray Facility. Standard Affymetrix protocols and Affymetrix GeneChip® Mouse Genome 430 2.0 Array chips were used to generate .cel files.

2014-02-20 | E-GEOD-55162 | biostudies-arrayexpress

Gene expression data from two murine tracheal cell populations

Project description:The airway epithelium is in contact with the environment and therefore constantly at risk for injury. Basal cells have been found to repair the surface epithelium, but the contribution of other stem cell populations to airway epithelial repair have not been identified. We demonstrated that airway submucosal gland duct cells, in addition to basal cells, survived severe hypoxic-ischemic injury. We developed a method to isolate duct cells from the airway. In vitro and in vivo models were used to compare the self-renewal and differentiation potential of duct cells and basal cells. We found that only duct cells were capable of regenerating submucosal gland tubules and ducts, as well as the surface epithelium overlying the submucosal glands. This is of importance to the field of lung regeneration as determining the repairing cell populations could lead to the identification of novel therapeutic targets and cell-based therapies for patients with airway diseases. Murine proximal airway duct and basal cells were isolated from 8-12 week old male and female mice and FACS sorted. Each sample contains cells that were sorted from a different pool of 10-15 C57Bl/6 mice. RNA was extracted, labeled, and hybridized to Affymetrix Mouse Genome 430 2.0 Array (GPL1261)

2012-05-31 | E-GEOD-28651 | biostudies-arrayexpress

Gene expression data from two murine tracheal cell populations

2012-06-01 | GSE28651 | GEO

Single cell RNA Sequencing Identifies G-protein Coupled Receptor 87 as a Basal Cell Marker Expressed in Distal Honeycomb Cysts in Idiopathic Pulmonary Fibrosis

Project description:Idiopathic Pulmonary Fibrosis (IPF) is a devastating and life-threatening lung disease characterized by epithelial reprogramming and increased extracellular matrix deposition leading to loss of lung function. A prominent histopathological structure in the IPF lung are aberrant bronchioles filled with mucus, referred to as honeycomb cysts. These heterogeneous bronchiolized areas feature clusters of simple epithelium with Keratin (KRT)5+ basal-like cells interspersed with pseudostratified epithelium containing differentiated, hyperplastic epithelial cells as well as aberrant ciliated cells. Recent single-cell RNA sequencing studies of the lung epithelium shed further light into cellular subtypes unique to IPF, including basaloid KRT5-/KRT17+ cells present in the distal lung. However, IPF distal bronchiole cell subtypes still remain poorly characterized and their disease contribution remains underinvestigated. Using single-cell RNA sequencing of enriched EpCAM+ lung cells of the distal IPF lung, we identified G-protein coupled receptor (GPR) 87, as a marker of distal bronchioles and KRT5+ IPF basal cells contributing to impaired airway differentiation and honeycombing.

2022-05-01 | GSE190889 | GEO

Transcription profiling of mouse tracheal basal cells

Project description:Epithelial basal cells (BCs) are an important stem cell population of the airways. We purified BCs from a KRT5-GFP transgenic mouse line and used Affymetrix arrays to compare there gene expression to that of non-BC epithelium. Experiment Overall Design: Three populations of cells were obtained by FACS: 1. KRT5-GFP-, lectin- columnar epithelium. 2. KRT5-GFP-, lectin+ BCs. 3. KRT5-GFP+, lectin+ BCs. Three biological replicates of each sample were hybridized to Affymetrix microarray.

2009-07-11 | E-GEOD-15724 | biostudies-arrayexpress

Progenitor identification and SARS-CoV-2 infection in human distal lung organoids

Project description:The distal lung contains terminal bronchioles and alveoli that facilitate gas exchange. Three-dimensional in vitro human distal lung culture systems would strongly facilitate investigation of pathologies including interstitial lung disease, cancer, and SARS-CoV-2-associated COVID-19 pneumonia. We generated long-term feeder-free, chemically-defined culture of distal lung progenitors as organoids derived from single adult human alveolar epithelial type II (AT2) or KRT5+ basal cells. AT2 organoids exhibited AT1 transdifferentiation potential while basal cell organoids developed lumens lined by differentiated club and ciliated cells. Single cell analysis of basal organoid KRT5+ cells revealed a distinct ITGA6+ITGB4+ mitotic population whose proliferation further segregated to a TNFRSF12Ahi subfraction comprising ~10% of KRT5+ basal cells, residing in clusters within terminal bronchioles and exhibiting enriched clonogenic organoid growth activity. Distal lung organoids were created with apical-out polarity to display ACE2 on the exposed external surface, facilitating SARS-CoV-2 infection of AT2 and basal cultures and identifying club cells as a novel target population. This long-term, feeder-free organoid culture of human distal lung, coupled with single cell analysis, identifies unsuspected basal cell functional heterogeneity and establishes a facile in vitro organoid model for human distal lung infections including COVID-19-associated pneumonia.

2020-11-01 | GSE106850 | GEO

Gene expression data from mouse tracheal epithelial cells isolated before and 48hrs after SO2 injury

Project description:The conducting airway epithelium of the rodent and human lung is made up of about equal proportions of ciliated and secretory cells. In addition, in regions where the epithelium is pseudostratfied, ~30% of the epithelium consists of undifferentiated basal cells (BCs). Evidence suggests that these BCs are multipotent stem cells that can self renew over the long term and give rise to both ciliated and secretory lineages. The goal of this project is to identify cellular and molecular mechanisms by which the basal cells normally maintain the epithelium and repair it after injury. We used Affymetrix microarray analysis to compare transcripts in tracheal epithelium before and after SO2 injury. Mice tracheal epithelium and mesenchyme were separate for RNA extraction before and 48hrs after SO2 injury. Cells from tracheas of 4 male C57Bl/6 mice were pooled for each biological experiment. The experiments were repeated three times for triplicate samples.

2016-07-03 | E-GEOD-69056 | biostudies-arrayexpress

microRNAs signatures of Xenopus laevis embryo epidermis at stage 11 (non ciliated) and 26 (ciliated) using high throughput sequencing

Project description:Epidermis of Xenopus embryos forms a mucociliary epithelium constituted of basal, scattered, secreting and ciliated cells and is histologically similar to human airway mucociliary epithelium. We compared microRNAs signatures of epidermis of Xenopus embryos at stage 11.5 (gastrula, non ciliated epidermis) and at stage 26 (tailbud, ciliated epidermis). 2 technical replicates of a pool of 50 explants for each stage 11.5 (non ciliated) and 26 (ciliated) of Xenopus laevis development

2011-06-06 | E-GEOD-22146 | biostudies-arrayexpress

microRNAs signatures of Xenopus laevis embryo epidermis at stage 11 (non ciliated) and 26 (ciliated) using high throughput sequencing

2011-06-07 | GSE22146 | GEO

Single cell-RNA-seq for airway epithelial cells

Project description:Outputs from single-cell RNA-seq sequenced reads from isolated mouse epithelial cells from homeostatic submucosal glands and Naphthalene-injured surface epithelium of adult mouse airway (P70)

2018-03-09 | GSE111598 | GEO

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