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Hedgehog gradient in the stroma maintains niche diversity and organ function

ABSTRACT: Hedgehog gradient in the stroma maintains niche diversity and organ function

PROVIDER: PRJNA398101 | ENA |

REPOSITORIES: ENA

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Hedgehog gradient in the stroma maintains niche diversity and organ function [scRNA-seq]

Project description:Hedgehog gradient in the stroma maintains niche diversity and organ function [scRNA-seq]

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Hedgehog gradient in the stroma maintains niche diversity and organ function [RNA-seq]

Project description:We disrupted the spatial Hh activation gradient in murine lung by inducing the constitutively active form of the Hh effector, Smo (SmoM2), throughout the entire GLI2+ stroma and performed RNA-seq on the bulk GLI2+ population. Whole adult murine lung tissue was dissociated to single cells and subjected to fluorescence activated cell sorting (FACS) to select all live GLi2+ cells. The bulk RNA-sequencing library was then subsequently generated. Cells were sequenced at a depth of average of 45 million reads/sample. The results reveal that expansion of Hh activation in the GLI2+ domain in the murine lung resulted in upregulation of genes that are expressed in the proximal stroma fibroblasts. Conversely, Hh activation downregulated genes enriched in the distal stroma as well as mesothelial cells.

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Hedgehog gradient in the stroma maintains niche diversity and organ function [scRNA-seq]

Project description:Two single-cell transcriptional analyses were performed on adult human lung distal and proximal stromal cells, respectively. We isolated lung specimen from both the proximal portion enriched in airways and distal portion enriched in alveoli separately. The lung fragments were then dissociated to single cells and FACSorted for stromal cells based on negative expression of epithelial, hematopoietic, and endothelial markers (EPCAM-/CD45-/CD11b-/CD31-). Library preparation was performed separately for the proximal and distal-derived stromal cells so that each subset has distinctive barcodes to determine anatomical origin, and approximately 10,000 cells were captured from each specimen. The results demonstrate that the proximal and distal human lung stromal subsets demonstrate significant overlap of homologous genes with their murine counterparts generated from Gli2+ murine stroma single-cell transcriptional analysis.

2018-08-08 | GSE108288 | GEO

Hedgehog gradient in the stroma maintains niche diversity and organ function

Project description:This SuperSeries is composed of the SubSeries listed below.

2018-08-08 | GSE102592 | GEO

Hedgehog gradient in the stroma maintains niche diversity and organ function [Drop-seq]

Project description:A single-cell transcriptional analysis was performed on GLI2+ stromal cells from the adult murine lung. Whole adult murine lung tissue was dissociated to single cells and subjected to fluorescence activated cell sorting (FACS) to select all live GLI2+ cells. The single cell RNA-sequencing library was then subsequently generated. Cells were sequenced at a depth of ~60,000 reads/cell. We captured approximately 4,600 cells with a median of 2,246 genes detected per cell utilizing a droplet-based barcoding approach to capture single cells for RNA sequencing. The results demonstrate that the GLI2+ population is segregated into distinct stromal subtypes along the proximal-distal axis of the lung with distinct functional contribution to matrix production and paracrine signaling within their distinctive niche.

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The function of the Hedgehog signaling pathway in the colon

Project description:The morphogen Indian Hedgehog plays a very important role during intestinal embryogenesis, but also maintains homeostasis in the adult gut. Intestinal Hedgehog is expressed by the intestinal epithelium and signals in paracrine manner to fibroblasts in the stromal compartment. We studied the colonic changes upon activation of the Hedgehog pathway by deleting the Hedgehog receptor Patched1 in order to alleviate its repressive function.

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Project description:Postnatal tissue quiescence is generally thought to be a default state in the absence of a proliferative stimulus such as injury. We now demonstrate that in the lung, quiescence in the adult is an actively maintained state and is regulated by paracrine hedgehog signaling. Epithelial-specific deletion of Sonic Hedgehog during normal homeostasis results in a proliferative expansion of the adjacent lung mesenchyme. Injury to the lung epithelium results in decreased hedgehog activation, accompanied by proliferative expansion of the adjacent mesenchyme. Moreover, reconstitution of Hedgehog signaling during epithelial injury attenuated the proliferative expansion of the adjacent mesenchyme. Hedgehog signaling maintains lung quiescence by attenuating PDGF signaling through blocking post-translational processing of PDGF receptor Î±/Î² into the mature isoforms. These results indicate that in postnatal tissues, epithelial cells can actively maintain mesenchymal quiescence via paracrine hedgehog activation, and that imbalances in this pathway could lead to aberrant mesenchymal expansion and postnatal disease. Fibroblasts were isolated from mouse lungs and grown in culture in triplicate wells. Samples were treated with vehicle or purmorphamine 5um for 24 hours and RNA was isolated for microarray.

2016-07-03 | E-GEOD-61026 | biostudies-arrayexpress

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