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Bronchoalveolar lavage of healthy human adults

ABSTRACT: Bronchoalveolar lavage of healthy human adults

PROVIDER: PRJNA637757 | ENA |

REPOSITORIES: ENA

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Bronchoalveolar lavage of healthy human adults after LPS exposure

Project description:Bronchoalveolar lavage of healthy human adults after LPS exposure

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Bronchoalveolar lavage of healthy human adults

Project description:Rationale: Airspace macrophages are the most abundant cell in airspaces and are viewed as a homogeneous population during health. Single cell RNA sequencing allows for examination of transcriptional heterogeneity between cells and between individuals. Understanding the conserved repertoire of airspace leukocytes during health is essential to understanding cellular programing during disease. Objective: We sought to determine the transcriptional heterogeneity of human bronchoalveolar lavage cells in healthy adults. Methods: Ten healthy subjects underwent bronchoscopy. Cells obtained from lavage fluid were subjected to single cell RNA sequencing. Unique cell populations and putative functions were identified. Transcriptional profiles were compared across individuals. Measurements and Main Results: Based on transcriptional profiling we identify highly conserved macrophage, monocyte-like, lymphocyte, dendritic cell, and cycling cell populations. We define two unique subgroups of resident airspace macrophages - one defined by a pro-inflammatory profile and one by metallothionein gene expression. We identify distinct subsets of monocyte-like cells and directly compared them to peripheral blood mononuclear cells. Finally, we compare global macrophage and monocyte programing between male and female subjects. Conclusions: Healthy human airspaces contain multiple populations of leukocytes that are highly conserved between individuals and between the sexes. Resident macrophages comprise the largest population and include novel subsets defined by inflammatory and metal-binding gene signatures. Monocyte-like cells within the airspaces are transcriptionally distinct from circulating blood cells and include a rare population defined by expression of cell-matrix interaction genes. This study is the first to define airspace immune cell heterogeneity and identifies three previously unrecognized myeloid cell subsets.

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Bronchoalveolar lavage of healthy human adults after LPS exposure

Project description:Rationale: Macrophages are both drivers of dysregulated inflammation and essential for healthy repair following acute lung injury. Distinct subsets of human macrophages have been associated with lung pathology but whether unique cellular programs are causally linked to disease or merely represent a conserved response to tissue damage is unknown. Objectives: We sought to identify the transcriptional heterogeneity and cellular programing of airspace macrophages (AM) during normal lung repair in a human model of self-resolving acute lung injury. Methods: Fifteen subjects underwent bronchoscopic lavage before and at a pre-assigned time point after endobronchial exposure to bacterial endotoxin. Cells from lavage were subjected to single-cell RNA sequencing and a longitudinal assessment of AM programing during resolution of inflammation and lung repair was performed. Measurements and Main Results: We identify transcriptionally distinct subsets of AM present at all time points, in all subjects. We focus on two populations that increase following inflammation, one which aligns closely with classical circulating monocytes (MoAM) and one with interstitial macrophages (IMAM). Comparison of subset-specific markers to those identified in disease states reveals that IMAM express many so-called “pathogenic” macrophage markers during both health and inflammation. Conclusions: By applying a uniform inflammatory stimulus to healthy adults and examining BAL cells obtained at precise time points following inflammation, we construct a time-resolved kinetic of AM transcriptional programing during inflammation and normal lung repair. Our data suggest that IMAM are similar to cells identified at increased frequency in disease states and may reflect a conserved cellular response to tissue injury.

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Project description:We profiled DNA methylation, mRNA expression, and miRNA expression from bronchoalveolar lavage cells obtained from 64 sarcoidosis subjects and 16 healthy controls.

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