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Spatial analysis of young and old murine lungs exposed to ozone and fibrogenic stress [spatial_ozone]

ABSTRACT: Ozone (O3) is a ubiquitous pollutant known to produce acute and transient inflammation through oxidative injury and inflammation. These effects are exacerbated in susceptible populations, such as the elderly and those exhibiting genetic mutations in central nodes of pulmonary function. To comprehend the impact of these predisposing factors, the present study examines structural, mechanical, and immunological responses to single acute O3 exposure (0.8 ppm, 3h) in young (12 week old) and old (80 week old) mice. We therefore examined the impact of ozone in a surfactant competent and mutant murine line. The latter models a clinically relevant mutation in the gene encoding for the alveolar epithelial type 2 specific surfactant protein C. Ozone exposure produced focal injury localized at the terminal bronchiole-to-alveolar junctions and enlarged alveoli in aged SP-C mutant lungs. This histological feature was used to select regions of interest for spatial transcriptomics. Paraffin sections of 4 mice were prepared: a young/12wk SP-C wild type air control, a young/12wk SP-C wild type ozone (0.8 ppm, 3h), a young/12wk SP-C mutant ozone, and an old/80wk ozone. Immunofluorescence for DNA, CD45, CD68 was performed to identify macrophages (CD45+CD68+) and non-immune ("other", DNA+CD45-) cells within healthy (air control only), peri-injured and injured regions of the lung. A total of 24 regions (680um x 700um in size) were selected from CD68+ macrophages and CD45- non immune/"other" cells. RNA sequencing was then performed. Principal component and pathway analysis revealed spatial heterogeneity in response to ozone, as well as a senescence signature in the aging lungs. These results illustrate regional and age dependent heterogeneity in immune and non-immune cells of the lung during an acute exposure to air pollution.

ORGANISM(S): Mus musculus

PROVIDER: GSE273048 | GEO | 2025/01/10

REPOSITORIES: GEO

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Project description:In this study, integrated transcriptomics, proteomics and metabolomics approaches were applied to investigate the molecular responses of O3 in the leaves of two-weeks old rice (cv. Nipponbare) seedlings exposed to 0.2 ppm O3 for a period of 24 h. Based on the morphological alteration of O3-exposed rice leaves, transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that O3 triggers a chain reaction of altered gene, protein and metabolite expressions involved in multiple cellular processes in rice. Also investigated were the molecular responses in the leaves of two-weeks old rice (cv. Nipponbare) seedlings under continuous light and pure air (as a positive control for ozone exposure experiments) for a period of 24 h. Transcript profiling of rice genes was performed in leaves exposed for 1, 12 and 24 h using rice DNA microarray chip, proteomics and metabolomics. This systematic survey showed that continuous light and growth for 24 h also triggers a chain reaction of altered gene expressions involved in multiple cellular processes in rice, but different from those against ozone, in general. Experiment Overall Design: Ozone-treated Samples: An in vivo 14-days-old rice seedling model system was used, where whole seedlings were used for exposure/fumigation to the ozone (0.2 ppm). Dye-swap or reverse labeling with Cy3 and Cy5 dyes procedure was applied followed by hybridization and wash processes, and the hybridized microarrays (G4138A) were scanned using a GenePix microarray scanner followed by the Gene Pix 4000 analysis application program for image analysis and data extraction processes. The GeneSpring Ver. 4 software was used for normalization. Experiment Overall Design: Control Samples: An in vivo 14-days-old rice seedling model system was used, where whole seedlings were used for exposure to continuous light and pure air. Dye-swap or reverse labeling with Cy3 and Cy5 dyes procedure was applied followed by hybridization and wash processes, and the hybridized microarrays (G4138A) were scanned using a Standard protocol of Agilent DNA Microarray Scanner and data processing was done by Feature Extraction software Version 8.1 from Agilent Technologies.

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Spatial analysis of young and old murine lungs exposed to ozone and fibrogenic stress [spatial_ozone]

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