Differential transcriptional responses of murine alveolar macrophages to elongated mineral particles of asbestiform and non-asbestiform varieties
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ABSTRACT:
PROVIDER: PRJNA663339 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR12638361_1.fastq.gz | Fastqsanger.gz | |||
| SRR12638361_2.fastq.gz | Fastqsanger.gz | |||
| SRR12638362_1.fastq.gz | Fastqsanger.gz | |||
| SRR12638362_2.fastq.gz | Fastqsanger.gz | |||
| SRR12638363_1.fastq.gz | Fastqsanger.gz |
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Project description:Occupational exposures to asbestiform elongated mineral particles (EMPs) may lead to diffuse fibrosis, lung cancer, malignant mesothelioma and autoimmune diseases. Cleavage fragments (CF) are chemically identical to asbestiform varieties of parent mineral, but there is no consensus on whether to treat them as asbestos from toxicological and regulatory standpoints. Alveolar macrophages (AM) are the first-responders to inhaled particulates, participating in clearance, and activating the other resident and recruited immunocompetent cells, which has an impact on the long-term outcomes. In the current study we addressed the question of how asbestiform vs. non-asbestiform EMPs affect AM responses. Max Planck Institute (MPI) cells, a non-transformed mouse line that has AM phenotype and genotype, were treated with surface area-equivalent concentrations of respirable asbestiform and non-asbestiform riebeckite/tremolite EMPs for 24h. Detectable changes in macrophage gene expression happen within 6-24 hours post-stimuli (Schneider et al., 2019), thus we performed the next generation RNA sequencing of the MPI cells 24 hours post exposure to surface area-equivalent doses of riebeckite and tremolite EMPs to see if AM immediate transcriptional reprogramming patterns are intrinsic for different minerals and/or crystallization habit.
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Project description:Differential transcriptional responses of murine alveolar macrophages to elongated mineral particles of asbestiform and non-asbestiform varieties
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2013-01-31 | E-GEOD-40397 | biostudies-arrayexpress
Project description:Exposure to particulate matter (PM) is consistently associated with increased morbidity and mortality attributable in part to respiratory illnesses. The alveolar macrophage (AM) is one of the cell types in the lung constantly exposed to and activated by ambient pollutants. Upon contact with environmental particulate pollutants, AM produces reactive oxygen species (ROS) and antioxidant enzymes, but the scope of this oxidative stress response induced by PM remains poorly defined. In this study, we used microarray analysis to determine the gene expression profile in human alveolar macrophages upon exposure to PM and sought to gain more insight into the global response of pro- and anti-oxidant enzymes to PM exposure. Human AM were obtained by bronchoscopy from normal individuals. They were then incubated with Chapel Hill PM2.5 (1 ïg/ml) or vehicle for 4 hours (n = 6 independent samples each). mRNAs were extracted, amplified and hybridized to Agilent Human 1A microarray. Differentially expressed genes were identified by Statistical Analysis for Microarrays (SAM) with a FDR of 10% and a P ⤠0.05. Significant genes were also mapped with Gene Ontology (GO) based on their molecular function. We found 34 and 41 up- and down-regulated genes respectively. Of these, 22 genes (~30%) were involved in metal binding and 14 genes were linked to oxidative stress, including 5 metallothionein-1 (MT1) isoform genes. In lung cells, addition of MT-1F in the medium attenuated PM2.5-induced H2O2 production while knockdown of MT1F gene expression increased H2O2 and IL-6 release induced by PM2.5. Our microarray experiments provided a global view of gene expression after in vitro PM2.5 exposure in human AM. The expression profile was most notable for differential expression of genes related to metal binding and oxidative stress, especially upregulation of MT1 isoform genes. Our findings suggest that metals associated with PM, e.g., zinc, copper, and arsenic, induce MT-1 and may be the primary mediators for PM-induced oxidative stress. Keywords: in vitro exposure, PM2.5 particle treatment, human alveolar macrophages Total RNA was isolated from alveolar macrophages from 6 subjects were treated in vitro with PM2.5 particles or vehicle control for 4 hrs. All particle-treated samples were co-hybridized with the common reference sample (pool of all 6 vehicle-treated samples) twice with Cy3 and Cy5 dyes flipped (12 hybridizations). Three samples were also co-hybridized with its own individual control (vehicle-treatment for the same subject) with Cy3 and Cy5 dyes flipped (6 additional hybridizations).
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