Title: Diesel exhaust particles dysregulate multiple immunological pathways in murine macrophages: Lessons from microarray and scRNA-seq technologies.

Authors: Bhetraratana, May; Orozco, Luz D; Hong, Jason; Diamante, Graciel; Majid, Sana; Bennett, Brian J; Ahn, In Sook; Yang, Xia; Lusis, Aldons J; Araujo, Jesus A

Published In Arch Biochem Biophys, (2019 12 15)

Abstract: Exposure to ambient particulate matter has been shown to promote a variety of disorders, including cardiovascular diseases predominantly of ischemic etiology. However, the mechanisms linking inhaled particulates with systemic vascular effects, resulting in worsened atherosclerosis, are not well defined. We assessed the potential role of macrophages in translating these effects by analyzing gene expression patterns in response to diesel exhaust particles (DEP) at the average cell level, using Affymetrix microarrays in peritoneal macrophages in culture (in vitro), and at the individual cell level, using single-cell RNA sequencing (scRNA-seq) in alveolar macrophages collected from exposed mice (in vivo). Peritoneal macrophages were harvested from C57BL/6J mice and treated with 25 μg/mL of a DEP methanol extract (DEPe). These cells exhibited significant (FDR < 0.05) differential expression of a large number of genes and enrichment in pathways, especially engaged in immune responses and antioxidant defense. DEPe led to marked upregulation of heme oxygenase 1 (Hmox1), the most significantly upregulated gene (FDR = 1.75E-06), and several other antioxidant genes. For the in vivo work, C57BL/6J mice were subjected to oropharyngeal aspiration of 200 μg of whole DEP. The gene expression profiles of the alveolar macrophages harvested from these mice were analyzed at the single-cell level using scRNA-seq, which showed significant dysregulation of a broad number of genes enriched in immune system pathways as well, but with a large heterogeneity in how individual alveolar macrophages responded to DEP exposures. Altogether, DEP pollutants dysregulated immunological pathways in macrophages that may mediate the development of pulmonary and systemic vascular effects.

PubMed ID: 31568751

MeSH Terms: Air Pollutants/toxicity*; Animals; Antioxidants/metabolism; Immunity, Innate/drug effects; Immunity, Innate/genetics; Macrophages, Alveolar/drug effects; Macrophages, Alveolar/immunology; Macrophages, Alveolar/metabolism; Macrophages, Peritoneal/drug effects; Macrophages, Peritoneal/immunology; Macrophages, Peritoneal/metabolism; Macrophages/drug effects*; Macrophages/immunology*; Macrophages/metabolism; Male; Mice; Mice, Inbred C57BL; Oligonucleotide Array Sequence Analysis*; RNA, Small Cytoplasmic/genetics*; RNA-Seq*; Vehicle Emissions/toxicity*