Temporal retinal transcriptome analysis in Staphylococcus aureus endophthalmitis
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ABSTRACT:
PROVIDER: PRJNA824511 | ENA |
REPOSITORIES: ENA
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Project description:Bacterial endophthalmitis remains a devastating inflammatory disease associated with permanent vision loss. The mechanisms regulating inflammation to protect the eye remain elusive. Therefore, assessing the host immune response in bacterial endophthalmitis may provide new therapeutic targets. We used microarrays to determine the global profiling of up-regulated and downregulated gene expression in bacterial endophthalmitis.
2022-04-08 | GSE200443 | GEO
Project description:Purpose: Endophthalmitis is a serious complication of intraocular surgery due to the risk of irreversible retinal damage. The heterogeneity of retinal cells necessitates analysis at the single-cell level to uncover the detailed mechanisms of infection response. Using a mouse model of bacterial endophthalmitis, we investigated the affected pathways of resident and infiltrating cell types in the retina. Methods: A methicillin-sensitive strain of Staphylococcus aureus was isolated from a patient with endophthalmitis. Adult C57Bl/6J mice received an intravitreal injection of phosphate-buffer solution (PBS) with or without 5000 CFU S. aureus (n=3 per group). 24 hours later, retinas were isolated and single-cell suspensions were sent to the Penn Genomic Core for sequencing with an Illumina NovaSeq 6000. After standard pre-processing of the data, differential genes and pathways were identified for each cell type (adjusted p < 0.01, log2FC > 1 or < -1). Results: Our analysis identified all expected retinal cell types, including a population of infiltrating neutrophils in the infected group. We surveyed genes known to be upregulated at the bulk-retina level in this model (e.g. Tlr2, Nlrp3, Il1b), and found that infiltrating cells mainly drove this expression. A few genes changed across almost all cell types, including upregulation of Hsph1 and Stat3. Muller glia downregulated Gpx4 while upregulating Acsl4 and iron importers Tfrc, Zip14, and Dmt1. Top pathways for macrophages/microglia included chemotaxis, cell-cell adhesion, and wound healing. Vascular cells upregulated angiogenesis-related genes. Cellular respiration was a commonly affected pathway across several neuronal populations, with most genes decreasing. Conclusions: This study sought to advance our understanding of the pathobiology of bacterial endophthalmitis. Muller glia appear to be undergoing ferroptosis, potentially while activating a program to sequester iron away from bacteria. Decreased cellular respiration may indicate hypoxia among neurons. Our results reveal several trends in the retinal response to infection, including iron dysregulation and hypoxia. Understanding these cell-type-specific responses to endophthalmitis may help design therapies to combine with antibiotics.
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Project description:Staphylococcus aureus (S. aureus) is an important human and animal pathogen, multiply resistant strains are increasingly widespread, new agents are needed for the treatment of S. aureus. sodium houttuyfonate has potent antimicrobial activity against S. aureus. We employed Affymetrix Staphylococcus aureus GeneChipsTM arrays to investigate the global transcriptional profiling of Staphylococcus aureus ATCC25923 treated with sodium houttuyfonate. Keywords: gene expression array-based, count
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Project description:Staphylococcus aureus (S. aureus) is an important human and animal pathogen, multiply resistant strains are increasingly widespread, new agents are needed for the treatment of S. aureus. Berberine chloride (BBR), a natural plant product, has potent antimicrobial activity against S. aureus. We employed Affymetrix Staphylococcus aureus GeneChipsTM arrays to investigate the global transcriptional profiling of Staphylococcus aureus ATCC25923 treated with berberine chloride. Keywords: gene expression array-based, count
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