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Gene expression analysis of airway epithelial cells exposed to flagellin via RNA-seq and microarray

ABSTRACT: Gene expression analysis of airway epithelial cells exposed to flagellin via RNA-seq and microarray

PROVIDER: PRJNA239683 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR1179090.fastq.gz	Fastqsanger.gz
	SRR1179100.fastq.gz	Fastqsanger.gz
	SRR1179110.fastq.gz	Fastqsanger.gz
	SRR1179120.fastq.gz	Fastqsanger.gz
	SRR1179130.fastq.gz	Fastqsanger.gz

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Gene expression analysis of airway epithelial cells exposed to flagellin via microarray

Project description:Airway epithelial cells (AEC) are critical components of the inflammatory and immune response during exposure to pathogens. AECs in monolayer culture and differentiated epithelial cells in air-liquid interface (ALI) represent two distinct and commonly used in vitro models, yet differences in their response to pathogens have not been investigated. In this study, we compared the transcriptional effects of flagellin on AECs in monolayer culture versus ALI culture using exon microarrays and RNAsequencing. We found that AECs cultured in monolayer and ALI have strikingly different transcriptional states at baseline. When challenged with flagellin, monolayer AEC cultures greatly increased transcription of numerous genes mapping to wounding response, immunity and inflammatory response. In contrast, AECs in ALI culture had an unexpectedly muted response to flagellin, both in number of genes expressed and relative enrichment of inflammatory and immune pathways. In conclusion, In vitro culturing methods have a dramatic effect on the transcriptional profile of AECs at baseline and after stimulation with flagellin. These differences suggest that epithelial responses to pathogen challenges are distinctly different in culture models of intact and injured epithelium.

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Gene expression analysis of airway epithelial cells exposed to flagellin via RNA-seq

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Transcriptomic profile of Calu-3 airway epithelial cells in response to flagellin

Project description:The purpose of the study is to compare the transcriptomic profile of the airway epithelial cell line Calu-3 in reponse to Pseudomonas aeruginosa virulence factor flagellin. CF is caused by mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene. CFTR gene in Calu-3 cells was knockdown by CRISPR-Cas9 (TS) and a control cell line (CTL) was aslo generated using a non-targeting guide RNA. CTL and CFTR KD Calu3-cells were grown on Transwell filters, polarized at the air-liquid interface and exposed to flagellin for 6 hours. We considered 2 non-stimulated (ND) conditions: CTL-NS and TS-NS, and 2 conditions stimulated with flagellin (Flag): CTL-Flag and TS-Flag.

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Project description:Here, we aim to understand the role of P38 in the airway epithelial cells response to flagellin. We report the gene expression profile of pig airway epithelial cells cultured under air-liquid conditions, pre-incubated or not with the P38 inhibitor SB203580 (20 µM, Tocris Bioscience) for 1 h and then stimulated with 100 ng/ml of a mutated flagellin (FliCΔ174-400) for 2h or 24h. Our data show that flagellin stimulation induce the expression of pro-inflammatory cytokines mainly through the NFkB pathway, with little impact of P38 inhibition in the cells response.

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Bronchial epithelial cells transcriptomics upon flagellin stimulation (Microarray)

Project description:Innate immune memory is a new concept describing ability of innate myeloid and lymphoid cells to gain memory characteristics after transient stimulation, resulting in an nonspecific modified response upon secondary challenge. In recent work, we demonstrated that bronchial epithelial cells, non immune cells, that are the first immune cells of the lower respiratory tract to encounter pathogens were capable of innate immune memory after pre-exposure to Pseudomonas aeruginosa flagellin. In the present study we identified chromatin modifications induced by flagellin pre-exposure that sustain reprogramming of transcriptional patterns. We researched chromatin structure modifications responsible for the modulation of gene expression and thus of innate memory induced by the exposure of cells to flagellin.

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