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RNA sequencing of blood brain barrier spheroids exposed to either Neisseria meningitidis or Borrelia bavariensis in in vitro conditions

ABSTRACT: RNA sequencing of blood brain barrier spheroids exposed to either Neisseria meningitidis or Borrelia bavariensis in in vitro conditions

PROVIDER: PRJEB67319 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR12102468.fastq.gz	Fastqsanger.gz
	ERR12102469.fastq.gz	Fastqsanger.gz
	ERR12102470.fastq.gz	Fastqsanger.gz
	ERR12102471.fastq.gz	Fastqsanger.gz
	ERR12102472.fastq.gz	Fastqsanger.gz

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RNA sequencing of blood brain barrier spheroids exposed to either Neisseria meningitidis or Borrelia bavariensis in in vitro conditions

Project description:Neisseria meningitidis and Borrelia bavariensis are the causative agents of invasive meningococcal disease and Lyme neuroborrelosis. Before infecting the brain parenchyma the bacterial pathogens should cross the human blood-brain barrier (BBB). To understand the response of BBB against the invading pathogens, in vitro BBB spheroid model was generated by co cultivating human brain microvascular endothelial cells, pericytes and astrocytes in low adhesion conditions. Spheroids (N= 36) were infected with either N. meningitidis (6 x 10e4, MOI 1:4) or B bavariensis(1.5 x 10e5, MOI 1:10) for 3h. Two separate control groups of spheroids (N=36) without any infection were included in the study. After 3h of treatment (infection/no infection), 12 spheroids were pooled to form one biological sample and such 3 biological replicates per treatment were sampled for total RNA extraction. Thereafter, 12 cDNA libraries were prepared using QuantSeq FWD 3’mRNA Library Prep Kit (Lexogen). Second strand of cDNA library was synthesized using the UMI Second Strand Synthesis Mix (USS, Lexogen) containing 6 nucleotides long Unique Molecular Identifiers (UMIs). The double stranded cDNA libraries were amplified in 17-19 PCR cycles using i5 Unique Dual Indexing Add-on Kit (Lexogen). Illumina NextSeq 500 was employed to sequence the libraries having a read lengths of single-end 75 bp producing about 10 million reads per library. To carryout gene expression analysis, Bioconductor R-package - DESeq2 v1.20.0 (Love et al., 2014) was used where in, featureCounts tool v1.6.3 (Liao et al., 2013) was used to enumerate the genes. A minimum threshold of adjusted p-value < 0.05 and log2fold-change (log2FC) = ± 1 was set to considered the gene as differentially expressed (DEG).

2023-10-31 | E-MTAB-13401 | biostudies-arrayexpress

Transcriptome analysis of 3D model of blood-brain barrier exposed to Borrelia garini

Project description:Using RNA sequencing to map differentially expressed genes in the 3D model of the blood-brain barrier ( composed of human brain endothelial cells, human astrocytes, and human pericytes) challenged with Borrelia garini .

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Ornithinibacillus bavariensis

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Neisseria meningitidis elicits a pro-inflammatory response involving IM-NM-:BM-NM-6 in a human blood-cerebrospinal fluid barrier model.

Project description:The human-specific, Gram-negative bacterium Neisseria meningitidis (Nm) is a leading cause of bacterial meningitis world-wide. It has been described that Nm can enter the central nervous system via the blood-cerebrospinal fluid barrier (BCSFB), which is constituted by the epithelial cells of the choroid plexus. Using a recently established in vitro model of the human BCSFB based on human malignant choroid plexus papilloma (HIBCPP) cells we investigated the cellular response of HIBCPP cells challenged with the meningitis-causing Nm strain MC58. In comparison we analysed the answer to the closely related unencapsulated carrier isolate Nm M-NM-114. Transcriptome analysis revealed a stronger transcriptional response after infection with strain MC58, in particular with its capsule deficient mutant MC58siaD-, which correlated with bacterial invasion levels. Expression evaluation and Gene Set Enrichment Analysis pointed to a NF-M-NM-:B-mediated pro-inflammatory immune response involving up-regulation of the transcription factor IM-NM-:BM-NM-6. Consistent with this, infected cells secreted significant levels of pro-inflammatory chemokines and cytokines, among others, IL8, CXCL1-3 and the IM-NM-:BM-NM-6 target gene product IL6. Expression profile of pattern recognition receptors in HIBCPP cells and the response to specific agonists indicates that TLR2 rather than TLR4 is involved in the cellular reaction following Nm infection. Human malignant choroid plexus papilloma (HIBCPP) cells were infected from the basolateral side with the meningitis-causing Neisseria meningitidis disease isolate MC58, its non-capsulated mutant MC58siaD- and the Neisseria meningitidis carrier isolate M-NM-114 for 4 h.The transcriptional response of HIBCPP cells to the different Neisseria meningitidis strains was evaluated by microarray analysis. Untreated HIBCPP cells served as control. Three replicates of each condition were analysed.

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The phosphoproteome of choroid plexus epithelial cells following infection with Neisseria meningitidis

Project description:The Gram-negative bacterium Neisseria meningitidis causes meningitis in humans and has been demonstrated to manipulate or alter host signalling pathways during infection of the central nervous system. In this study, the phosphoproteome of an in vitro model of the blood-cerebrospinal fluid barrier was investigated during infection with the Neisseria meningitidis serogroup B (NmB) strain MC58 in presence and absence of the bacterial capsule. We show that the capsule deficient mutant has a higher impact on the phosphoproteome of the infected cells and identify potentially regulated pathways and cellular processes during infection.

2023-03-31 | PXD038560 | Pride

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Zur regulon of Neisseria meningitidis

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