PUBLICATION

Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum

Authors

Kenyon, A., Gavriouchkina, D., Zorman, J., Napolitani, G., Cerundolo, V., Sauka-Spengler, T.

ID

ZDB-PUB-170728-5

Date

2017

Source

Scientific Reports 7: 6505 (Journal)

Registered Authors

Sauka-Spengler, Tatjana

Keywords

Neutrophils, Transcriptomics

Datasets

GEO:GSE92740

MeSH Terms

Animals
Sequence Analysis, RNA
Mycobacterium marinum/immunology*
Inflammasomes/metabolism*
Immunity, Innate*
Disease Models, Animal
Zebrafish
Mycobacterium Infections, Nontuberculous/pathology*
Gene Expression Profiling*
Neutrophils/immunology*

(all 10)

PubMed

28747644 Full text @ Sci. Rep.

Abstract

The mechanisms governing neutrophil response to Mycobacterium tuberculosis remain poorly understood. In this study we utilise biotagging, a novel genome-wide profiling approach based on cell type-specific in vivo biotinylation in zebrafish to analyse the initial response of neutrophils to Mycobacterium marinum, a close genetic relative of M. tuberculosis used to model tuberculosis. Differential expression analysis following nuclear RNA-seq of neutrophil active transcriptomes reveals a significant upregulation in both damage-sensing and effector components of the inflammasome, including caspase b, NLRC3 ortholog (wu: fb15h11) and il1β. Crispr/Cas9-mediated knockout of caspase b, which acts by proteolytic processing of il1β, results in increased bacterial burden and less infiltration of macrophages to sites of mycobacterial infection, thus impairing granuloma development. We also show that a number of immediate early response genes (IEGs) are responsible for orchestrating the initial neutrophil response to mycobacterial infection. Further perturbation of the IEGs exposes egr3 as a key transcriptional regulator controlling il1β transcription.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
WT	bacterial treatment by injection: Mycobacterium marinum	Long-pec to Day 5	neutrophil increased amount, abnormal	Fig. 1
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	myeloid cell caspb expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	myeloid cell egr3 expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	myeloid cell nlrp16 expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism cebpb expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism cxcl8b.1 expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism fosl1a expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism fosl1a expression increased distribution, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism hbegfa expression increased amount, abnormal	Fig. 3
WT	bacterial treatment by injection: Mycobacterium marinum	Protruding-mouth	whole organism hbegfa expression increased distribution, abnormal	Fig. 3

1 - 10 of 33

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Mutations / Transgenics

Allele	Construct	Type
ct700aTg	Tg(actb2:AVI-Cerulean-Gga.Rangap1)	Transgenic Insertion
gl22Tg	Tg(mpeg1:EGFP)	Transgenic Insertion
ox121Tg	TgBAC(mpx:Eco.BirA-Citrine)	Transgenic Insertion

1 - 3 of 3

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Human Disease / Model

Human Disease	Fish	Conditions	Evidence
bacterial infectious disease	WT	bacterial treatment by injection: Mycobacterium marinum	TAS
tuberculosis	WT	bacterial treatment by injection: Mycobacterium marinum	TAS

1 - 2 of 2

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Sequence Targeting Reagents

Target	Reagent	Reagent Type
caspb	CRISPR1-caspb	CRISPR
caspb	CRISPR2-caspb	CRISPR
caspb	CRISPR3-caspb	CRISPR
egr3	CRISPR1-egr3	CRISPR
egr3	CRISPR2-egr3	CRISPR
egr3	CRISPR3-egr3	CRISPR
junbb	CRISPR4-junbb	CRISPR
junbb	CRISPR5-junbb	CRISPR
junbb	CRISPR6-junbb	CRISPR
nlrc6	CRISPR1-nlrp16	CRISPR

1 - 10 of 20

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Fish

Fish
ct700aTg; ox121Tg
gl22Tg
gl22Tg + CRISPR1-caspb
gl22Tg + CRISPR2-caspb
gl22Tg + CRISPR3-caspb
WT
WT + CRISPR1-egr3
WT + CRISPR2-egr3
WT + CRISPR3-egr3

Antibodies

Name	Type	Antigen Genes	Isotypes	Host Organism
Ab4-mpx	polyclonal		IgG	Rabbit

Orthology

Engineered Foreign Genes

Marker	Marker Type	Name
Cerulean	EFG	Cerulean
Citrine	EFG	Citrine
EGFP	EFG	EGFP

Mapping

Kenyon et al., 2017 ZDB-PUB-170728-5 PMID:28747644

Active nuclear transcriptome analysis reveals inflammasome-dependent mechanism for early neutrophil response to Mycobacterium marinum

Kenyon et al., 2017

ZDB-PUB-170728-5
PMID:28747644