Dual RNASeq reveals NTHi-macrophage transcriptomic changes during intracellular persistence
Dual RNASeq reveals NTHi-macrophage transcriptomic changes during intracellular persistence
Nontypeable Haemophilus influenzae (NTHi) is a pathobiont which chronically colonises the airway of individuals with chronic respiratory disease and is associated with poor clinical outcomes. It is unclear how NTHi persists in the airway, however accumulating evidence suggests that NTHi can invade and persist within macrophages. To better understand the mechanisms of NTHi persistence within macrophages, we developed an in vitro model of NTHi intracellular persistence using human monocyte-derived macrophages (MDM). Dual RNA Sequencing was used to assess MDM and NTHi transcriptomic regulation occurring simultaneously during NTHi persistence. Analysis of the macrophage response to NTHi identified temporally regulated transcriptomic profiles, with a specific ‘core’ profile displaying conserved expression of genes across time points. Gene list enrichment analysis identified enrichment of immune responses in the core gene set, with KEGG pathway analysis revealing
specific enrichment of intracellular immune response pathways. NTHi persistence was facilitated by modulation of bacterial metabolic, stress response and ribosome pathways. Levels of NTHi genes bioC, mepM and dps were differentially expressed by intracellular NTHi compared to planktonic NTHi, indicating that the transcriptomic adaption was distinct between the two different NTHi lifestyles. Overall, this study provides crucial insights into the transcriptomic adaptations facilitating NTHi persistence within macrophages. Targeting these reported pathways with novel therapeutics to reduce NTHi burden in the airway could be an effective treatment strategy given the current antimicrobial resistance crisis and lack of NTHi vaccines.
NTHi, dual RNAseq, host-pathogen interactions, intracellular persistence, macrophage
Ackland, Jodie
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Heinson, Ashley I.
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Cleary, David W.
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Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Wilkinson, Tom M
8c55ebbb-e547-445c-95a1-c8bed02dd652
Staples, Karl J.
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23 August 2021
Ackland, Jodie
dba59510-7535-47f8-b2ba-2d49dfa7fbd8
Heinson, Ashley I.
822775d1-9379-4bde-99c3-3c031c3100fb
Cleary, David W.
f4079c6d-d54b-4108-b346-b0069035bec0
Christodoulides, Myron
eba99148-620c-452a-a334-c1a52ba94078
Wilkinson, Tom M
8c55ebbb-e547-445c-95a1-c8bed02dd652
Staples, Karl J.
e0e9d80f-0aed-435f-bd75-0c8818491fee
Ackland, Jodie, Heinson, Ashley I., Cleary, David W., Christodoulides, Myron, Wilkinson, Tom M and Staples, Karl J.
(2021)
Dual RNASeq reveals NTHi-macrophage transcriptomic changes during intracellular persistence.
Frontiers in Cellular and Infection Microbiology, 11, [723481].
(doi:10.3389/fcimb.2021.723481).
Abstract
Nontypeable Haemophilus influenzae (NTHi) is a pathobiont which chronically colonises the airway of individuals with chronic respiratory disease and is associated with poor clinical outcomes. It is unclear how NTHi persists in the airway, however accumulating evidence suggests that NTHi can invade and persist within macrophages. To better understand the mechanisms of NTHi persistence within macrophages, we developed an in vitro model of NTHi intracellular persistence using human monocyte-derived macrophages (MDM). Dual RNA Sequencing was used to assess MDM and NTHi transcriptomic regulation occurring simultaneously during NTHi persistence. Analysis of the macrophage response to NTHi identified temporally regulated transcriptomic profiles, with a specific ‘core’ profile displaying conserved expression of genes across time points. Gene list enrichment analysis identified enrichment of immune responses in the core gene set, with KEGG pathway analysis revealing
specific enrichment of intracellular immune response pathways. NTHi persistence was facilitated by modulation of bacterial metabolic, stress response and ribosome pathways. Levels of NTHi genes bioC, mepM and dps were differentially expressed by intracellular NTHi compared to planktonic NTHi, indicating that the transcriptomic adaption was distinct between the two different NTHi lifestyles. Overall, this study provides crucial insights into the transcriptomic adaptations facilitating NTHi persistence within macrophages. Targeting these reported pathways with novel therapeutics to reduce NTHi burden in the airway could be an effective treatment strategy given the current antimicrobial resistance crisis and lack of NTHi vaccines.
Other
Ackland et al 2021 Front Cell Infect Micro Accepted
- Accepted Manuscript
More information
Accepted/In Press date: 26 July 2021
Published date: 23 August 2021
Additional Information:
Funding Information:
The authors would like to express gratitude to all the volunteers who kindly provided blood samples for this study. The authors also wish to thank Richard Jewell and Dr Carolann McGuire of the University of Southampton, Faculty of Medicine Flow Cytometry Unit and Dr Alastair Watson for his help in preparing the manuscript for submission.
Funding Information:
This work was primarily funded by an Asthma UK studentship award (AUK-PHD-2016-363). We also gratefully acknowledge the support of the Southampton AAIR charity in funding this work. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© Copyright © 2021 Ackland, Heinson, Cleary, Christodoulides, Wilkinson and Staples.
Keywords:
NTHi, dual RNAseq, host-pathogen interactions, intracellular persistence, macrophage
Identifiers
Local EPrints ID: 450536
URI: http://eprints.soton.ac.uk/id/eprint/450536
ISSN: 2235-2988
PURE UUID: 80cd6e73-e33b-4142-ae18-3e9c3e8102ea
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Date deposited: 03 Aug 2021 16:31
Last modified: 25 Apr 2024 17:37
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Author:
Ashley I. Heinson
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