Single-cell RNA sequencing analysis of vestibular schwannoma reveals functionally distinct macrophage subsets

Paramita Baruah; Christopher Mahony; Jennifer L. Marshall; Charlotte G. Smith; Peter Monksfield; Richard I. Irving; Ingrid E. Dumitriu; Christopher D. Buckley; Adam P. Croft

doi:10.1038/s41416-024-02646-2

Single-cell RNA sequencing analysis of vestibular schwannoma reveals functionally distinct macrophage subsets

Paramita Baruah^*, Christopher Mahony, Jennifer L. Marshall, Charlotte G. Smith, Peter Monksfield, Richard I. Irving, Ingrid E. Dumitriu, Christopher D. Buckley, Adam P. Croft

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

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Abstract

BACKGROUND: Vestibular schwannomas (VSs) remain a challenge due to their anatomical location and propensity to growth. Macrophages are present in VS but their roles in VS pathogenesis remains unknown.

OBJECTIVES: The objective was to assess phenotypic and functional profile of macrophages in VS with single-cell RNA sequencing (scRNAseq).

METHODS: scRNAseq was carried out in three VS samples to examine characteristics of macrophages in the tumour. RT-qPCR was carried out on 10 VS samples for CD14, CD68 and CD163 and a panel of macrophage-associated molecules.

RESULTS: scRNAseq revealed macrophages to be a major constituent of VS microenvironment with three distinct subclusters based on gene expression. The subclusters were also defined by expression of CD163, CD68 and IL-1β. AREG and PLAUR were expressed in the CD68+CD163+IL-1β+ subcluster, PLCG2 and NCKAP5 were expressed in CD68+CD163+IL-1β- subcluster and AUTS2 and SPP1 were expressed in the CD68+CD163-IL-1β+ subcluster. RT-qPCR showed expression of several macrophage markers in VS of which CD14, ALOX15, Interleukin-1β, INHBA and Colony Stimulating Factor-1R were found to have a high correlation with tumour volume.

CONCLUSIONS: Macrophages form an important component of VS stroma. scRNAseq reveals three distinct subsets of macrophages in the VS tissue which may have differing roles in the pathogenesis of VS.

Original language	English
Journal	British Journal of Cancer
Early online date	13 Mar 2024
DOIs	https://doi.org/10.1038/s41416-024-02646-2
Publication status	E-pub ahead of print - 13 Mar 2024

Bibliographical note

Funding:
This report includes independent research supported by the National Institute for Health Research through the Birmingham Biomedical Research Center and Wellcome Trust Clinical Research Facility at University Hospitals Birmingham NHS Foundation Trust. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, our funding bodies or the Department of Health. Funding was also provided by the Versus Arthritis RACE Rheumatoid Arthritis Pathogenesis Centre of Excellence (grant20298) and a Versus Arthritis grant to CDB (grant 19791). Funding was provided by a Royal College of Surgeons Grant and Midlands Institute of Otology grant to PB. Funding was provided by Versus Arthritis, NIHR and Wellcome Trust to AC. The funding organisations did not have any role in study design, data collection, analysis and interpretation, manuscript writing and decision to submit the article for publication.

Copyright:
© 2024. The Author(s).

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BaruahP2024SingleFinal published version, 1.94 MBLicence: Creative Commons: Attribution (CC BY)

Targetting stromal cell subsets in the pathogenesis of imflammatory Arthritis
Buckley, C.
ARTHRITIS RESEARCH CAMPAIGN
1/05/12 → 31/10/18
Project: Research

Cite this

@article{eacd0d5175ca473eb98158ec9b7db7d4,

title = "Single-cell RNA sequencing analysis of vestibular schwannoma reveals functionally distinct macrophage subsets",

abstract = "BACKGROUND: Vestibular schwannomas (VSs) remain a challenge due to their anatomical location and propensity to growth. Macrophages are present in VS but their roles in VS pathogenesis remains unknown.OBJECTIVES: The objective was to assess phenotypic and functional profile of macrophages in VS with single-cell RNA sequencing (scRNAseq).METHODS: scRNAseq was carried out in three VS samples to examine characteristics of macrophages in the tumour. RT-qPCR was carried out on 10 VS samples for CD14, CD68 and CD163 and a panel of macrophage-associated molecules.RESULTS: scRNAseq revealed macrophages to be a major constituent of VS microenvironment with three distinct subclusters based on gene expression. The subclusters were also defined by expression of CD163, CD68 and IL-1β. AREG and PLAUR were expressed in the CD68+CD163+IL-1β+ subcluster, PLCG2 and NCKAP5 were expressed in CD68+CD163+IL-1β- subcluster and AUTS2 and SPP1 were expressed in the CD68+CD163-IL-1β+ subcluster. RT-qPCR showed expression of several macrophage markers in VS of which CD14, ALOX15, Interleukin-1β, INHBA and Colony Stimulating Factor-1R were found to have a high correlation with tumour volume.CONCLUSIONS: Macrophages form an important component of VS stroma. scRNAseq reveals three distinct subsets of macrophages in the VS tissue which may have differing roles in the pathogenesis of VS.",

author = "Paramita Baruah and Christopher Mahony and Marshall, {Jennifer L.} and Smith, {Charlotte G.} and Peter Monksfield and Irving, {Richard I.} and Dumitriu, {Ingrid E.} and Buckley, {Christopher D.} and Croft, {Adam P.}",

note = "Funding: This report includes independent research supported by the National Institute for Health Research through the Birmingham Biomedical Research Center and Wellcome Trust Clinical Research Facility at University Hospitals Birmingham NHS Foundation Trust. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, our funding bodies or the Department of Health. Funding was also provided by the Versus Arthritis RACE Rheumatoid Arthritis Pathogenesis Centre of Excellence (grant20298) and a Versus Arthritis grant to CDB (grant 19791). Funding was provided by a Royal College of Surgeons Grant and Midlands Institute of Otology grant to PB. Funding was provided by Versus Arthritis, NIHR and Wellcome Trust to AC. The funding organisations did not have any role in study design, data collection, analysis and interpretation, manuscript writing and decision to submit the article for publication. Copyright: {\textcopyright} 2024. The Author(s).",

year = "2024",

month = mar,

day = "13",

doi = "10.1038/s41416-024-02646-2",

language = "English",

journal = "British Journal of Cancer",

issn = "0007-0920",

publisher = "Cancer Research UK",

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TY - JOUR

T1 - Single-cell RNA sequencing analysis of vestibular schwannoma reveals functionally distinct macrophage subsets

AU - Baruah, Paramita

AU - Mahony, Christopher

AU - Marshall, Jennifer L.

AU - Smith, Charlotte G.

AU - Monksfield, Peter

AU - Irving, Richard I.

AU - Dumitriu, Ingrid E.

AU - Buckley, Christopher D.

AU - Croft, Adam P.

N1 - Funding: This report includes independent research supported by the National Institute for Health Research through the Birmingham Biomedical Research Center and Wellcome Trust Clinical Research Facility at University Hospitals Birmingham NHS Foundation Trust. The views expressed are those of the author(s) and not necessarily those of the NHS, the NIHR, our funding bodies or the Department of Health. Funding was also provided by the Versus Arthritis RACE Rheumatoid Arthritis Pathogenesis Centre of Excellence (grant20298) and a Versus Arthritis grant to CDB (grant 19791). Funding was provided by a Royal College of Surgeons Grant and Midlands Institute of Otology grant to PB. Funding was provided by Versus Arthritis, NIHR and Wellcome Trust to AC. The funding organisations did not have any role in study design, data collection, analysis and interpretation, manuscript writing and decision to submit the article for publication. Copyright: © 2024. The Author(s).

PY - 2024/3/13

Y1 - 2024/3/13

N2 - BACKGROUND: Vestibular schwannomas (VSs) remain a challenge due to their anatomical location and propensity to growth. Macrophages are present in VS but their roles in VS pathogenesis remains unknown.OBJECTIVES: The objective was to assess phenotypic and functional profile of macrophages in VS with single-cell RNA sequencing (scRNAseq).METHODS: scRNAseq was carried out in three VS samples to examine characteristics of macrophages in the tumour. RT-qPCR was carried out on 10 VS samples for CD14, CD68 and CD163 and a panel of macrophage-associated molecules.RESULTS: scRNAseq revealed macrophages to be a major constituent of VS microenvironment with three distinct subclusters based on gene expression. The subclusters were also defined by expression of CD163, CD68 and IL-1β. AREG and PLAUR were expressed in the CD68+CD163+IL-1β+ subcluster, PLCG2 and NCKAP5 were expressed in CD68+CD163+IL-1β- subcluster and AUTS2 and SPP1 were expressed in the CD68+CD163-IL-1β+ subcluster. RT-qPCR showed expression of several macrophage markers in VS of which CD14, ALOX15, Interleukin-1β, INHBA and Colony Stimulating Factor-1R were found to have a high correlation with tumour volume.CONCLUSIONS: Macrophages form an important component of VS stroma. scRNAseq reveals three distinct subsets of macrophages in the VS tissue which may have differing roles in the pathogenesis of VS.

AB - BACKGROUND: Vestibular schwannomas (VSs) remain a challenge due to their anatomical location and propensity to growth. Macrophages are present in VS but their roles in VS pathogenesis remains unknown.OBJECTIVES: The objective was to assess phenotypic and functional profile of macrophages in VS with single-cell RNA sequencing (scRNAseq).METHODS: scRNAseq was carried out in three VS samples to examine characteristics of macrophages in the tumour. RT-qPCR was carried out on 10 VS samples for CD14, CD68 and CD163 and a panel of macrophage-associated molecules.RESULTS: scRNAseq revealed macrophages to be a major constituent of VS microenvironment with three distinct subclusters based on gene expression. The subclusters were also defined by expression of CD163, CD68 and IL-1β. AREG and PLAUR were expressed in the CD68+CD163+IL-1β+ subcluster, PLCG2 and NCKAP5 were expressed in CD68+CD163+IL-1β- subcluster and AUTS2 and SPP1 were expressed in the CD68+CD163-IL-1β+ subcluster. RT-qPCR showed expression of several macrophage markers in VS of which CD14, ALOX15, Interleukin-1β, INHBA and Colony Stimulating Factor-1R were found to have a high correlation with tumour volume.CONCLUSIONS: Macrophages form an important component of VS stroma. scRNAseq reveals three distinct subsets of macrophages in the VS tissue which may have differing roles in the pathogenesis of VS.

U2 - 10.1038/s41416-024-02646-2

DO - 10.1038/s41416-024-02646-2

M3 - Article

C2 - 38480935

SN - 0007-0920

JO - British Journal of Cancer

JF - British Journal of Cancer

ER -

Single-cell RNA sequencing analysis of vestibular schwannoma reveals functionally distinct macrophage subsets

Abstract

Bibliographical note

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Targetting stromal cell subsets in the pathogenesis of imflammatory Arthritis

Cite this