Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets

Dominika E. Nanus; Amel Badoume; Susanne N. Wijesinghe; Andrea M. Halsey; Patrick Hurley; Zubair Ahmed; Rajesh Botchu; Edward T. Davis; Mark A. Lindsay; Simon W. Jones

doi:10.1016/j.ebiom.2021.103618

Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets

Dominika E. Nanus, Amel Badoume, Susanne N. Wijesinghe, Andrea M. Halsey, Patrick Hurley, Zubair Ahmed, Rajesh Botchu, Edward T. Davis, Mark A. Lindsay, Simon W. Jones

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

56 Citations (SciVal)

Abstract

Background: Synovial inflammation is associated with pain severity in patients with knee osteoarthritis (OA). The aim here was to determine in a population with knee OA, whether synovial tissue from areas associated with pain exhibited different synovial fibroblast subsets, compared to synovial tissue from sites not associated with pain. A further aim was to compare differences between early and end-stage disease synovial fibroblast subsets. Methods: Patients with early knee OA (n = 29) and end-stage knee OA (n = 22) were recruited. Patient reported pain was recorded by questionnaire and using an anatomical knee pain map. Proton density fat suppressed MRI axial and sagittal sequences were analysed and scored for synovitis. Synovial tissue was obtained from the medial and lateral parapatellar and suprapatellar sites. Fibroblast single cell RNA sequencing was performed using Chromium 10X and analysed using Seurat. Transcriptomes were functionally characterised using Ingenuity Pathway Analysis and the effect of fibroblast secretome on neuronal growth assessed using rat DRGN. Findings: Parapatellar synovitis was significantly associated with the pattern of patient-reported pain in knee OA patients. Synovial tissue from sites of patient-reported pain exhibited a differential transcriptomic phenotype, with distinct synovial fibroblast subsets in early OA and end-stage OA. Functional pathway analysis revealed that synovial tissue and fibroblast subsets from painful sites promoted fibrosis, inflammation and the growth and activity of neurons. The secretome of fibroblasts from early OA painful sites induced greater survival and neurite outgrowth in dissociated adult rodent dorsal root ganglion neurons. Interpretation: Sites of patient-reported pain in knee OA exhibit a different synovial tissue phenotype and distinct synovial fibroblast subsets. Further interrogation of these fibroblast pathotypes will increase our understanding of the role of synovitis in OA joint pain and provide a rationale for the therapeutic targeting of fibroblast subsets to alleviate pain in patients. Funding: This study was funded by Versus Arthritis, UK (21530; 21812)

Original language	English
Article number	103618
Journal	EBioMedicine
Volume	72
Early online date	7 Oct 2021
DOIs	https://doi.org/10.1016/j.ebiom.2021.103618
Publication status	Published - 31 Oct 2021

Bibliographical note

Funding Information:
This study was funded by Versus Arthritis (21530; 21812). The funder had no involvement in the study or any role in the writing of the manuscript or the decision to submit it for publication. The corresponding author (Dr Simon Jones) confirms that authors were not precluded from accessing data in the study and they accept responsibility to submit for publication. The authors acknowledge all study participants, research staff at The Royal Orthopaedic Hospital NHS Foundation Trust and Russell's Hall Hospital, Dudley for obtaining consents and screening. This research made use of the Balena High Performance Computing (HPC) Service at the University of Bath.

Funding Information:
SWJ declares grant funding from Versus Arthritis during the course of this study.

Data sharing statement The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE176223 and GSE176308.

Keywords

Inflammation
Obesity
Osteoarthritis
scRNAseq
Synovial fibroblasts

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1016/j.ebiom.2021.103618Licence: CC BY-NC-ND

Cite this

@article{d395575816454f45a2ae2c24624ae488,

title = "Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets",

abstract = "Background: Synovial inflammation is associated with pain severity in patients with knee osteoarthritis (OA). The aim here was to determine in a population with knee OA, whether synovial tissue from areas associated with pain exhibited different synovial fibroblast subsets, compared to synovial tissue from sites not associated with pain. A further aim was to compare differences between early and end-stage disease synovial fibroblast subsets. Methods: Patients with early knee OA (n = 29) and end-stage knee OA (n = 22) were recruited. Patient reported pain was recorded by questionnaire and using an anatomical knee pain map. Proton density fat suppressed MRI axial and sagittal sequences were analysed and scored for synovitis. Synovial tissue was obtained from the medial and lateral parapatellar and suprapatellar sites. Fibroblast single cell RNA sequencing was performed using Chromium 10X and analysed using Seurat. Transcriptomes were functionally characterised using Ingenuity Pathway Analysis and the effect of fibroblast secretome on neuronal growth assessed using rat DRGN. Findings: Parapatellar synovitis was significantly associated with the pattern of patient-reported pain in knee OA patients. Synovial tissue from sites of patient-reported pain exhibited a differential transcriptomic phenotype, with distinct synovial fibroblast subsets in early OA and end-stage OA. Functional pathway analysis revealed that synovial tissue and fibroblast subsets from painful sites promoted fibrosis, inflammation and the growth and activity of neurons. The secretome of fibroblasts from early OA painful sites induced greater survival and neurite outgrowth in dissociated adult rodent dorsal root ganglion neurons. Interpretation: Sites of patient-reported pain in knee OA exhibit a different synovial tissue phenotype and distinct synovial fibroblast subsets. Further interrogation of these fibroblast pathotypes will increase our understanding of the role of synovitis in OA joint pain and provide a rationale for the therapeutic targeting of fibroblast subsets to alleviate pain in patients. Funding: This study was funded by Versus Arthritis, UK (21530; 21812)",

keywords = "Inflammation, Obesity, Osteoarthritis, scRNAseq, Synovial fibroblasts",

author = "Nanus, {Dominika E.} and Amel Badoume and Wijesinghe, {Susanne N.} and Halsey, {Andrea M.} and Patrick Hurley and Zubair Ahmed and Rajesh Botchu and Davis, {Edward T.} and Lindsay, {Mark A.} and Jones, {Simon W.}",

note = "Funding Information: This study was funded by Versus Arthritis (21530; 21812). The funder had no involvement in the study or any role in the writing of the manuscript or the decision to submit it for publication. The corresponding author (Dr Simon Jones) confirms that authors were not precluded from accessing data in the study and they accept responsibility to submit for publication. The authors acknowledge all study participants, research staff at The Royal Orthopaedic Hospital NHS Foundation Trust and Russell's Hall Hospital, Dudley for obtaining consents and screening. This research made use of the Balena High Performance Computing (HPC) Service at the University of Bath. Funding Information: SWJ declares grant funding from Versus Arthritis during the course of this study. Data sharing statement The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE176223 and GSE176308.",

year = "2021",

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day = "31",

doi = "10.1016/j.ebiom.2021.103618",

language = "English",

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journal = "EBioMedicine",

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T1 - Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets

AU - Nanus, Dominika E.

AU - Badoume, Amel

AU - Wijesinghe, Susanne N.

AU - Halsey, Andrea M.

AU - Hurley, Patrick

AU - Ahmed, Zubair

AU - Botchu, Rajesh

AU - Davis, Edward T.

AU - Lindsay, Mark A.

AU - Jones, Simon W.

N1 - Funding Information: This study was funded by Versus Arthritis (21530; 21812). The funder had no involvement in the study or any role in the writing of the manuscript or the decision to submit it for publication. The corresponding author (Dr Simon Jones) confirms that authors were not precluded from accessing data in the study and they accept responsibility to submit for publication. The authors acknowledge all study participants, research staff at The Royal Orthopaedic Hospital NHS Foundation Trust and Russell's Hall Hospital, Dudley for obtaining consents and screening. This research made use of the Balena High Performance Computing (HPC) Service at the University of Bath. Funding Information: SWJ declares grant funding from Versus Arthritis during the course of this study. Data sharing statement The data discussed in this publication have been deposited in NCBI's Gene Expression Omnibus and are accessible through GEO Series accession number GSE176223 and GSE176308.

PY - 2021/10/31

Y1 - 2021/10/31

N2 - Background: Synovial inflammation is associated with pain severity in patients with knee osteoarthritis (OA). The aim here was to determine in a population with knee OA, whether synovial tissue from areas associated with pain exhibited different synovial fibroblast subsets, compared to synovial tissue from sites not associated with pain. A further aim was to compare differences between early and end-stage disease synovial fibroblast subsets. Methods: Patients with early knee OA (n = 29) and end-stage knee OA (n = 22) were recruited. Patient reported pain was recorded by questionnaire and using an anatomical knee pain map. Proton density fat suppressed MRI axial and sagittal sequences were analysed and scored for synovitis. Synovial tissue was obtained from the medial and lateral parapatellar and suprapatellar sites. Fibroblast single cell RNA sequencing was performed using Chromium 10X and analysed using Seurat. Transcriptomes were functionally characterised using Ingenuity Pathway Analysis and the effect of fibroblast secretome on neuronal growth assessed using rat DRGN. Findings: Parapatellar synovitis was significantly associated with the pattern of patient-reported pain in knee OA patients. Synovial tissue from sites of patient-reported pain exhibited a differential transcriptomic phenotype, with distinct synovial fibroblast subsets in early OA and end-stage OA. Functional pathway analysis revealed that synovial tissue and fibroblast subsets from painful sites promoted fibrosis, inflammation and the growth and activity of neurons. The secretome of fibroblasts from early OA painful sites induced greater survival and neurite outgrowth in dissociated adult rodent dorsal root ganglion neurons. Interpretation: Sites of patient-reported pain in knee OA exhibit a different synovial tissue phenotype and distinct synovial fibroblast subsets. Further interrogation of these fibroblast pathotypes will increase our understanding of the role of synovitis in OA joint pain and provide a rationale for the therapeutic targeting of fibroblast subsets to alleviate pain in patients. Funding: This study was funded by Versus Arthritis, UK (21530; 21812)

AB - Background: Synovial inflammation is associated with pain severity in patients with knee osteoarthritis (OA). The aim here was to determine in a population with knee OA, whether synovial tissue from areas associated with pain exhibited different synovial fibroblast subsets, compared to synovial tissue from sites not associated with pain. A further aim was to compare differences between early and end-stage disease synovial fibroblast subsets. Methods: Patients with early knee OA (n = 29) and end-stage knee OA (n = 22) were recruited. Patient reported pain was recorded by questionnaire and using an anatomical knee pain map. Proton density fat suppressed MRI axial and sagittal sequences were analysed and scored for synovitis. Synovial tissue was obtained from the medial and lateral parapatellar and suprapatellar sites. Fibroblast single cell RNA sequencing was performed using Chromium 10X and analysed using Seurat. Transcriptomes were functionally characterised using Ingenuity Pathway Analysis and the effect of fibroblast secretome on neuronal growth assessed using rat DRGN. Findings: Parapatellar synovitis was significantly associated with the pattern of patient-reported pain in knee OA patients. Synovial tissue from sites of patient-reported pain exhibited a differential transcriptomic phenotype, with distinct synovial fibroblast subsets in early OA and end-stage OA. Functional pathway analysis revealed that synovial tissue and fibroblast subsets from painful sites promoted fibrosis, inflammation and the growth and activity of neurons. The secretome of fibroblasts from early OA painful sites induced greater survival and neurite outgrowth in dissociated adult rodent dorsal root ganglion neurons. Interpretation: Sites of patient-reported pain in knee OA exhibit a different synovial tissue phenotype and distinct synovial fibroblast subsets. Further interrogation of these fibroblast pathotypes will increase our understanding of the role of synovitis in OA joint pain and provide a rationale for the therapeutic targeting of fibroblast subsets to alleviate pain in patients. Funding: This study was funded by Versus Arthritis, UK (21530; 21812)

KW - Inflammation

KW - Obesity

KW - Osteoarthritis

KW - scRNAseq

KW - Synovial fibroblasts

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U2 - 10.1016/j.ebiom.2021.103618

DO - 10.1016/j.ebiom.2021.103618

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Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Light-sheet microscope Zeiss Z.1

Cite this

Synovial tissue from sites of joint pain in knee osteoarthritis patients exhibits a differential phenotype with distinct fibroblast subsets

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

Access to Document

Other files and links

Fingerprint

Equipment

Light-sheet microscope Zeiss Z.1

Cite this