TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage

Objective: To further our understanding of the role of tumor necrosis factor (TNF)α on the inflammatory response in chondrocytes. Design: We explored the effects of TNFα on the transcriptome of epiphyseal chondrocytes from newborn C57BL/6 mice at the total and single cell (sc) resolution. Results: G...

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Main Authors: Ernesto Canalis, Lauren Schilling, Emily Denker
Format: Article
Language:English
Published: Elsevier 2024-12-01
Series:Osteoarthritis and Cartilage Open
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Online Access:http://www.sciencedirect.com/science/article/pii/S2665913124000955
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author Ernesto Canalis
Lauren Schilling
Emily Denker
author_facet Ernesto Canalis
Lauren Schilling
Emily Denker
author_sort Ernesto Canalis
collection DOAJ
description Objective: To further our understanding of the role of tumor necrosis factor (TNF)α on the inflammatory response in chondrocytes. Design: We explored the effects of TNFα on the transcriptome of epiphyseal chondrocytes from newborn C57BL/6 mice at the total and single cell (sc) resolution. Results: Gene set enrichment analysis of total RNA-Seq from TNFα-treated chondrocytes revealed enhanced response to biotic stimulus, defense and immune response and cytokine signaling and suppressed cartilage and skeletal morphogenesis and development. scRNA-Seq analyzed 14,239 ​cells and 24,320 genes and distinguished 16 ​cell clusters. The more prevalent ones were constituted by limb bud and chondrogenic cells and fibroblasts comprising ∼73 ​% of the cell population. Genes expressed by joint fibroblasts were detected in 5 clusters comprising ∼45 ​% of the cells isolated. Pseudotime trajectory finding revealed an association between fibroblast and chondrogenic clusters which was not modified by TNFα. TNFα decreased the total cells recovered by 18.5 ​% and the chondrogenic, limb bud and mesenchymal clusters by 32 ​%, 27 ​% and 7 ​%, respectively. TNFα had profound effects on the insulin-like growth factor (IGF) axis decreasing Igf1, Igf2 and Igfbp4 and inducing Igfbp3 and Igfbp5, explaining an inhibition of collagen biosynthesis, cartilage and skeletal morphogenesis. Ingenuity Pathway Analysis of scRNA-Seq data revealed that TNFα enhanced the osteoarthritis, rheumatoid arthritis, pathogen induced cytokine storm and interleukin 6 signaling pathways and suppressed fibroblast growth factor signaling. Conclusions: Epiphyseal chondrocytes are constituted by diverse cell populations distinctly regulated by TNFα to promote inflammation and suppression of matrix biosynthesis and growth.
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spelling doaj-art-106197f5527e4491bbeb6c41005a75012025-08-20T02:05:28ZengElsevierOsteoarthritis and Cartilage Open2665-91312024-12-016410052810.1016/j.ocarto.2024.100528TNFα has differential effects on the transcriptome profile of selected populations in murine cartilageErnesto Canalis0Lauren Schilling1Emily Denker2Departments of Orthopaedic Surgery, UConn Health, Farmington, CT 06030, USA; Departments of Medicine, UConn Health, Farmington, CT 06030, USA; UConn Musculoskeletal Institute, UConn Health, Farmington, CT 06030, USA; Corresponding author. Department of Orthopaedic Surgery, UConn Health, Farmington, CT 06030-4037, USA. Tel.: +860 679 7978; fax: +860 679 1474UConn Musculoskeletal Institute, UConn Health, Farmington, CT 06030, USAUConn Musculoskeletal Institute, UConn Health, Farmington, CT 06030, USAObjective: To further our understanding of the role of tumor necrosis factor (TNF)α on the inflammatory response in chondrocytes. Design: We explored the effects of TNFα on the transcriptome of epiphyseal chondrocytes from newborn C57BL/6 mice at the total and single cell (sc) resolution. Results: Gene set enrichment analysis of total RNA-Seq from TNFα-treated chondrocytes revealed enhanced response to biotic stimulus, defense and immune response and cytokine signaling and suppressed cartilage and skeletal morphogenesis and development. scRNA-Seq analyzed 14,239 ​cells and 24,320 genes and distinguished 16 ​cell clusters. The more prevalent ones were constituted by limb bud and chondrogenic cells and fibroblasts comprising ∼73 ​% of the cell population. Genes expressed by joint fibroblasts were detected in 5 clusters comprising ∼45 ​% of the cells isolated. Pseudotime trajectory finding revealed an association between fibroblast and chondrogenic clusters which was not modified by TNFα. TNFα decreased the total cells recovered by 18.5 ​% and the chondrogenic, limb bud and mesenchymal clusters by 32 ​%, 27 ​% and 7 ​%, respectively. TNFα had profound effects on the insulin-like growth factor (IGF) axis decreasing Igf1, Igf2 and Igfbp4 and inducing Igfbp3 and Igfbp5, explaining an inhibition of collagen biosynthesis, cartilage and skeletal morphogenesis. Ingenuity Pathway Analysis of scRNA-Seq data revealed that TNFα enhanced the osteoarthritis, rheumatoid arthritis, pathogen induced cytokine storm and interleukin 6 signaling pathways and suppressed fibroblast growth factor signaling. Conclusions: Epiphyseal chondrocytes are constituted by diverse cell populations distinctly regulated by TNFα to promote inflammation and suppression of matrix biosynthesis and growth.http://www.sciencedirect.com/science/article/pii/S2665913124000955TNFαChondrocytesInflammationTranscriptomeIGF1IGF2
spellingShingle Ernesto Canalis
Lauren Schilling
Emily Denker
TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
Osteoarthritis and Cartilage Open
TNFα
Chondrocytes
Inflammation
Transcriptome
IGF1
IGF2
title TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
title_full TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
title_fullStr TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
title_full_unstemmed TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
title_short TNFα has differential effects on the transcriptome profile of selected populations in murine cartilage
title_sort tnfα has differential effects on the transcriptome profile of selected populations in murine cartilage
topic TNFα
Chondrocytes
Inflammation
Transcriptome
IGF1
IGF2
url http://www.sciencedirect.com/science/article/pii/S2665913124000955
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