Expression and Activity of TRPA1 and TRPV1 in the Intervertebral Disc: Association with Inflammation and Matrix Remodeling

Takuya Kameda, Joel Zvick, Miriam Vuk, Aleksandra Sadowska, Wai Kit Tam, Victor Y. Leung, K. Bölcskei, Z. Helyes, Lee Ann Applegate, Oliver N. Hausmann, Juergen Klasen, Olga Krupkova, Karin Wuertz-Kozak

Research output: Contribution to journalArticle

Abstract

Transient receptor potential (TRP) channels have emerged as potential sensors and transducers of inflammatory pain. The aims of this study were to investigate (1) the expression of TRP channels in intervertebral disc (IVD) cells in normal and inflammatory conditions and (2) the function of Transient receptor potential ankyrin 1 (TRPA1) and Transient receptor potential vanilloid 1 (TRPV1) in IVD inflammation and matrix homeostasis. RT-qPCR was used to analyze human fetal, healthy, and degenerated IVD tissues for the gene expression of TRPA1 and TRPV1. The primary IVD cell cultures were stimulated with either interleukin-1 beta (IL-1β) or tumor necrosis factor alpha (TNF-α) alone or in combination with TRPA1/V1 agonist allyl isothiocyanate (AITC, 3 and 10 µM), followed by analysis of calcium flux and the expression of inflammation mediators (RT-qPCR/ELISA) and matrix constituents (RT-qPCR). The matrix structure and composition in caudal motion segments from TRPA1 and TRPV1 wild-type (WT) and knock-out (KO) mice was visualized by FAST staining. Gene expression of other TRP channels (A1, C1, C3, C6, V1, V2, V4, V6, M2, M7, M8) was also tested in cytokine-treated cells. TRPA1 was expressed in fetal IVD cells, 20% of degenerated IVDs, but not in healthy mature IVDs. TRPA1 expression was not detectable in untreated cells and it increased upon cytokine treatment, while TRPV1 was expressed and concomitantly reduced. In inflamed IVD cells, 10 µM AITC activated calcium flux, induced gene expression of IL-8, and reduced disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and collagen 1A1, possibly via upregulated TRPA1. TRPA1 KO in mice was associated with signs of degeneration in the nucleus pulposus and the vertebral growth plate, whereas TRPV1 KO did not show profound changes. Cytokine treatment also affected the gene expression of TRPV2 (increase), TRPV4 (increase), and TRPC6 (decrease). TRPA1 might be expressed in developing IVD, downregulated during its maturation, and upregulated again in degenerative disc disease, participating in matrix homeostasis. However, follow-up studies with larger sample sizes are needed to fully elucidate the role of TRPA1 and other TRP channels in degenerative disc disease.

Original languageEnglish
JournalInternational journal of molecular sciences
Volume20
Issue number7
DOIs
Publication statusPublished - Apr 10 2019

Fingerprint

intervertebral disks
Ankyrins
Intervertebral Disc
Inflammation
Transient Receptor Potential Channels
matrices
Gene Expression
Gene expression
gene expression
Cytokines
Knockout Mice
Homeostasis
Cartilage Oligomeric Matrix Protein
vanilloid receptor subtype 1
Calcium
Disintegrins
homeostasis
Inflammation Mediators
cells
Growth Plate

Keywords

  • aggrecanases
  • collagen
  • low back pain
  • pro-inflammatory cytokines
  • TRP channels
  • TRPA1
  • TRPC6
  • TRPV1
  • TRPV2
  • TRPV4

ASJC Scopus subject areas

  • Catalysis
  • Molecular Biology
  • Spectroscopy
  • Computer Science Applications
  • Physical and Theoretical Chemistry
  • Organic Chemistry
  • Inorganic Chemistry

Cite this

Expression and Activity of TRPA1 and TRPV1 in the Intervertebral Disc : Association with Inflammation and Matrix Remodeling. / Kameda, Takuya; Zvick, Joel; Vuk, Miriam; Sadowska, Aleksandra; Tam, Wai Kit; Leung, Victor Y.; Bölcskei, K.; Helyes, Z.; Applegate, Lee Ann; Hausmann, Oliver N.; Klasen, Juergen; Krupkova, Olga; Wuertz-Kozak, Karin.

In: International journal of molecular sciences, Vol. 20, No. 7, 10.04.2019.

Research output: Contribution to journalArticle

Kameda, T, Zvick, J, Vuk, M, Sadowska, A, Tam, WK, Leung, VY, Bölcskei, K, Helyes, Z, Applegate, LA, Hausmann, ON, Klasen, J, Krupkova, O & Wuertz-Kozak, K 2019, 'Expression and Activity of TRPA1 and TRPV1 in the Intervertebral Disc: Association with Inflammation and Matrix Remodeling', International journal of molecular sciences, vol. 20, no. 7. https://doi.org/10.3390/ijms20071767
Kameda, Takuya ; Zvick, Joel ; Vuk, Miriam ; Sadowska, Aleksandra ; Tam, Wai Kit ; Leung, Victor Y. ; Bölcskei, K. ; Helyes, Z. ; Applegate, Lee Ann ; Hausmann, Oliver N. ; Klasen, Juergen ; Krupkova, Olga ; Wuertz-Kozak, Karin. / Expression and Activity of TRPA1 and TRPV1 in the Intervertebral Disc : Association with Inflammation and Matrix Remodeling. In: International journal of molecular sciences. 2019 ; Vol. 20, No. 7.
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AU - Vuk, Miriam

AU - Sadowska, Aleksandra

AU - Tam, Wai Kit

AU - Leung, Victor Y.

AU - Bölcskei, K.

AU - Helyes, Z.

AU - Applegate, Lee Ann

AU - Hausmann, Oliver N.

AU - Klasen, Juergen

AU - Krupkova, Olga

AU - Wuertz-Kozak, Karin

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N2 - Transient receptor potential (TRP) channels have emerged as potential sensors and transducers of inflammatory pain. The aims of this study were to investigate (1) the expression of TRP channels in intervertebral disc (IVD) cells in normal and inflammatory conditions and (2) the function of Transient receptor potential ankyrin 1 (TRPA1) and Transient receptor potential vanilloid 1 (TRPV1) in IVD inflammation and matrix homeostasis. RT-qPCR was used to analyze human fetal, healthy, and degenerated IVD tissues for the gene expression of TRPA1 and TRPV1. The primary IVD cell cultures were stimulated with either interleukin-1 beta (IL-1β) or tumor necrosis factor alpha (TNF-α) alone or in combination with TRPA1/V1 agonist allyl isothiocyanate (AITC, 3 and 10 µM), followed by analysis of calcium flux and the expression of inflammation mediators (RT-qPCR/ELISA) and matrix constituents (RT-qPCR). The matrix structure and composition in caudal motion segments from TRPA1 and TRPV1 wild-type (WT) and knock-out (KO) mice was visualized by FAST staining. Gene expression of other TRP channels (A1, C1, C3, C6, V1, V2, V4, V6, M2, M7, M8) was also tested in cytokine-treated cells. TRPA1 was expressed in fetal IVD cells, 20% of degenerated IVDs, but not in healthy mature IVDs. TRPA1 expression was not detectable in untreated cells and it increased upon cytokine treatment, while TRPV1 was expressed and concomitantly reduced. In inflamed IVD cells, 10 µM AITC activated calcium flux, induced gene expression of IL-8, and reduced disintegrin and metalloproteinase with thrombospondin motifs 5 (ADAMTS5) and collagen 1A1, possibly via upregulated TRPA1. TRPA1 KO in mice was associated with signs of degeneration in the nucleus pulposus and the vertebral growth plate, whereas TRPV1 KO did not show profound changes. Cytokine treatment also affected the gene expression of TRPV2 (increase), TRPV4 (increase), and TRPC6 (decrease). TRPA1 might be expressed in developing IVD, downregulated during its maturation, and upregulated again in degenerative disc disease, participating in matrix homeostasis. However, follow-up studies with larger sample sizes are needed to fully elucidate the role of TRPA1 and other TRP channels in degenerative disc disease.

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KW - collagen

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KW - TRPC6

KW - TRPV1

KW - TRPV2

KW - TRPV4

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