TRPC6 in simulated microgravity of intervertebral disc cells

Franco‐Obregón, Alfredo; Cambria, Elena; Greutert, Helen; Wernas, Timon; Hitzl, Wolfgang; Egli, Marcel; Sekiguchi, Miho; Boos, Norbert; Hausmann, Oliver; Ferguson, Stephen J.; Kobayashi, Hiroshi; Wuertz‐Kozak, Karin

doi:10.1007/s00586-018-5688-8

Cited by 12 publications

(18 citation statements)

References 57 publications

(82 reference statements)

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“…Concerning other TRP channels that were regulated by an inflammatory environment in this study, increased TRPV4 expression/signaling in the IVD has been associated with decreased tissue osmolarity and the production of pro-inflammatory cytokines [17]. Our study provided evidence that IL-1β itself can regulate gene expression of TRPV4 in IVD cells., the Gene expression of TRPC6 was shown to be reduced in IVD cells under microgravity [18], but elevated in IVDs with increasing degeneration grade [19]. In our study, TRPC6 was downregulated by IL-1β treatment.…”

Section: Discussionsupporting

confidence: 51%

“…Interestingly, the expression and activity of certain TRP channels is altered in painful joints and IVDs [11,12]. For example, the expression of TRPV4 in human IVDs was found to be elevated in regions of aggrecan depletion [17], while the gene expression of TRPC6 was associated with the severity of disc degeneration, increased expression of IL-6, and cell senescence [18,19]. TRPA1 and TRPV1 are involved in inflammation/nociception in sensory neurons and non-neuronal tissues [20,21,22].…”

Section: Introductionmentioning

confidence: 99%

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

Kameda

Zvick

Vuk

et al. 2019

IJMS

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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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Section: Discussionsupporting

confidence: 51%

Section: Introductionmentioning

confidence: 99%

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

Kameda

Zvick

Vuk

et al. 2019

IJMS

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“…TRPC6 belongs to the TRP canonical (C) subfamily, together with five other channels (C1–6). The TRPC subfamily may be involved in mechanosensing 14,22 , as well as in pain and inflammation in joint tissues 10 . We were able to show that the mRNA expression of TRPC6 was significantly higher in degenerated IVDs than in non-degenerated IVDs and that its expression tended to increase with the IVD degeneration grade as previously reported 13 .…”

Section: Discussionmentioning

confidence: 99%

Differential regulation of TRP channel gene and protein expression by intervertebral disc degeneration and back pain

Sadowska

Hitzl

Karol

et al. 2019

Sci Rep

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Intervertebral disc (IVD) degeneration and consequent low back pain (LBP) are common and costly pathological processes that require improved treatment strategies. Transient Receptor Potential (TRP) channels constitute a family of multimodal ion channels that have recently emerged as contributors to disc pathologies and were thus proposed as potential therapeutic targets, although limited data on their presence and function in the IVD exist. The purpose of this study was to determine the mRNA and protein expression of TRP channels in non-degenerated and degenerated human IVD tissue (with different pain intensity and chronicity) using gene array, conventional qPCR and immunohistochemistry. We could demonstrate that 26 out of 28 currently known TRP channels are expressed in the IVD on the mRNA level, thereby revealing novel therapeutic candidates from the TRPC, TRPM and TRPML subfamilies. TRPC6, TRPM2 and TRPML1 displayed enhanced gene and protein expression in degenerated IVDs as compared to non-degenerated IVDs. Additionally, the gene expression of TRPC6 and TRPML1 was influenced by the IVD degeneration grade. Pain intensity and/or chronicity influenced the gene and/or protein expression of TRPC6, TRPM2 and TRML1. Interestingly, decreased gene expression of TRPM2 was observed in patients treated with steroids. This study supports the importance of TRP channels in IVD homeostasis and pathology and their possible application as pharmacological targets for the treatment of IVD degeneration and LBP. However, the exact function and activation of the highlighted TRP channels will have to be determined in future studies.

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“…Moreover, cartilage-specific TRPV4 knockout in mice reduced age-related osteoarthritis [22]. The exploration of TRP channels in the IVD is at its infancy, with only few published studies [23][24][25][26][27]. Most of the known TRP channel genes are expressed in the IVD [26].…”

Section: Introductionmentioning

confidence: 99%

TRPV4 Inhibition and CRISPR-Cas9 Knockout Reduce Inflammation Induced by Hyperphysiological Stretching in Human Annulus Fibrosus Cells

Cambria

Arlt

Wandel

et al. 2020

Cells

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Mechanical loading and inflammation interact to cause degenerative disc disease and low back pain (LBP). However, the underlying mechanosensing and mechanotransductive pathways are poorly understood. This results in untargeted pharmacological treatments that do not take the mechanical aspect of LBP into account. We investigated the role of the mechanosensitive ion channel TRPV4 in stretch-induced inflammation in human annulus fibrosus (AF) cells. The cells were cyclically stretched to 20% hyperphysiological strain. TRPV4 was either inhibited with the selective TRPV4 antagonist GSK2193874 or knocked out (KO) via CRISPR-Cas9 gene editing. The gene expression, inflammatory mediator release and MAPK pathway activation were analyzed. Hyperphysiological cyclic stretching significantly increased the IL6, IL8, and COX2 mRNA, PGE2 release, and activated p38 MAPK. The TRPV4 pharmacological inhibition significantly attenuated these effects. TRPV4 KO further prevented the stretch-induced upregulation of IL8 mRNA and reduced IL6 and IL8 release, thus supporting the inhibition data. We provide novel evidence that TRPV4 transduces hyperphysiological mechanical signals into inflammatory responses in human AF cells, possibly via p38. Additionally, we show for the first time the successful gene editing of human AF cells via CRISPR-Cas9. The pharmacological inhibition or CRISPR-based targeting of TRPV4 may constitute a potential therapeutic strategy to tackle discogenic LBP in patients with AF injury.

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TRPC6 in simulated microgravity of intervertebral disc cells

Cited by 12 publications

References 57 publications

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

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

Differential regulation of TRP channel gene and protein expression by intervertebral disc degeneration and back pain

TRPV4 Inhibition and CRISPR-Cas9 Knockout Reduce Inflammation Induced by Hyperphysiological Stretching in Human Annulus Fibrosus Cells

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