TRPC1 links calcium signaling to cellular senescence in the protection against posttraumatic osteoarthritis
Meike Sambale, Starlee Lively, Osvaldo Espin-Garcia, Pratibha Potla, Chiara Pastrello, Sarah Bödecker, Linda Wessendorf, Simon Kleimann, Peter Paruzel, Rojiar Asgarian, Alexandra Tosun, Johanna Intemann, Jessica Bertrand, Francesco Dell’Accio, Mohit Kapoor, Thomas Pap, Joanna Sherwood
Meike Sambale, Starlee Lively, Osvaldo Espin-Garcia, Pratibha Potla, Chiara Pastrello, Sarah Bödecker, Linda Wessendorf, Simon Kleimann, Peter Paruzel, Rojiar Asgarian, Alexandra Tosun, Johanna Intemann, Jessica Bertrand, Francesco Dell’Accio, Mohit Kapoor, Thomas Pap, Joanna Sherwood
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Research Article Bone biology Cell biology

TRPC1 links calcium signaling to cellular senescence in the protection against posttraumatic osteoarthritis

Abstract

Transient receptor potential channel 1 (TRPC1) is a widely expressed mechanosensitive ion channel located within the endoplasmic reticulum membrane, crucial for refilling depleted internal calcium stores during activation of calcium-dependent signaling pathways. Here, we have demonstrated that TRPC1 activity is protective within cartilage homeostasis in the prevention of cellular senescence–associated cartilage breakdown during mechanical and inflammatory challenge. We revealed that TRPC1 loss is associated with early stages of osteoarthritis (OA) and plays a nonredundant role in calcium signaling in chondrocytes. Trpc1–/– mice subjected to destabilization of the medial meniscus–induced OA developed a more severe OA phenotype than WT controls. During early OA development, Trpc1–/– mice displayed an increased chondrocyte survival rate; however, remaining cells displayed features of senescence including p16INK4a expression and decreased Sox9. RNA-Seq identified differentially expressed genes related to cell number, apoptosis, and extracellular matrix organization. Trpc1–/– chondrocytes exhibited accelerated dedifferentiation, while demonstrating an increased susceptibility to cellular senescence. Targeting the mechanism of TRPC1 activation may be a promising therapeutic strategy in OA prevention.

Authors

Meike Sambale, Starlee Lively, Osvaldo Espin-Garcia, Pratibha Potla, Chiara Pastrello, Sarah Bödecker, Linda Wessendorf, Simon Kleimann, Peter Paruzel, Rojiar Asgarian, Alexandra Tosun, Johanna Intemann, Jessica Bertrand, Francesco Dell’Accio, Mohit Kapoor, Thomas Pap, Joanna Sherwood

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Figure 7

TRPC1 is required for protection against cellular senescence driven by IL-1β and during murine OA development.

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TRPC1 is required for protection against cellular senescence driven by I...
(A) Representative images of SA–β-Gal stainings (blue) of P0 WT and Trpc1–/– chondrocytes treated with 10 ng/mL IL-1β for either 24 hours, 48 hours, or for 24 hours followed by an additional 24 or 48 hours culture in control medium (rest). (B) Quantification of number of SA–β-Gal+ cells normalized to those in control untreated samples (n = 5). (C) CyQUANT fluorescence quantification of WT and Trpc1–/– chondrocytes measured at 24 and 48 hours after IL-1β stimulation to give a ratio representing proliferation rate (n = 4). (D) Western blot of p16INK4a protein levels in WT and Trpc1–/– chondrocytes treated with 10 ng/mL IL-1β for either 24 hours or for 24 hours followed by 48 hours culture in control medium. (E) Quantification of Western blotting for p16INK4a in WT and Trpc1–/– chondrocytes treated with 10 ng/mL IL-1β for either 24 hours or for 24 hours followed by 48 hours culture in control medium (n = 3). (F) CCL-2 concentration of control and IL-1β–treated chondrocyte supernatants measured by ELISA (n = 4). (G) Representative images of cleaved caspase activity in chondrocytes treated with IL-1β for 24 hours plus 48 hours rest. (H) Quantification of cleaved caspase activity in WT and Trpc1–/– chondrocytes following IL-1β stimulation (WT, n = 4; Trpc1–/–, n = 5). (I) Immunofluorescence detection of p16INK4a in medial compartments of WT and Trpc1–/– murine knee joints 2 weeks after DMM. DAPI used as a nuclear counterstain. Scale bar: 100 μm. For isotype negative control staining, see Supplemental Figure 3I. (J) Quantification of number of p16INK4a+ cells within the medial articular cartilage normalized for the total number of DAPI+ cells (n = 5). Two-way ANOVA with multiple comparisons.