TRIM24-RIP3 axis perturbation accelerates osteoarthritis pathogenesis

Abstract

ObjectivesRecently, necroptosis has attracted increasing attention in arthritis research; however, it remains unclear whether its regulation is involved in osteoarthritis (OA) pathogenesis. Since receptor-interacting protein kinase-3 (RIP3) plays a pivotal role in necroptosis and its dysregulation is involved in various pathological processes, we investigated the role of the RIP3 axis in OA pathogenesis.MethodsExperimental OA was induced in wild-type or Rip3 knockout mice by surgery to destabilise the medial meniscus (DMM) or the intra-articular injection of adenovirus carrying a target gene (Ad-Rip3 and Ad-Trim24 shRNA). RIP3 expression was examined in OA cartilage from human patients; Trim24, a negative regulator of RIP3, was identified by microarray and in silico analysis. Connectivity map (CMap) and in silico binding approaches were used to identify RIP3 inhibitors and to examine their direct regulation of RIP3 activation in OA pathogenesis.ResultsRIP3 expression was markedly higher in damaged cartilage from patients with OA than in undamaged cartilage. In the mouse model, adenoviral RIP3 overexpression accelerated cartilage disruption, whereas Rip3 depletion reduced DMM-induced OA pathogenesis. Additionally, TRIM24 knockdown upregulated RIP3 expression; its downregulation promoted OA pathogenesis in knee joint tissues. The CMap approach and in silico binding assay identified AZ-628 as a potent RIP3 inhibitor and demonstrated that it abolished RIP3-mediated OA pathogenesis by inhibiting RIP3 kinase activity.ConclusionsTRIM24-RIP3 axis perturbation promotes OA chronicity by activating RIP3 kinase, suggesting that the therapeutic manipulation of this pathway could provide new avenues for treating OA.

RIP1 Perturbation Induces Chondrocyte Necroptosis and Promotes Osteoarthritis Pathogenesis via Targeting BMP7

“…Necroptosis is mediated by necrosome, a supermolecular complex which contains receptor-interacting protein kinase 1 and 3 (RIP1, RIP3), and its direct substrate mixed-lineage kinase domain-like protein (MLKL), targeting the complex to appropriate downstream effectors in the necroptosisinducing process (Cho et al, 2009;He et al, 2009;Wang et al, 2014). Previous studies have discovered a possible link between necroptotic process and cartilage injury depending on oxidative stress and cytokine release in OA, and the TRIM24-RIP3 axis was proposed to promote OA chronicity by modulating the expression of catabolic factors (Riegger and Brenner, 2019;Jeon et al, 2020;Stolberg-Stolberg et al, 2020). However, the involvement of RIP1 during OA pathogenesis still lacks direct evidence.…”

Methamphetamine exposure induces neuronal programmed necrosis by activating the receptor‐interacting protein kinase 3 ‐related signalling pathway

“…RIP3 plays a role in the convergence points of multiple necrotic cell death pathways. 24,[48][49][50][51][52][53][54] Normally, the activation of death receptors can trigger programmed necrosis, but it can also be activated through non-death receptor-dependent pathways. In the death receptor-dependent pathway, the receptor is recruited through the death domain to form complex I after ligand binding.…”

Obtusifolin, an Anthraquinone Extracted from Senna obtusifolia (L.) H.S.Irwin & Barneby, Reduces Inflammation in a Mouse Osteoarthritis Model

“…The lactate dehydrogenase (LDH) assay is commonly used to measure cell toxicity [ 44 ]. Primary chondrocytes were treated with obtusifolin (0, 25, 50, 100, and 200 μM) for 24 h. The supernatant was then collected and analyzed using an LDH-cytotoxicity assay kit (K311-400; BioVision, Inc., Milpitas CA, USA).…”

“…They were then embedded in paraffin and cut into 5 μm-thick sections. The sectioned joint samples were stained with Safranin O using a standard protocol [ 44 , 46 ].…”