The role of TNFRSF11B in development of osteoarthritic cartilage

Abstract Objectives OA is a complex genetic disease with different risk factors contributing to its development. One of the genes, TNFRSF11B, previously identified with gain-of-function mutation in a family with early-onset OA with chondrocalcinosis, is among the highest upregulated genes in lesioned OA cartilage (RAAK-study). Here, we determined the role of TNFRSF11B overexpression in development of OA. Methods Human primary articular chondrocytes (9 donors RAAK study) were transduced using lentiviral particles with or without TNFRSF11B. Cells were cultured for 1 week in a 3 D in-vitro chondrogenic model. TNFRSF11B overexpression was confirmed by RT-qPCR, immunohistochemistry and ELISA. Effects of TNFRSF11B overexpression on cartilage matrix deposition, matrix mineralization, and genes highly correlated to TNFRSF11B in RNA-sequencing dataset (r >0.75) were determined by RT-qPCR. Additionally, glycosaminoglycans and collagen deposition were visualized with Alcian blue staining and immunohistochemistry (COL1 and COL2). Results Overexpression of TNFRSF11B resulted in strong upregulation of MMP13, COL2A1 and COL1A1. Likewise, mineralization and osteoblast characteristic markers RUNX2, ASPN and OGN showed a consistent increase. Among 30 genes highly correlated to TNFRSF11B, expression of only eight changed significantly, with BMP6 showing the highest increase (9-fold) while expression of RANK and RANKL remained unchanged indicating previously unknown downstream pathways of TNFRSF11B in cartilage. Conclusion Results of our 3D in vitro chondrogenesis model indicate that upregulation of TNFRSF11B in lesioned OA cartilage may act as a direct driving factor for chondrocyte to osteoblast transition observed in OA pathophysiology. This transition does not appear to act via the OPG/RANK/RANKL triad common in bone remodeling.

[1]  H. Komori,et al.  Runx2 is essential for the transdifferentiation of chondrocytes into osteoblasts , 2020, PLoS genetics.

[2]  Yunliang Guo,et al.  CCAL1 enhances osteoarthritis through the NF‐κB/AMPK signaling pathway , 2020, FEBS open bio.

[3]  Gyeong Min Kim,et al.  Osteoclast-associated receptor blockade prevents articular cartilage destruction via chondrocyte apoptosis regulation , 2020, Nature Communications.

[4]  Charles A. Harris,et al.  Adipose tissue is a critical regulator of osteoarthritis , 2020, Proceedings of the National Academy of Sciences.

[5]  M. Reinders,et al.  RNA sequencing data integration reveals an miRNA interactome of osteoarthritis cartilage , 2018, Annals of the rheumatic diseases.

[6]  Damian Szklarczyk,et al.  STRING v11: protein–protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets , 2018, Nucleic Acids Res..

[7]  Huidi Jiang,et al.  Regulation and biological role of the peptide/histidine transporter SLC15A3 in Toll-like receptor-mediated inflammatory responses in macrophage , 2018, Cell Death & Disease.

[8]  J. B. S. Garcia,et al.  Strontium ranelate as a possible disease-modifying osteoarthritis drug: a systematic review , 2018, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[9]  S. Mohan,et al.  The art of building bone: emerging role of chondrocyte-to-osteoblast transdifferentiation in endochondral ossification , 2018, Bone Research.

[10]  E. Mitton-Fitzgerald,et al.  Mutations in osteoprotegerin account for the CCAL1 locus in calcium pyrophosphate deposition disease. , 2018, Osteoarthritis and cartilage.

[11]  C. Rosen,et al.  Intracellular lipid droplets support osteoblast function , 2017, Adipocyte.

[12]  C. P. Winlove,et al.  Lipid distribution, composition and uptake in bovine articular cartilage studied using Raman micro‐spectrometry and confocal microscopy , 2017, Journal of anatomy.

[13]  F. Tian,et al.  Strontium ranelate causes osteophytes overgrowth in a model of early phase osteoarthritis , 2017, BMC Musculoskeletal Disorders.

[14]  Di Chen,et al.  Osteoarthritis: toward a comprehensive understanding of pathological mechanism , 2017, Bone Research.

[15]  J. Oliveira,et al.  Basic science of osteoarthritis , 2016, Journal of Experimental Orthopaedics.

[16]  J. Schwartz,et al.  Gene expression changes in damaged osteoarthritic cartilage identify a signature of non-chondrogenic and mechanical responses , 2016, Osteoarthritis and cartilage.

[17]  Jerry C. Hu,et al.  Initiation of Chondrocyte Self-Assembly Requires an Intact Cytoskeletal Network. , 2016, Tissue engineering. Part A.

[18]  P. Slagboom,et al.  A gain of function mutation in TNFRSF11B encoding osteoprotegerin causes osteoarthritis with chondrocalcinosis , 2014, Annals of the rheumatic diseases.

[19]  F. Verbeek,et al.  Underlying molecular mechanisms of DIO2 susceptibility in symptomatic osteoarthritis , 2014, Annals of the rheumatic diseases.

[20]  M. Brandi,et al.  A review on strontium ranelate long-term antifracture efficacy in the treatment of postmenopausal osteoporosis , 2013, Therapeutic advances in musculoskeletal disease.

[21]  F. Lafeber,et al.  Strontium ranelate: ready for clinical use as disease-modifying osteoarthritis drug? , 2022 .

[22]  M. Coolsen,et al.  Redifferentiation of dedifferentiated human articular chondrocytes: comparison of 2D and 3D cultures. , 2012, Osteoarthritis and cartilage.

[23]  M. Brown,et al.  Novel ANKH Amino Terminus Mutation (Pro5Ser) Associated With Early-Onset Calcium Pyrophosphate Disease With Associated Phosphaturia , 2012, Journal of clinical rheumatology : practical reports on rheumatic & musculoskeletal diseases.

[24]  M. Doherty,et al.  Pathophysiology of articular chondrocalcinosis—role of ANKH , 2011, Nature Reviews Rheumatology.

[25]  J. Pelletier,et al.  New perspective in osteoarthritis: the OPG and RANKL system as a potential therapeutic target? , 2009, The Keio journal of medicine.

[26]  T. Muneta,et al.  Prevention of cartilage destruction with intraarticular osteoclastogenesis inhibitory factor/osteoprotegerin in a murine model of osteoarthritis. , 2007, Arthritis and rheumatism.

[27]  S. Théoleyre,et al.  The molecular triad OPG/RANK/RANKL: involvement in the orchestration of pathophysiological bone remodeling. , 2004, Cytokine & growth factor reviews.

[28]  J. Pelletier,et al.  The inhibition of subchondral bone resorption in the early phase of experimental dog osteoarthritis by licofelone is associated with a reduction in the synthesis of MMP-13 and cathepsin K. , 2004, Bone.

[29]  M. Lotz,et al.  The osteoprotegerin/receptor activator of nuclear factor kappaB/receptor activator of nuclear factor kappaB ligand system in cartilage. , 2001, Arthritis and rheumatism.

[30]  S. Takeshita,et al.  The Transition of Cadherin Expression in Osteoblast Differentiation from Mesenchymal Cells: Consistent Expression of Cadherin‐11 in Osteoblast Lineage , 2001, Journal of bone and mineral research : the official journal of the American Society for Bone and Mineral Research.