Tm4sf19 inhibition ameliorates inflammation and bone destruction in collagen-induced arthritis by suppressing TLR4-mediated inflammatory signaling and abnormal osteoclast activation

Abstract

Rheumatoid arthritis (RA) is an autoimmune disease characterized by inflammation and abnormal osteoclast activation, leading to bone destruction. We previously demonstrated that the large extracellular loop (LEL) of Tm4sf19 is important for its function in osteoclast differentiation, and LEL-Fc, a competitive inhibitor of Tm4sf19, effectively suppresses osteoclast multinucleation and prevent bone loss associated with osteoporosis. This study aimed to investigate the role of Tm4sf19 in RA, an inflammatory and abnormal osteoclast disease, using a mouse model of collagen-induced arthritis (CIA). Tm4sf19 expression was observed in macrophages and osteoclasts within the inflamed synovium, and Tm4sf19 expression was increased together with inflammatory genes in the joint bones of CIA-induced mice compared with the sham control group. Inhibition of Tm4sf19 by LEL-Fc demonstrated both preventive and therapeutic effects in a CIA mouse model, reducing the CIA score, swelling, inflammation, cartilage damage, and bone damage. Knockout of Tm4sf19 gene or inhibition of Tm4sf19 activity by LEL-Fc suppressed LPS/IFN-γ-induced TLR4-mediated inflammatory signaling in macrophages. LEL-Fc disrupted not only the interaction between Tm4sf19 and TLR4/MD2, but also the interaction between TLR4 and MD2. μCT analysis showed that LEL-Fc treatment significantly reduced joint bone destruction and bone loss caused by hyperactivated osteoclasts in CIA mice. Taken together, these findings suggest that LEL-Fc may be a potential treatment for RA and RA-induced osteoporosis by simultaneously targeting joint inflammation and bone destruction caused by abnormal osteoclast activation.

Keywords

Medical and Health Sciences / Clinical Sciences

References

[1] LlorenteI, Garcia-CastanedaN, ValeroC, Gonzalez-AlvaroI, CastanedaS. Osteoporosis in rheumatoid arthritis: dangerous liaisons. Front Med., 2020, 7. 601618

[2] MeirowY, et al. . Specific inflammatory osteoclast precursors induced during chronic inflammation give rise to highly active osteoclasts associated with inflammatory bone loss. Bone Res., 2022, 10: 36.

[3] NiuQ, GaoJ, WangL, LiuJ, ZhangL. Regulation of differentiation and generation of osteoclasts in rheumatoid arthritis. Front Immunol., 2022, 13. 1034050

[4] KomatsuN, TakayanagiH. Mechanisms of joint destruction in rheumatoid arthritis - immune cell-fibroblast-bone interactions. Nat. Rev. Rheumatol., 2022, 18: 415-429.

[5] LewisMJ, et al. . Molecular portraits of early rheumatoid arthritis identify clinical and treatment response phenotypes. Cell Rep., 2019, 28: 2455-2470.e2455.

[6] BiH, et al. . Key triggers of osteoclast-related diseases and available strategies for targeted therapies: a review. Front Med. (Lausanne), 2017, 4: 234.

[7] DanksL, et al. . RANKL expressed on synovial fibroblasts is primarily responsible for bone erosions during joint inflammation. Ann. Rheum. Dis., 2016, 75: 1187-1195.

[8] Kloc, M., Subuddhi, A., Uosef, A., Kubiak, J. Z. & Ghobrial, R. M. Monocyte-macrophage lineage cell fusion. Int. J. Mol. Sci.23, 6553 (2022).

[9] LevaotN, et al. . Osteoclast fusion is initiated by a small subset of RANKL-stimulated monocyte progenitors, which can fuse to RANKL-unstimulated progenitors. Bone, 2015, 79: 21-28.

[10] Blangy, A. et al. The osteoclast cytoskeleton - current understanding and therapeutic perspectives for osteoporosis. J. Cell. Sci. 133, jcs244798 (2020).

[11] HuangQQ, PopeRM. The role of toll-like receptors in rheumatoid arthritis. Curr. Rheumatol. Rep., 2009, 11: 357-364.

[12] Satoh, T. & Akira, S. Toll-like receptor signaling and its inducible proteins. Microbiol. Spectr.4, https://doi.org/10.1128/microbiolspec.MCHD-0040-2016 (2016).

[13] SouzaPPC, LernerUH. Finding a toll on the route: the fate of osteoclast progenitors after toll-like receptor activation. Front Immunol., 2019, 10: 1663.

[14] RoelofsMF, et al. . Type I interferons might form the link between Toll-like receptor (TLR) 3/7 and TLR4-mediated synovial inflammation in rheumatoid arthritis (RA). Ann. Rheum. Dis., 2009, 68: 1486-1493.

[15] ZhuX, et al. . The critical role of toll-like receptor 4 in bone remodeling of osteoporosis: from inflammation recognition to immunity. Front Immunol., 2024, 15. 1333086

[16] BartelsYL, et al. . Inhibition of TLR4 signalling to dampen joint inflammation in osteoarthritis. Rheumatology, 2024, 63: 608-618.

[17] Abdollahi-RoodsazS, et al. . Inhibition of Toll-like receptor 4 breaks the inflammatory loop in autoimmune destructive arthritis. Arthritis Rheum., 2007, 56: 2957-2967.

[18] van DeventerS, ArpAB, van SprielAB. Dynamic plasma membrane organization: a complex symphony. Trends Cell Biol., 2021, 31: 119-129.

[19] KovalenkoOV, MetcalfDG, DeGradoWF, HemlerME. Structural organization and interactions of transmembrane domains in tetraspanin proteins. BMC Struct. Biol., 2005, 5. 11

[20] DingL, et al. . TM4SF19 aggravates LPS-induced attenuation of vascular endothelial cell adherens junctions by suppressing VE-cadherin expression. Biochem Biophys Res. Commun., 2020, 533: 1204-1211.

[21] ChoiC, et al. . TM4SF19-mediated control of lysosomal activity in macrophages contributes to obesity-induced inflammation and metabolic dysfunction. Nat. Commun., 2024, 15. 2779

[22] ParkS, et al. . Tm4sf19 deficiency inhibits osteoclast multinucleation and prevents bone loss. Metabolism, 2024, 151. 155746

[23] GuoX, et al. . Pharmacodynamic biomarkers and differential effects of TNF- and GM-CSF-targeting biologics in rheumatoid arthritis. Int. J. Rheum. Dis., 2019, 22: 646-653.

[24] ChengL, et al. . New insights from single-cell sequencing data: synovial fibroblasts and synovial macrophages in rheumatoid arthritis. Front Immunol., 2021, 12. 709178

[25] SaekiN, ImaiY. Reprogramming of synovial macrophage metabolism by synovial fibroblasts under inflammatory conditions. Cell Commun. Signal, 2020, 18. 188

[26] HasegawaT, et al. . Identification of a novel arthritis-associated osteoclast precursor macrophage regulated by FoxM1. Nat. Immunol., 2019, 20: 1631-1643.

[27] Li, J. et al. BAD inactivation exacerbates rheumatoid arthritis pathology by promoting survival of sublining macrophages. Elife9, e56309 (2020).

[28] ParkBS, et al. . The structural basis of lipopolysaccharide recognition by the TLR4-MD-2 complex. Nature, 2009, 458: 1191-1195.

[29] CutoloM, CampitielloR, GotelliE, SoldanoS. The Role of M1/M2 macrophage polarization in rheumatoid arthritis synovitis. Front Immunol., 2022, 13. 867260

[30] PerilliE, et al. . Quantifying not only bone loss, but also soft tissue swelling, in a murine inflammatory arthritis model using micro-computed tomography. Scand J. Immunol., 2015, 81: 142-150.

[31] HumphreyMB, NakamuraMC. A comprehensive review of immunoreceptor regulation of osteoclasts. Clin. Rev. Allergy. Immunol., 2016, 51: 48-58.

[32] BrooksPJ, GlogauerM, McCullochCA. An overview of the derivation and function of multinucleated giant cells and their role in pathologic processes. Am. J. Pathol., 2019, 189: 1145-1158.

[33] WangX, YamauchiK, MitsunagaT. A review on osteoclast diseases and osteoclastogenesis inhibitors recently developed from natural resources. Fitoterapia, 2020, 142. 104482

[34] Suzuki, A., Minamide, M., Iwaya, C., Ogata, K. & Iwata, J. Role of metabolism in bone development and homeostasis. Int. J. Mol. Sci.21, 8992 (2020).

[35] SunY, et al. . Macrophage-osteoclast associations: origin, polarization, and subgroups. Front Immunol., 2021, 12. 778078

[36] AhmadzadehK, VanoppenM, RoseCD, MatthysP, WoutersCH. Multinucleated giant cells: current insights in phenotype, biological activities, and mechanism of formation. Front Cell Dev. Biol., 2022, 10. 873226

[37] SamarpitaS, KimJY, RasoolMK, KimKS. Investigation of toll-like receptor (TLR) 4 inhibitor TAK-242 as a new potential anti-rheumatoid arthritis drug. Arthritis Res. Ther., 2020, 22: 16.

[38] HouL, SasakiH, StashenkoP. Toll-like receptor 4-deficient mice have reduced bone destruction following mixed anaerobic infection. Infect Immun., 2000, 68: 4681-4687.

[39] NakamuraH, et al. . Lack of Toll-like receptor 4 decreases lipopolysaccharide-induced bone resorption in C3H/HeJ mice in vivo. Oral Microbiol. Immunol., 2008, 23: 190-195.

[40] CaoY, et al. . TLR4 knockout ameliorates streptozotocin-induced osteoporosis in a mouse model of diabetes. Biochem. Biophys. Res. Commun., 2021, 546: 185-191.

[41] VijayanV, KhandelwalM, ManglaniK, GuptaS, SuroliaA. Methionine down-regulates TLR4/MyD88/NF-kappaB signalling in osteoclast precursors to reduce bone loss during osteoporosis. Br. J. Pharmacol., 2014, 171: 107-121.

[42] ZhangY, ZhaoQ. AEG-1 deletion promotes cartilage repair and modulates bone remodeling-related cytokines via TLR4/MyD88/NF-kappaB inhibition in ovariectomized rats with osteoporosis. Ann. Transl. Med., 2020, 8: 1298.

[43] XieH, CaoL, YeL, ShanG, SongW. The miR-1906 mimic attenuates bone loss in osteoporosis by down-regulating the TLR4/MyD88/NF-kappaB pathway. Physiol. Int., 2021, 107: 469-478.

[44] LuQ, et al. . Patchouli alcohol modulates the pregnancy X Receptor/Toll-like Receptor 4/Nuclear Factor Kappa B axis to suppress osteoclastogenesis. Front Pharmacol., 2021, 12. 684976

[45] Morshed, A., Abbas, A. B., Hu, J. & Xu, H. Shedding new light on the role of alphanubeta3 and alpha5beta1 Integrins in rheumatoid arthritis. Molecules24, 1537 (2019).

[46] YangD, WanY. Molecular determinants for the polarization of macrophage and osteoclast. Semin Immunopathol, 2019, 41: 551-563.

[47] HasegawaT, IshiiM. Pathological osteoclasts and precursor macrophages in inflammatory arthritis. Front Immunol., 2022, 13. 867368

[48] PerisP, MonegalA, GuanabensN. Bisphosphonates in inflammatory rheumatic diseases. Bone, 2021, 146. 115887

[49] LauperK, et al. . Effectiveness of TNF-inhibitors, abatacept, IL6-inhibitors and JAK-inhibitors in 31 846 patients with rheumatoid arthritis in 19 registers from the ‘JAK-pot’ collaboration. Ann. Rheum. Dis., 2022, 81: 1358-1366.

[50] HuQ, ZhongX, TianH, LiaoP. The efficacy of denosumab in patients with rheumatoid arthritis: a systematic review and pooled analysis of randomized or matched data. Front Immunol., 2021, 12. 799575

[51] ReddyV, CohenS. Role of Janus Kinase inhibitors in rheumatoid arthritis treatment. Curr. Opin. Rheumatol., 2021, 33: 300-306.

[52] BreuilV, Euller-ZieglerL. Bisphosphonate therapy in rheumatoid arthritis. Joint Bone Spine, 2006, 73: 349-354.

Funding

Korea Drug Development Fund (KDDF)(RS-2023-00282595)

RIGHTS & PERMISSIONS

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