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The Death Receptor 3-TNF-like protein 1A pathway drives adverse bone pathology in inflammatory arthritis.

Bull MJ, Williams AS, Mecklenburgh Z, Calder CJ, Twohig JP, Elford C, Evans BA, Rowley TF, Slebioda TJ, Taraban VY, Al-Shamkhani A, Wang EC - J. Exp. Med. (2008)

Bottom Line: In contrast, TNF-like protein 1A (TL1A), the ligand for DR3, exacerbated disease in a dose- and DR3-dependent fashion.Treatment with antagonistic anti-TL1A mAb protected animals in a systemic model of RA disease collagen-induced arthritis.We therefore conclude that the DR3-TL1A pathway regulates joint destruction in two murine models of arthritis and represents a potential novel target for therapeutic intervention in inflammatory joint disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biochemistry and Immunology, School of Medicine, Heath Park, Cardiff CF14 4XN, Wales, UK.

ABSTRACT
Rheumatoid arthritis (RA) is a chronic inflammatory disease of synovial joints that is associated with cartilage and bone destruction. Death Receptor 3 (DR3), a tumor necrosis factor (TNF) receptor superfamily member, has recently been associated with the pathogenesis of RA. We demonstrate that absence of DR3 confers resistance to the development of adverse bone pathology in experimental antigen-induced arthritis (AIA). DR3(ko) mice exhibited a reduction in all histopathological hallmarks of AIA but, in particular, failed to develop subchondral bone erosions and were completely protected from this characteristic of AIA. In contrast, TNF-like protein 1A (TL1A), the ligand for DR3, exacerbated disease in a dose- and DR3-dependent fashion. Analysis of osteoclast number within AIA joint revealed a reduction in areas susceptible to bone erosion in DR3(ko) mice, whereas in vitro osteoclastogenesis assays showed that TL1A could directly promote osteoclastogenesis in mouse and man. Treatment with antagonistic anti-TL1A mAb protected animals in a systemic model of RA disease collagen-induced arthritis. We therefore conclude that the DR3-TL1A pathway regulates joint destruction in two murine models of arthritis and represents a potential novel target for therapeutic intervention in inflammatory joint disease.

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TL1A promotes DR3-dependent in vitro osteoclastogenesis. (A) RT-PCR of DR3 in BMM. In vitro osteoclastogenesis assays were performed as described in Materials and methods. Osteoclast numbers were estimated by counting multinucleated TRAP+ cells. (B) Effect of TL1A on proportion of osteoclasts generated in presence of RANKL and M-CSF. *, P = 0.0003. Each point represents a single ivory disc from experiments on DR3wt (open symbols) or DR3ko (filled symbols) mice. Four discs from four mice were counted for each treatment. Lines mark means of graphed points. One representative experiment of two is shown. (C–E) TRAP staining of BM cells from DR3wt mice on discs with RANK-L + M-CSF and no TL1A (C) or 10 ng/ml TL1A (D) and TL1A (E), but no RANKL and M-CSF. Bars, 50 μm. (F) Toluidine blue staining of ivory discs showing pit-forming ability of osteoclasts generated in vitro. Bar, 150 μm. (G) Effect of TL1A on osteoclastogenesis from adherent human peripheral blood mononuclear cells and proportion of osteoclasts in cultures shown with (▿) or without (▴) exogenous TL1A added. **, P = 0.0013. One representative experiment of two is shown.
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fig4: TL1A promotes DR3-dependent in vitro osteoclastogenesis. (A) RT-PCR of DR3 in BMM. In vitro osteoclastogenesis assays were performed as described in Materials and methods. Osteoclast numbers were estimated by counting multinucleated TRAP+ cells. (B) Effect of TL1A on proportion of osteoclasts generated in presence of RANKL and M-CSF. *, P = 0.0003. Each point represents a single ivory disc from experiments on DR3wt (open symbols) or DR3ko (filled symbols) mice. Four discs from four mice were counted for each treatment. Lines mark means of graphed points. One representative experiment of two is shown. (C–E) TRAP staining of BM cells from DR3wt mice on discs with RANK-L + M-CSF and no TL1A (C) or 10 ng/ml TL1A (D) and TL1A (E), but no RANKL and M-CSF. Bars, 50 μm. (F) Toluidine blue staining of ivory discs showing pit-forming ability of osteoclasts generated in vitro. Bar, 150 μm. (G) Effect of TL1A on osteoclastogenesis from adherent human peripheral blood mononuclear cells and proportion of osteoclasts in cultures shown with (▿) or without (▴) exogenous TL1A added. **, P = 0.0013. One representative experiment of two is shown.

Mentions: We therefore tested the possibility that TL1A could directly promote differentiation of osteoclasts. To achieve this, we used an in vitro system of osteoclastogenesis from adherent BM-derived cells (BMC). BM macrophages (BMM) from DR3wt mice were confirmed to express DR3 (Fig. 4 A). BMC from DR3wt and DR3ko mice did not differ in their ability to generate osteoclasts in the presence of soluble RANK-L and M-CSF as measured by the formation of multinucleated TRAP+ cells (Fig. 4 B). However, TL1A addition significantly enhanced development of osteoclasts from DR3wt but not DR3ko BMC (Fig. 4, B–D). TL1A in the absence of RANK-L and M-CSF could not generate osteoclasts (Fig. 4 E). The functional capacity of in vitro–generated osteoclasts to destroy bone was visualized by toluidine blue staining of pits in the ivory discs (Fig. 4 F). This data indicates that TL1A is not necessary for osteoclastogenesis per se, but promotes it in the presence of RANK-L and M-CSF and in a DR3-dependent fashion. In support of our murine data and highlighting the significance of these results for humans, TL1A significantly promoted osteoclastogenesis from monocytes derived from human peripheral blood (Fig. 4 G).


The Death Receptor 3-TNF-like protein 1A pathway drives adverse bone pathology in inflammatory arthritis.

Bull MJ, Williams AS, Mecklenburgh Z, Calder CJ, Twohig JP, Elford C, Evans BA, Rowley TF, Slebioda TJ, Taraban VY, Al-Shamkhani A, Wang EC - J. Exp. Med. (2008)

TL1A promotes DR3-dependent in vitro osteoclastogenesis. (A) RT-PCR of DR3 in BMM. In vitro osteoclastogenesis assays were performed as described in Materials and methods. Osteoclast numbers were estimated by counting multinucleated TRAP+ cells. (B) Effect of TL1A on proportion of osteoclasts generated in presence of RANKL and M-CSF. *, P = 0.0003. Each point represents a single ivory disc from experiments on DR3wt (open symbols) or DR3ko (filled symbols) mice. Four discs from four mice were counted for each treatment. Lines mark means of graphed points. One representative experiment of two is shown. (C–E) TRAP staining of BM cells from DR3wt mice on discs with RANK-L + M-CSF and no TL1A (C) or 10 ng/ml TL1A (D) and TL1A (E), but no RANKL and M-CSF. Bars, 50 μm. (F) Toluidine blue staining of ivory discs showing pit-forming ability of osteoclasts generated in vitro. Bar, 150 μm. (G) Effect of TL1A on osteoclastogenesis from adherent human peripheral blood mononuclear cells and proportion of osteoclasts in cultures shown with (▿) or without (▴) exogenous TL1A added. **, P = 0.0013. One representative experiment of two is shown.
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fig4: TL1A promotes DR3-dependent in vitro osteoclastogenesis. (A) RT-PCR of DR3 in BMM. In vitro osteoclastogenesis assays were performed as described in Materials and methods. Osteoclast numbers were estimated by counting multinucleated TRAP+ cells. (B) Effect of TL1A on proportion of osteoclasts generated in presence of RANKL and M-CSF. *, P = 0.0003. Each point represents a single ivory disc from experiments on DR3wt (open symbols) or DR3ko (filled symbols) mice. Four discs from four mice were counted for each treatment. Lines mark means of graphed points. One representative experiment of two is shown. (C–E) TRAP staining of BM cells from DR3wt mice on discs with RANK-L + M-CSF and no TL1A (C) or 10 ng/ml TL1A (D) and TL1A (E), but no RANKL and M-CSF. Bars, 50 μm. (F) Toluidine blue staining of ivory discs showing pit-forming ability of osteoclasts generated in vitro. Bar, 150 μm. (G) Effect of TL1A on osteoclastogenesis from adherent human peripheral blood mononuclear cells and proportion of osteoclasts in cultures shown with (▿) or without (▴) exogenous TL1A added. **, P = 0.0013. One representative experiment of two is shown.
Mentions: We therefore tested the possibility that TL1A could directly promote differentiation of osteoclasts. To achieve this, we used an in vitro system of osteoclastogenesis from adherent BM-derived cells (BMC). BM macrophages (BMM) from DR3wt mice were confirmed to express DR3 (Fig. 4 A). BMC from DR3wt and DR3ko mice did not differ in their ability to generate osteoclasts in the presence of soluble RANK-L and M-CSF as measured by the formation of multinucleated TRAP+ cells (Fig. 4 B). However, TL1A addition significantly enhanced development of osteoclasts from DR3wt but not DR3ko BMC (Fig. 4, B–D). TL1A in the absence of RANK-L and M-CSF could not generate osteoclasts (Fig. 4 E). The functional capacity of in vitro–generated osteoclasts to destroy bone was visualized by toluidine blue staining of pits in the ivory discs (Fig. 4 F). This data indicates that TL1A is not necessary for osteoclastogenesis per se, but promotes it in the presence of RANK-L and M-CSF and in a DR3-dependent fashion. In support of our murine data and highlighting the significance of these results for humans, TL1A significantly promoted osteoclastogenesis from monocytes derived from human peripheral blood (Fig. 4 G).

Bottom Line: In contrast, TNF-like protein 1A (TL1A), the ligand for DR3, exacerbated disease in a dose- and DR3-dependent fashion.Treatment with antagonistic anti-TL1A mAb protected animals in a systemic model of RA disease collagen-induced arthritis.We therefore conclude that the DR3-TL1A pathway regulates joint destruction in two murine models of arthritis and represents a potential novel target for therapeutic intervention in inflammatory joint disease.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Biochemistry and Immunology, School of Medicine, Heath Park, Cardiff CF14 4XN, Wales, UK.

ABSTRACT
Rheumatoid arthritis (RA) is a chronic inflammatory disease of synovial joints that is associated with cartilage and bone destruction. Death Receptor 3 (DR3), a tumor necrosis factor (TNF) receptor superfamily member, has recently been associated with the pathogenesis of RA. We demonstrate that absence of DR3 confers resistance to the development of adverse bone pathology in experimental antigen-induced arthritis (AIA). DR3(ko) mice exhibited a reduction in all histopathological hallmarks of AIA but, in particular, failed to develop subchondral bone erosions and were completely protected from this characteristic of AIA. In contrast, TNF-like protein 1A (TL1A), the ligand for DR3, exacerbated disease in a dose- and DR3-dependent fashion. Analysis of osteoclast number within AIA joint revealed a reduction in areas susceptible to bone erosion in DR3(ko) mice, whereas in vitro osteoclastogenesis assays showed that TL1A could directly promote osteoclastogenesis in mouse and man. Treatment with antagonistic anti-TL1A mAb protected animals in a systemic model of RA disease collagen-induced arthritis. We therefore conclude that the DR3-TL1A pathway regulates joint destruction in two murine models of arthritis and represents a potential novel target for therapeutic intervention in inflammatory joint disease.

Show MeSH
Related in: MedlinePlus