A Matrix Metalloproteinase 1–Cleavable Composite Peptide Derived From Transforming Growth Factor β–Inducible Gene h3 Potently Inhibits Collagen‐Induced Arthritis

Abstract

Objective. Transforming growth factor ␤-inducible gene h3 (␤IG-H3), which is abundantly expressed in rheumatoid synovium, and matrix metalloproteinases (MMP) play important roles in the pathogenesis of rheumatoid arthritis (RA). The aim of this study was to determine the therapeutic efficacy of ␤IG-H3-derived peptides using MMP-1-dependent target tissue delivery in chronic inflammatory arthritis.Methods. Peptides developed from ␤IG-H3 derivatives, including the second and fourth YH peptides, the fourth fas-1 domain, the fourth fas-1 domain truncated for H1 and H2 sequences (dhfas-1), and an MMP-1-cleavable composite peptide (MFK24), were cloned. We confirmed the specificity of MFK24 cleavage by immunoblot analysis after treatment with different proteases.Results. The YH18 peptide in the fourth fas-1 domain of ␤IG-H3 was weakly effective in suppressing arthritis severity in mice with collagen-induced arthritis (CIA). Treatment with higher-dose dhfas-1 (30 mg/kg) showed remarkable efficacy, whereas treatment with a lower dose (10 mg/kg) resulted in only partial improvement. MFK24, a composite peptide consisting of dhfas-1 and RGD peptide linked by MMP-1 substrate, was cleaved specifically by MMP-1. The adhesion and migration of NIH3T3 cells mediated by ␤IG-H3 were inhibited by MFK24 at a low concentration. MFK24 suppressed the adhesion of NIH3T3 cells more efficiently compared with murine dhfas-1 (MFK00) or RGD, either alone or in combination. The therapeutic efficacy of MFK24 in mice with CIA was remarkably enhanced, with consistently reduced expression of inflammatory mediators within joint tissue.Conclusion. This proof-of-concept study showed that an MMP-cleavable composite peptide, based on ␤IG-H3 derivatives, had markedly improved therapeutic efficacy in chronic inflammatory arthritis, implicating a new expandable strategy for enhancement of the efficacy of 2 different active molecules in RA.Rheumatoid arthritis (RA) is a chronic destructive arthritis characterized by proliferating pannus, in which synoviocyte proliferation, increased expression of extracellular matrix (ECM) proteins, immune cell infiltration, and neovascularization are characteristic findings (1-3). Fibroblast-like synoviocytes (FLS) serve as one of the primary players in joint destruction by producing proteolytic enzymes, including matrix metalloproteinases (MMPs) (4), via interactions with abundantly expressed ECM proteins (5-7) and inflammatory cytokines (8).Transforming growth factor ␤-inducible gene h3 (␤IG-H3) is an ECM protein that is secreted by several cell types, such as FLS, in synovial tissue and is highly expressed in rheumatoid synovium (9,10). The ␤IG-H3 protein, which is composed of 4 homologous fas-1 domains and an Arg-Gly-Asp (RGD) motif at the carboxyl terminal,

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“…4,26,41 Therefore, identification of a comparable effect for DEX and a low dose of DEX-PEG-coated CNT on the expression of MMP-1 and MMP-3 is essential. Real-time PCR showed that both DEX and DEX-PEG-coated CNT diminished TNF-α-induced expression of MMP-1 and MMP-3.…”

Tumor necrosis factor receptor-associated factor 6 promotes migration of rheumatoid arthritis fibroblast-like synoviocytes

“…Sixteen 10-week-old male DBA/1 mice were purchased from the Chinese Academy of Sciences (Shanghai, China) and were maintained under a controlled temperature (22˚C) and a 12-h light/dark cycle (light, 0700-1900 h), with access to a water and a standard diet ad libitum. Mice were immunized with Freund's adjuvant (Sigma-Aldrich, St. Louis, MO, USA)) and 100 µg bovine type II collagen (Chondrex, Redmond, WA, USA) at the base of the tail on day 0 and day 21 as previously described (7,8). Animal experimental protocols were approved by the Animal Committee of Zaozhuang Municipal Hospital (Zaozhuang, China).…”