Protection against collagen-induced arthritis by intramuscular gene therapy with an expression plasmid for the interleukin-1 receptor antagonist

Abstract

The interleukin-1 receptor antagonist (IL-1Ra) is an endogenous protein that can prevent the binding of IL-1 to its cellsurface receptors. Among a number of techniques for gene transfer in vivo, the direct injection of naked DNA into muscle is simple, inexpensive and safe. In this study, we evaluated the potential of intramuscular gene therapy with plasmid DNA containing the cDNA for IL-1Ra in the prevention of murine collagen-induced arthritis (CIA). DBA/1 mice were immunized with bovine type II collagen. At 4 weeks after the initial immunization, expression plasmid for IL-1Ra was injected into four selected sites in the thigh and calf muscles of DBA/1 mice. Control mice received the same plasmid, but lacking the IL-1Ra coding sequence. Macroscopic analysis of paws for redness, swelling and deformities showed that the onset of moderate to severe CIA in the paws of mice injected with IL-1Ra DNA was significantly prevented (Po0.05). In addition, both the synovitis and the cartilage erosion in knee joints were dramatically reduced in mice treated with IL-1Ra DNA (Po0.05). The expression of IL-1b was significantly decreased in the ankle joints of mice treated with IL-1Ra (Po0.01). Interestingly, the levels of IL-1Ra in sera and joints after intramuscular injection of IL-1Ra DNA were significantly lower than when protein had been used in previous reports, suggesting that the therapeutic effect may be achieved by an alternative mechanism(s) rather than by systemic elevation of IL-1Ra. These observations provide the first evidence that direct intramuscular injection of expression plasmid for IL-1Ra may effectively suppress the inflammatory pathology in arthritis.

Naked DNA expressing two isoforms of hepatocyte growth factor induces collateral artery augmentation in a rabbit model of limb ischemia

“…One possible explanation for this decrease is the gradual degradation of exogenously added DNA as time progresses. 38 The HGF level in the serum and other organs after DNA injection was very low or undetectable even when a high level of HGF protein was readily detected in the injected muscle. This is somewhat different from the previous observation that a significant level of serum interleukin-1 receptor antagonist (IL-1Ra) protein could be found after intramuscular administration of IL-1Ra DNA.…”

“…This is somewhat different from the previous observation that a significant level of serum interleukin-1 receptor antagonist (IL-1Ra) protein could be found after intramuscular administration of IL-1Ra DNA. 38 One possible explanation for this discrepancy is the high affinity of HGF to heparan sulfate proteoglycan on the cell surface and in the extracellular matrix. Indeed, the serum level of HGF protein has been reported to increase after the injection of soluble heparin in humans.…”

“…38 For other organs, brain, heart, lung, spleen, kidney, stomach, small and large intestine, ovary and uterus (or seminal vesicle, prostate, testis and penis), the lymph node and sera were collected from rats. Then, 100 mg of each tissue was lysed with 500 ml of cell lysis buffer.…”

Gene Therapy Outcomes in Experimental Models of Inflammatory Arthritis

“…Over the past 7 years or so a considerable amount of research activity has been devoted to the development of gene therapeutic strategies that have resulted in 1) neutralization of the effects of IL-1 with an IL-1 receptor antagonist (IL-1-Ra) expression plasmid (Kim et al;; 2) elevating the levels of Th2 cytokines exemplified by IL-10 (Traister & Hirsh, 2008), IL-4 (Kageyama et al, 2004) and IL-13 (Nabbe et al, 2005); 3) suppression of Th1 cytokines such as IL-18 (Smeets et al, 2003) and IL-17, the latter by modification of the indoleamine 2,3-dioxygenase pathway (Chen et al, 2011); 4) blunting of TNF--stimulated signaling (Denys et al, 2010) and interferon-activity (Adriaansen et al;; 5) up-regulation of the protein inhibitor of activated STAT1 (PIAS) activity by stimulating the capacity of small ubiquitin-like modifier E3 ligase (SUMO E3) to alter inhibitor of κB kinase (IKK ) phosphorylation (Liu & Shuai, 2008); 6) gene transfer of genetically modified chondrocytes into cartilage defects to promote cartilage regeneration (Steinert et al, 2008); and 7) promotion of adiponectin gene expression (Ebina et al, 2009). In addition, because new blood vessel formation is also intimately involved with perpetuating the chronic state of inflammation associated with RA, experimental gene therapy strategies designed to suppress the activity of pro-angiogenesis factors such as vascular endothelial growth factor (VEGF) (Afuwape et al, 2003) and Tie-2 (Chen et al, 2005) have also been earnestly pursued.…”

Eficácia terapêutica do Metotrexato na Artrite Reumatóide depende da expressão de células T reguladoras CD39+?

“…Mais especificamente a IL-1β, promove a amplificação do processo inflamatório pela indução da produção de IL-6 (Tan, 1999), RANKL e quimiocinas como MCP-1 (CCL2) e IL-8 (Goldring et al, 2002;deMarco et al, 1990), além de ativar condrócitos por vias dependentes de MAP quinases (Geng et al, 1996). Em modelos murinos de artrite induzida por colágeno do tipo 2 ou proteoglicana, o uso do antagonista do receptor de IL-1 leva a uma diminuição do infiltrado celular inflamatório, principalmente de polimorfonucleares, e confere proteção contra lesão articular (Arner et al, 1995;Kim et al, 2003;Joosten et al, 2008;Dinarello, 2000). Dados estes que confirmam o papel das citocinas nos mecanismos inflamatórios caraterísticos da AR.…”