Antisense Oligonucleotide Blockade of Tumor Necrosis Factor-α in Two Murine Models of Colitis

Abstract

Tumor necrosis factor-␣ (TNF-␣) is a key cytokine involved in the pathogenesis of inflammatory bowel disease. We have developed a second-generation antisense oligonucleotide (ISIS 25302) specific for murine TNF-␣ and have evaluated this oligonucleotide in two models of gut inflammation of distinct etiology. ISIS 25302 decreased TNF-␣ mRNA in a dose-and sequence-dependent manner in vitro in the mouse macrophage cell line P388D1. It also reduced TNF-␣ mRNA in vivo, in whole adipose tissue and in macrophages isolated from the adipose tissue of db/db mice, a strain with constitutively high expression of TNF-␣. ISIS 25302 significantly reduced disease activity index scores in mice with both an acute and a chronic form of dextran sodium sulfate (DSS)-induced colitis. It also significantly improved histopathological scores in interleukin (IL)-10-deficient mice. This was accompanied by reductions in both the basal and lipopolysaccharide-stimulated secretion of TNF-␣ and interferon-␥ in colonic organ cultures from IL-10 Ϫ/Ϫ mice. In this model, efficacy was obtained with both a prophylactic treatment regimen or a therapeutic dosing protocol begun after colitis was already present. In both the DSS and IL-10 Ϫ/Ϫ models, scrambled and mismatch control oligonucleotides were largely without effect, suggesting that ISIS 25302 was exerting its effects through a sequence-dependent antisense mechanism.

Absence of bacterially induced RELMβ reduces injury in the dextran sodium sulfate model of colitis

“…Thus, recombinant RELMβ activated the production of the proinflammatory cytokine TNF-α by macrophages and induced inflammatory cell infiltration when injected locally. The ability of RELMβ to activate the expression of TNF-α, an upstream mediator of the general inflammatory response, is consistent with the previously defined role of macrophages and TNF-α in DSS-induced colitis (20,22). This ability also correlates well with the general decrease in the inflammatory infiltrate observed in RELMβ -/-mice treated with DSS for 7 days compared with similarly treated wild-type mice ( Figure 4 and Table 1).…”

“…Importantly, survival after DSS administration was dramatically enhanced in RELMβ -/-mice ( Figure 2C). Consistent with the evidence for reduced injury, the percentage of the colonic mucosa containing ulcerations (Figure 2D) and the mRNA expression of an important mediator of inflammation in both patients with IBD and mice with DSSinduced colitis, TNF-α (19,20), were significantly reduced in RELMβ -/-mice ( Figure 2E). Taken together, these results provide compelling clinical evidence that RELMβ -/-mice are significantly less susceptible to DSS-induced colitis.…”

“…To determine whether RELMβ could activate macrophages in vitro, a multiplex assessment of protein secretion induced by the treatment of thioglycolate-isolated macrophages with recombinant RELMβ revealed the specific induction of only 2 proteins, TNF-α and IL-15 ( Figure 6A), out of 22 cytokines and chemokines assayed (IL-1α, IL-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12p70, IL-13, IL-17, G-CSF, GM-CSF, IFN-γ, IP-10, KC, MCP-1, MIP-1α, and RANTES). Interestingly, both TNF-α and IL-15 have been previously shown to play an important role in DSS-induced colitis (20,24). Although TNF-α levels were reduced in RELMβ -/-compared with wild-type mice treated with DSS ( Figure 2E), there was no difference in the mRNA expression of IL-15 (data not shown).…”

RNA interference-based nanosystems for inflammatory bowel disease therapy

“…In subsequent in vivo experiments, the administration of anti-TNF-α oligonucleotide via injection resulted in reduced disease activity index scores in both acute and chronic murine models of colitis. 28 Furthermore, TNF-α siRNA liposomes (Lipofectamine 2000; Invitrogen) were rectally instilled in a murine model of IBD. The experimental results showed a relative mucosal resistance to experimental colitis.…”

Effects of the Sijunzi decoction on the immunological function in rats with dextran sulfate-induced ulcerative colitis

“…The distal 8 cm of the colon was excised, opened by longitudinal incision and rinsed with saline. Sections of the colon were fixed in 4% polyformaldehyde for histopathological evaluation of the CMDI score (Table II) (10). The remaining samples were stored at -80˚C for subsequent analyses.…”