title: Antisense oligonucleotide blockade of tumor necrosis factor-alpha in two murine models of colitis.
abstract: Tumor necrosis factor-alpha (TNF-alpha) 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-alpha and have evaluated this oligonucleotide in two models of gut inflammation of distinct etiology. ISIS 25302 decreased TNF-alpha mRNA in a dose- and sequence-dependent manner in vitro in the mouse macrophage cell line P388D1. It also reduced TNF-alpha 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-alpha. 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-alpha and interferon-gamma in colonic organ cultures from IL-10 -/- 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 -/- models, scrambled and mismatch control oligonucleotides were largely without effect, suggesting that ISIS 25302 was exerting its effects through a sequence-dependent antisense mechanism.
authors.affiliation: Isis Pharmaceuticals, Carlsbad, California 92008, USA. kmyers@isisph.com
authors.forename: Kathleen J
authors.initials: KJ
authors.lastname: Myers
authors.forename: Sreekant
authors.initials: S
authors.lastname: Murthy
authors.forename: Anne
authors.initials: A
authors.lastname: Flanigan
authors.forename: Donna R
authors.initials: DR
authors.lastname: Witchell
authors.forename: Madeline
authors.initials: M
authors.lastname: Butler
authors.forename: Susan
authors.initials: S
authors.lastname: Murray
authors.forename: Andrew
authors.initials: A
authors.lastname: Siwkowski
authors.forename: Deborah
authors.initials: D
authors.lastname: Goodfellow
authors.forename: Karen
authors.initials: K
authors.lastname: Madsen
authors.forename: Brenda
authors.initials: B
authors.lastname: Baker
mesh_terms: D000208:Acute Disease; D000818:Animals; D002478:Cells, Cultured; D002908:Chronic Disease; D003092:Colitis / Q000139:chemically induced / Q000235:genetics / Q000517:prevention & control*; D003106:Colon / Q000378:metabolism / Q000473:pathology; D016264:Dextran Sulfate; D016753:Interleukin-10 / Q000172:deficiency / Q000235:genetics; D007413:Intestinal Mucosa / Q000473:pathology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000378:metabolism / Q000494:pharmacology*; D054735:Phosphorothioate Oligonucleotides; D014176:Protein Biosynthesis / Q000187:drug effects; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000096:biosynthesis / Q000633:toxicity
chemical_list: C470440:ISIS 25302; D016376:Oligonucleotides, Antisense; D054735:Phosphorothioate Oligonucleotides; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016753:Interleukin-10; D016264:Dextran Sulfate