title: Rectal Delivery of a DNAzyme That Specifically Blocks the Transcription Factor GATA3 and Reduces Colitis in Mice.
abstract: GATA3 is a transcription factor that regulates T-cell production of cytokines. We investigated the role of GATA3 in development of colitis in mice.
  
  
  
  We performed quantitative polymerase chain reaction and immunofluorescence analyses of colon tissues from patients with Crohn's disease (n = 61) or ulcerative colitis (UC, n = 74) or from patients without inflammatory bowel diseases (n = 22), to measure levels of GATA3. Colitis was induced by administration of oxazolone or 2,4,6-trinitrobenzenesulfonic acid to control mice, mice with T-cell-specific deletion of GATA3, and mice with deletion of tumor necrosis factor receptor (TNFR) 1 and TNFR2 (TNFR double knockouts); some mice were given a GATA3-specific DNAzyme (hgd40) or a control DNAzyme via intrarectal administration, or systemic injections of an antibody to TNF before or during sensitization and challenge phase of colitis induction. Colon tissues were collected and immunofluorescence and histochemical analyses were performed. Lamina propria mononuclear cells and T cells were isolated and analyzed by flow cytometry or cytokine assays. Colonic distribution of labeled DNAzyme and inflammation were monitored by in vivo imaging (endoscopy) of mice.
  
  
  
  Levels of GATA3 messenger RNA were higher in colon tissues from patients with UC, but not ileal Crohn's disease, than control tissues; levels of GATA3 correlated with levels of inflammatory cytokines (interleukin [IL] 9, IL17A, IL6, IL5, IL4, IL13, and TNF). We observed increased expression of GATA3 by lamina propria T cells from mice with colitis compared with controls. Mice with T-cell-specific deletion of GATA3 did not develop colitis and their colonic tissues did not produce inflammatory cytokines (IL6, IL9, or IL13). The DNAzyme hgd40 inhibited expression of GATA3 messenger RNA by unstimulated and stimulated T cells, and distributed throughout the inflamed colons of mice with colitis. Colon tissues from mice given hgd40 had reduced expression of GATA3 messenger RNA, compared with mice given a control DNAzyme. Mice given hgd40 did not develop colitis after administration of oxazolone or 2,4,6-trinitrobenzenesulfonic acid; lamina propria cells from these mice expressed lower levels of IL6, IL9, and IL13 than cells from mice given the control DNAzyme. Mini-endoscopic images revealed that hgd40 and anti-TNF reduced colon inflammation over 3 days; hgd40 reduced colitis in TNFR double-knockout mice.
  
  
  
  Levels of GATA3 are increased in patients with UC and correlate with production of inflammatory cytokines in mice and humans. A DNAzyme that prevents expression of GATA3 reduces colitis in mice, independently of TNF, and reduces levels of cytokines in the colon. This DNAzyme might be developed for treatment of patients with UC.
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Popp
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany.
authors.forename: Katharina
authors.initials: K
authors.lastname: Gerlach
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany.
authors.forename: Stefanie
authors.initials: S
authors.lastname: Mott
authors.affiliation: sterna biologicals GmbH Co & KG, Marburg, Germany.
authors.forename: Agnieszka
authors.initials: A
authors.lastname: Turowska
authors.affiliation: Institute of Laboratory Medicine and Pathobiochemistry, Medical Faculty, Philipps University of Marburg, Marburg, Germany.
authors.forename: Holger
authors.initials: H
authors.lastname: Garn
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany.
authors.forename: Raja
authors.initials: R
authors.lastname: Atreya
authors.affiliation: Institute of Pathology, Campus Bodensee, Friedrichshafen, Germany.
authors.forename: Hans-Anton
authors.initials: HA
authors.lastname: Lehr
authors.affiliation: Harvard Medical School, Brigham and Women's Hospital, Boston, Massachusetts.
authors.forename: I-Cheng
authors.initials: IC
authors.lastname: Ho
authors.affiliation: Institute of Laboratory Medicine and Pathobiochemistry, Medical Faculty, Philipps University of Marburg, Marburg, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Renz
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany.
authors.forename: Benno
authors.initials: B
authors.lastname: Weigmann
authors.affiliation: Department of Medicine, University of Erlangen-Nürnberg, Kussmaul Research Campus, Erlangen, Germany. Electronic address: markus.neurath@uk-erlangen.de.
authors.forename: Markus F
authors.initials: MF
authors.lastname: Neurath
mesh_terms: D000285:Administration, Rectal; D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D016022:Case-Control Studies; D002648:Child; D003092:Colitis / Q000139:chemically induced / Q000378:metabolism* / Q000517:prevention & control*; D003093:Colitis, Ulcerative / Q000235:genetics / Q000378:metabolism; D003106:Colon / Q000737:chemistry / Q000378:metabolism; D003424:Crohn Disease / Q000235:genetics / Q000378:metabolism; D016207:Cytokines / Q000378:metabolism; D021881:DNA, Catalytic / Q000008:administration & dosage*; D004195:Disease Models, Animal; D005260:Female; D050990:GATA3 Transcription Factor / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D008875:Middle Aged; D010081:Oxazolone; D012333:RNA, Messenger / Q000032:analysis*; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics; D013601:T-Lymphocytes / Q000378:metabolism; D014302:Trinitrobenzenesulfonic Acid; D055815:Young Adult
chemical_list: D016207:Cytokines; D021881:DNA, Catalytic; D050990:GATA3 Transcription Factor; C494749:GATA3 protein, human; C494748:Gata3 protein, mouse; C583023:Hgd40 DNAzyme; D012333:RNA, Messenger; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D010081:Oxazolone; D014302:Trinitrobenzenesulfonic Acid
keywords: DNA Cleavage; Gene Regulation; Immune Response; Mouse Model