title: Absence of unspecific innate immune cell activation by GATA-3-specific DNAzymes.
abstract: DNAzymes of the 10-23 family represent an important class of antisense molecules with implications for therapeutic treatment of diseases. These molecules are single-stranded oligodeoxynucleotides combining the high specificity of oligonucleotide base pairing with an inherent RNA-cleaving enzymatic activity. However, like other oligonucleotide-based molecules these substances might exert so-called off-target effects, which have not been investigated so far for this molecule class. Therefore, the present study investigates putative off-target effects of DNAzymes on innate immune mechanisms using GATA-3-specific DNAzymes that have recently been developed as novel therapeutic approach for the treatment of allergic diseases including allergic asthma. The conserved catalytic domain of 10-23 DNAzymes contains a CpG motif that may stimulate innate immune cells via Toll-like receptor 9 (TLR-9). Therefore, potential TLR-9-mediated as well as TLR-9 independent cell activation was investigated using TLR-9-transfected HEK293 cells, macrophage cell lines and primary innate immune cells. Furthermore, putative effects of GATA-3-specific DNAzymes on the activation of neutrophil granulocytes and degranulation of mast cells/basophils were analyzed. In summary, no innate immune cell-stimulating activities of the tested DNAzymes were observed in any of the systems. Consequently, use of GATA-3-specific DNAzymes may represent a novel and highly specific approach for the treatment of allergic diseases.
authors.affiliation: Sterna Biologicals GmbH & Co. KG, Marburg, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Dicke
authors.forename: Isabella
authors.initials: I
authors.lastname: Pali-Schöll
authors.forename: Andreas
authors.initials: A
authors.lastname: Kaufmann
authors.forename: Stefan
authors.initials: S
authors.lastname: Bauer
authors.forename: Harald
authors.initials: H
authors.lastname: Renz
authors.forename: Holger
authors.initials: H
authors.lastname: Garn
mesh_terms: D000818:Animals; D001483:Base Sequence; D001491:Basophils / Q000187:drug effects / Q000378:metabolism; D020134:Catalytic Domain; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D021881:DNA, Catalytic / Q000494:pharmacology*; D004277:DNA, Single-Stranded / Q000494:pharmacology*; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D050990:GATA3 Transcription Factor; D017930:Genes, Reporter; D007107:Immune System / Q000166:cytology*; D007113:Immunity, Innate / Q000187:drug effects*; D007155:Immunologic Factors / Q000494:pharmacology*; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D008264:Macrophages / Q000187:drug effects / Q000276:immunology / Q000378:metabolism; D008407:Mast Cells / Q000187:drug effects / Q000201:enzymology; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000235:genetics; D009504:Neutrophils / Q000187:drug effects / Q000378:metabolism; D061251:Primary Cell Culture; D051381:Rats; D016897:Respiratory Burst / Q000187:drug effects; D013154:Spleen / Q000166:cytology; D051217:Toll-Like Receptor 9 / Q000378:metabolism
chemical_list: D016207:Cytokines; D021881:DNA, Catalytic; D004277:DNA, Single-Stranded; D050990:GATA3 Transcription Factor; D007155:Immunologic Factors; D016328:NF-kappa B; C402582:RNA-cleaving DNA 10-23; C495370:Tlr9 protein, mouse; D051217:Toll-Like Receptor 9; D008156:Luciferases