title: Guanine modification of inhibitory oligonucleotides potentiates their suppressive function.
abstract: Inhibitory TLR7 and/or TLR9 oligonucleotides (inhibitory oligonucleotide [INH-ODN]) are characterized by a phosphorothioate backbone and a CC(T)XXX₃₋₅GGG motif, respectively. INH-ODN 2088 is a prototypic member of this class of INH-ODN and acts as a TLR7 and TLR9 antagonist. It contains a G quadruple that leads to higher order structures by the formation of G tetrads. These structures are unfavorable for the prediction of their pharmacological and immunological behavior. We show in this study that modification of Gs within the G quadruple by 7-deaza-guanine or 7-deaza-2'-O-methyl-guanine avoids higher order structures and improves their inhibitory potential. Whereas TLR9-induced TNF-α secretion of bone marrow-derived macrophages and conventional dendritic cells was equally inhibited by INH-ODN 2088 and G-modified INH-ODNs such as INH-ODN 24888, TLR7-induced TNF-α release and TLR7- and TLR9-induced IL-12p40 release were significantly more impaired by G-modified INH-ODNs. Similarly, the IL-6 release of B cells from wild-type and autoimmune MRL/Mp-lpr/lpr mice was more efficiently impaired by G-modified INH-ODNs. Surprisingly, INH-ODN 2088 stimulated B cells to proliferate when used in higher doses. Finally, in vivo, in wild-type and autoimmune MRL/Mp-lpr/lpr mice, G-modified INH-ODN 24888 was significantly more efficient than unmodified INH-ODN 2088. In summary, G modification allows the development of INH-ODNs with superior inhibitory potency for inflammatory diseases with high medical need such as systemic lupus erythematosus.
authors: Institut für Medizinische Mikrobiologie, Immunologie, und Hygiene, Technische Universität München, 81675 Munich, Germany.
authors: Franziska
authors: F
authors: Römmler
authors: Marion
authors: M
authors: Jurk
authors: Eugen
authors: E
authors: Uhlmann
authors: Monika
authors: M
authors: Hammel
authors: Anna
authors: A
authors: Waldhuber
authors: Lavinia
authors: L
authors: Pfeiffer
authors: Hermann
authors: H
authors: Wagner
authors: Jörg
authors: J
authors: Vollmer
authors: Thomas
authors: T
authors: Miethke
mesh_terms: D000818:Animals; D001402:B-Lymphocytes / Q000187:drug effects / Q000276:immunology; D049109:Cell Proliferation / Q000187:drug effects; D016207:Cytokines / Q000096:biosynthesis; D003713:Dendritic Cells / Q000187:drug effects / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D006147:Guanine / Q000737:chemistry*; D007108:Immune Tolerance / Q000187:drug effects* / Q000276:immunology; D008264:Macrophages / Q000187:drug effects / Q000276:immunology; D008562:Membrane Glycoproteins / Q000037:antagonists & inhibitors; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D019463:Mice, Inbred MRL lpr; D009841:Oligonucleotides / Q000737:chemistry* / Q000494:pharmacology*; D051199:Toll-Like Receptor 7 / Q000037:antagonists & inhibitors; D051217:Toll-Like Receptor 9 / Q000037:antagonists & inhibitors
chemical_list: D016207:Cytokines; D008562:Membrane Glycoproteins; D009841:Oligonucleotides; C477038:Tlr7 protein, mouse; C495370:Tlr9 protein, mouse; D051199:Toll-Like Receptor 7; D051217:Toll-Like Receptor 9; D006147:Guanine