title: Optimal Arrangement of Four Short DNA Strands for Delivery of Immunostimulatory Nucleic Acids to Immune Cells.
abstract: Nanosized DNA assemblies are useful for delivering immunostimulatory cytosine-phosphate-guanine (CpG) DNA to immune cells, but little is known about the optimal structure for such delivery. In this study, we designed three different DNA nanostructures using four 55-mer oligodeoxynucleotides (ODNs), that is, tetrapod-like structured DNA (tetrapodna), tetrahedral DNA (tetrahedron), and tetragonal DNA (tetragon), and compared their potencies. Electrophoresis showed that tetrapodna was obtained with high yield and purity, whereas tetrahedron formed multimers at high ODN concentrations. Atomic force microscopy revealed that all preparations were properly constructed under optimal conditions. The thermal stability of tetrapodna was higher than those of the others. Dynamic light scattering analysis showed that all of the assemblies were about 8 nm in diameter. Upon addition to mouse macrophage-like RAW264.7 cells, tetrahedron was most efficiently taken up by the cells. Then, a CpG DNA, a ligand for toll-like receptor 9, was linked to these DNA nanostructures and added to RAW264.7 cells. CpG tetrahedron induced the largest amount of tumor necrosis factor-α, followed by CpG tetrapodna. Similar results were obtained using human peripheral blood mononuclear cells. Taken together, these results indicate that tetrapodna is the best assembly with the highest yield and high immunostimulatory activity, and tetrahedron can be another useful assembly for cellular delivery if its preparation yield is improved.
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Shozo
authors.initials: S
authors.lastname: Ohtsuki
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Matsuzaki
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Kohta
authors.initials: K
authors.lastname: Mohri
authors.affiliation: 2 Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University , Kyoto, Japan .
authors.forename: Masayuki
authors.initials: M
authors.lastname: Endo
authors.affiliation: 3 Department of Chemistry, Graduate School of Science, Kyoto University , Kyoto, Japan .
authors.forename: Tomoko
authors.initials: T
authors.lastname: Emura
authors.affiliation: 3 Department of Chemistry, Graduate School of Science, Kyoto University , Kyoto, Japan .
authors.forename: Kumi
authors.initials: K
authors.lastname: Hidaka
authors.affiliation: 2 Institute for Integrated Cell-Material Sciences (iCeMS), Kyoto University , Kyoto, Japan .|3 Department of Chemistry, Graduate School of Science, Kyoto University , Kyoto, Japan .
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Sugiyama
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Yuki
authors.initials: Y
authors.lastname: Takahashi
authors.affiliation: 4 Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University , Kyoto, Japan .
authors.forename: Kenichi
authors.initials: K
authors.lastname: Ishiyama
authors.affiliation: 4 Department of Hematology and Oncology, Graduate School of Medicine, Kyoto University , Kyoto, Japan .
authors.forename: Norimitsu
authors.initials: N
authors.lastname: Kadowaki
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Takakura
authors.affiliation: 1 Department of Biopharmaceutics and Drug Metabolism, Graduate School of Pharmaceutical Sciences, Kyoto University , Kyoto, Japan .
authors.forename: Makiya
authors.initials: M
authors.lastname: Nishikawa
mesh_terms: D000276:Adjuvants, Immunologic / Q000378:metabolism / Q000494:pharmacology*; D000818:Animals; D002738:Chloroquine / Q000494:pharmacology; D018899:CpG Islands; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D006801:Humans; D016898:Interferon-alpha / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D009690:Nucleic Acid Conformation; D000067996:RAW 264.7 Cells; D044366:Transition Temperature
chemical_list: D000276:Adjuvants, Immunologic; D004277:DNA, Single-Stranded; D016898:Interferon-alpha; D002738:Chloroquine