title: A DNA microarray-based analysis of immune-stimulatory and transcriptional responses of dendritic cells to KALA-modified nanoparticles.
abstract: Technologies for the transfection of antigen-encoding genes into the dendritic cells, and subsequent immune-activation are both prerequisites for a successful DNA vaccine. We herein report on the density-dependent enhancement of transgene expression by the simple modification by stearyl-conjugated KALA, an α-helical peptide (STR-KALA), onto a lipid envelope-type nanoparticle (the R8-MEND, an octaarginine-modified multifunctional envelope-type nano device). The enhanced transgene expression in the KALA-modified R8-MEND (R8/KALA-MEND) cannot be explained by cellular uptake and nuclear delivery efficacy. Thus, the post-nuclear delivery process (i.e. transcription), but not intracellular trafficking processes attributed the enhanced transfection efficacy. Microarray analyses revealed that transfection with the R8/KALA-MEND resulted in a greater perturbation in host genes expression in comparison with the R8-MEND and that this effect was time-dependent. Further pathway analyses in the category of transcription-related genes and a gene ontology analysis indicated that the R8/KALA-MEND stimulated the expression of transcription factors that are closely related to immune-activation (i.e. NF-kB and STAT). Inhibition of the transfection efficacy by blockage of the STAT pathways revealed that the enhanced transcription activity is the result of immune-stimulation. Collectively, the R8/KALA-MEND mounts a "switch-on" function that triggers signal transduction forward to the immune-stimulation analogous to an adjuvant, and consequently elicits active transcription.
authors: Laboratory for Molecular Design of Pharmaceutics, Faculty of Pharmaceutical Sciences, Hokkaido University, Kita-12, Nishi-6, Kita-ku, Sapporo 060-0812, Japan. akita@pharm.hokudai.ac.jp
authors: Hidetaka
authors: H
authors: Akita
authors: Soichiro
authors: S
authors: Ishii
authors: Naoya
authors: N
authors: Miura
authors: Sharif Mohammad
authors: SM
authors: Shaheen
authors: Yasuhiro
authors: Y
authors: Hayashi
authors: Takashi
authors: T
authors: Nakamura
authors: Noritada
authors: N
authors: Kaji
authors: Yoshinobu
authors: Y
authors: Baba
authors: Hideyoshi
authors: H
authors: Harashima
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000737:chemistry / Q000187:drug effects; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D005260:Female; D018014:Gene Transfer Techniques; D007113:Immunity, Innate; D016899:Interferon-beta / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D053758:Nanoparticles / Q000737:chemistry*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D009842:Oligopeptides / Q000737:chemistry; D015398:Signal Transduction; D014162:Transfection; D019076:Transgenes; D019444:Vaccines, DNA / Q000737:chemistry
chemical_list: D016328:NF-kappa B; D009842:Oligopeptides; D019444:Vaccines, DNA; C448619:octaarginine; D016899:Interferon-beta
keywords: Adjuvant; DNA vaccine; Innate immunity; Microarray; Nanoparticle