Original ArticleSelf-assembling DNA hydrogel-based delivery of immunoinhibitory nucleic acids to immune cells
Graphical Abstract
Takumi-shaped DNA consisting of two partially complementary immunoinhibitory oligodeoxynucleotides and Takumi-based DNA hydrogels were developed as a useful delivery vehicle for the oligodeoxynucleotides to immune cells for the inhibition of Toll-like receptor 9-mediated hyperinduction of proinflammatory cytokines.
Section snippets
Chemicals
RPMI1640 medium was obtained from Nissui Pharmaceutical Co., Ltd. (Tokyo, Japan). Fetal bovine serum (FBS) was obtained from Hyclone (South Logan, UT, USA). Opti-modified Eagle’s medium (Opti-MEM) and MEM non-essential amino acids solution (NEAA) were purchased from Life Technologies (Carlsbad, CA, USA). Monothioglycerol was obtained from Wako Pure Chemical (Osaka, Japan). The 20-bp ladder was purchased from Takara Bio, Inc. (Shiga, Japan). Propidium iodide (PI) and blue dextran (average
Formation of Takumi and DNA hydrogel
Figure 1 shows the putative structures of the Takumi and Takumi-based DNA hydrogel. Two ODNs each were used to prepare the Takumi (iODN1 and iODN2) and DNA hydrogel (iODN1 and iODN3). The Takumi for the DNA hydrogel was designed such that both the 5′-ends and the 3′-ends are complementary to each other. Since these Takumi and DNA hydrogels contain INH-ODNs, we called them inhibitory Takumi (iTakumi) and inhibitory DNA hydrogel (iTakumiGel), respectively. Figure 2, A shows the PAGE analysis of
Discussion
Our previous studies demonstrated that the organization of CpG DNA into a branched form significantly increases its immunostimulatory activity.15, 16 In this study, we created a branched DNA, called Takumi, using two 33-mer ODNs, which was used as the main component of a DNA hydrogel. The AFM image clearly showed that the branched DNA adopted a Takumi shape as designed (Figure 2, B), which supports the notion that the smaller iTakumi units are also in a Takumi shape. An important finding of
Acknowledgment
The authors would like to thank Prof. Hiroshi Watanabe of Institute for Chemical Research, Kyoto University, for his helpful advice and discussion on the rheological properties of the samples.
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2021, Advanced Drug Delivery ReviewsCitation Excerpt :On the other hand, the delivery of immunoinhibitory oligodeoxynucleotides (INH-ODNs) has shown efficacy with autoimmune diseases associated with Toll-like receptor 9 (TLR9) activation [142,143]. Nishikawa and coworkers synthesized a DNA hydrogel with a Takumi-shaped DNA branch consisting of two partially complementary INH-ODNs [144]. This hydrogel promoted the cellular uptake of ODNs and hindered TLR9-mediated cytokine release.
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The authors have no conflict of interest and no disclosures.
This work was supported in part by a Grant-in-Aid for Scientific Research (B) (23390010) from the Japan Society for the Promotion of Science, by a Grant-in-Aid for Scientific Research on Innovative Areas “Carcinogenic spiral” (25114706) from theMinistry of Education, Culture, Sports, Science, and Technology of Japan, and by a Cross-Disciplinary Collaborative Research Promotion Project from the Institute for Integrated Cell-Material Sciences, Kyoto University.