Optimizing CpG spatial distribution with DNA origami for Th1-polarized therapeutic vaccination

Multivalent presentation of ligands often enhances receptor activation and downstream signaling. DNA origami offers precise nanoscale spacing of ligands, a potentially useful feature for therapeutic nanoparticles. Here we introduce a “square block” DNA origami platform to explore the importance of spacing of CpG oligonucleotides, which engage Toll-like receptors and thereby act as danger signals for dendritic cells. Through in vitro cell-culture studies and in vivo tumor-treatment models, we demonstrate that square blocks induce Th1 immune polarization when CpG is spaced at 3.5 nm. We observe that this DNA origami vaccine enhances DC activation, antigen cross-presentation, CD8 T cell activation, Th1-polarized CD4 activation and NK cell activation. The vaccine also synergizes effectively with anti-PD-L1 for improved cancer immunotherapy in melanoma and lymphoma models and induces long-term T cell memory. Our results suggest that DNA origami may serve as an advanced vaccine platform for controlling adjuvant spacing and co-delivering antigens. One Sentence Summary This study developed a DNA origami-based cancer vaccine (DoriVac) that co-delivers antigen and CpG immune adjuvant with an optimal spacing for Th1 immune polarization.

[1]  Patrick D. Halley,et al.  DNA Origami Nanostructures Elicit Dose-Dependent Immunogenicity and Are Nontoxic up to High Doses In Vivo. , 2022, Small.

[2]  I. Parish,et al.  Spatially Controlled Activation of Toll-like Receptor 9 with DNA-Based Nanomaterials , 2022, Nano letters.

[3]  Chi Zhang,et al.  CpG Oligodeoxynucleotides for Anticancer Monotherapy from Preclinical Stages to Clinical Trials , 2021, Pharmaceutics.

[4]  Wendy Xueyi Wang,et al.  Engineering DNA Nanostructures to Manipulate Immune Receptor Signaling and Immune Cell Fates , 2021, Advanced healthcare materials.

[5]  Zhiwei Xu,et al.  CpG-Based Nanovaccines for Cancer Immunotherapy , 2021, International journal of nanomedicine.

[6]  Johann M. Weck,et al.  Nanoscale FasL Organization on DNA Origami to Decipher Apoptosis Signal Activation in Cells. , 2021, Small.

[7]  Shawn M. Douglas,et al.  Tight nanoscale clustering of Fcγ-receptors using DNA origami promotes phagocytosis , 2021, bioRxiv.

[8]  Xueliang Wang,et al.  Mannose-Modified Liposome Co-Delivery of Human Papillomavirus Type 16 E7 Peptide and CpG Oligodeoxynucleotide Adjuvant Enhances Antitumor Activity Against Established Large TC-1 Grafted Tumors in Mice , 2020, International journal of nanomedicine.

[9]  M. Röcken,et al.  The role of dendritic cells for therapy of B-cell lymphoma with immune checkpoint inhibitors , 2020, Cancer Immunology, Immunotherapy.

[10]  Baoquan Ding,et al.  A DNA nanodevice-based vaccine for cancer immunotherapy , 2020, Nature Materials.

[11]  K. Ikebukuro,et al.  Monomeric G-Quadruplex-Based CpG Oligodeoxynucleotides as Potent Toll-Like Receptor 9 Agonists. , 2020, Biomacromolecules.

[12]  Björn Högberg,et al.  Spatial organization-dependent EphA2 transcriptional responses revealed by ligand nanocalipers , 2020, Nucleic acids research.

[13]  Mark Bathe,et al.  Role of nanoscale antigen organization on B-cell activation probed using DNA origami , 2020, bioRxiv.

[14]  William M. Shih,et al.  Glutaraldehyde crosslinking of oligolysines coating DNA origami greatly reduces susceptibility to nuclease degradation. , 2020, Journal of the American Chemical Society.

[15]  Jie Chao,et al.  Designer DNA architecture offers precise and multivalent spatial pattern-recognition for viral sensing and inhibition , 2019, bioRxiv.

[16]  Björn Högberg,et al.  Binding to Nanopatterned Antigens is Dominated by the Spatial Tolerance of Antibodies , 2018, Nature Nanotechnology.

[17]  Ö. Türeci,et al.  Personalized vaccines for cancer immunotherapy , 2018, Science.

[18]  R. Vijay,et al.  Interleukin‐10 Directly Inhibits CD8+ T Cell Function by Enhancing N‐Glycan Branching to Decrease Antigen Sensitivity , 2018, Immunity.

[19]  Baoquan Ding,et al.  A DNA nanorobot functions as a cancer therapeutic in response to a molecular trigger in vivo , 2018, Nature Biotechnology.

[20]  Aileen W. Li,et al.  A facile approach to enhance antigen response for personalized cancer vaccination , 2018, Nature Materials.

[21]  A. Zloza,et al.  Nanoparticles: augmenting tumor antigen presentation for vaccine and immunotherapy treatments of cancer , 2017, Nanomedicine.

[22]  Anuttara Udomprasert,et al.  DNA origami applications in cancer therapy , 2017, Cancer science.

[23]  Baoquan Ding,et al.  Self-Assembled DNA Dendrimer Nanoparticle for Efficient Delivery of Immunostimulatory CpG Motifs. , 2017, ACS applied materials & interfaces.

[24]  David J. Mooney,et al.  Oligolysine-based coating protects DNA nanostructures from low-salt denaturation and nuclease degradation , 2017, Nature Communications.

[25]  Tian Zhang,et al.  Antigen-capturing nanoparticles improve the abscopal effect and cancer immunotherapy , 2017, Nature Nanotechnology.

[26]  K. Roy,et al.  Biophysical Attributes of CpG Presentation Control TLR9 Signaling to Differentially Polarize Systemic Immune Responses. , 2017, Cell reports.

[27]  Takashi Sato,et al.  Use of nanoparticles to deliver immunomodulatory oligonucleotides. , 2016, Wiley interdisciplinary reviews. Nanomedicine and nanobiotechnology.

[28]  Ernest Y Lee,et al.  A review of immune amplification via ligand clustering by self-assembled liquid-crystalline DNA complexes. , 2016, Advances in colloid and interface science.

[29]  Vinit Kumar,et al.  DNA Nanotechnology for Cancer Therapy , 2016, Theranostics.

[30]  Sun‐Young Kim,et al.  Sequential delivery of an anticancer drug and combined immunomodulatory nanoparticles for efficient chemoimmunotherapy , 2015, International journal of nanomedicine.

[31]  Gerard C. L. Wong,et al.  Liquid-crystalline ordering of antimicrobial peptide-DNA complexes controls TLR9 activation. , 2015, Nature materials.

[32]  D. Klinman,et al.  CpG Oligonucleotides as Cancer Vaccine Adjuvants , 2015, Vaccines.

[33]  J. Castle,et al.  Mutant MHC class II epitopes drive therapeutic immune responses to cancer , 2015, Nature.

[34]  W. Shih.  Exploiting weak interactions in DNA self-assembly , 2015, Science.

[35]  Toshiyuki Shimizu,et al.  Structural basis of CpG and inhibitory DNA recognition by Toll-like receptor 9 , 2015, Nature.

[36]  William M. Shih,et al.  Addressing the Instability of DNA Nanostructures in Tissue Culture , 2014, ACS nano.

[37]  Björn Högberg,et al.  Spatial control of membrane receptor function using ligand nanocalipers , 2014, Nature Methods.

[38]  W. Heath,et al.  DEC-205 is a cell surface receptor for CpG oligonucleotides , 2013, Oncoimmunology.

[39]  P. Hermonat,et al.  AAV2/IL-12 gene delivery into dendritic cells (DC) enhances CTL stimulation above other IL-12 applications , 2012, Oncoimmunology.

[40]  Stefan Bauer,et al.  TLR13 Recognizes Bacterial 23S rRNA Devoid of Erythromycin Resistance–Forming Modification , 2012, Science.

[41]  Andrea I. McClatchey,et al.  Spatial regulation of receptor tyrosine kinases in development and cancer , 2012, Nature Reviews Cancer.

[42]  C. Davies,et al.  CD205 antigen targeting combined with dendritic cell recruitment factors and antigen-linked CD40L activation primes and expands significant antigen-specific antibody and CD4(+) T cell responses following DNA vaccination of outbred animals. , 2012, Vaccine.

[43]  K. Gothelf,et al.  Multilayer DNA origami packed on hexagonal and hybrid lattices. , 2012, Journal of the American Chemical Society.

[44]  P. Hermonat,et al.  Evidence for IL-12 intracrine activity in DC , 2012 .

[45]  Tim Liedl,et al.  Cellular immunostimulation by CpG-sequence-coated DNA origami structures. , 2011, ACS nano.

[46]  L. I. Bouwman,et al.  Chicken TLR21 Is an Innate CpG DNA Receptor Distinct from Mammalian TLR9 , 2010, The Journal of Immunology.

[47]  D. Ingber,et al.  Self-assembly of three-dimensional prestressed tensegrity structures from DNA , 2010 .

[48]  Shawn M. Douglas,et al.  Folding DNA into Twisted and Curved Nanoscale Shapes , 2009, Science.

[49]  Adam H. Marblestone,et al.  Rapid prototyping of 3D DNA-origami shapes with caDNAno , 2009, Nucleic acids research.

[50]  Shawn M. Douglas,et al.  Self-assembly of DNA into nanoscale three-dimensional shapes , 2009, Nature.

[51]  K. Sakurai,et al.  Enhanced cytokine secretion owing to multiple CpG side chains of DNA duplex. , 2008, Oligonucleotides.

[52]  L. Coussens,et al.  Polarized immune responses differentially regulate cancer development , 2008, Immunological reviews.

[53]  Shawn M. Douglas,et al.  DNA-nanotube-induced alignment of membrane proteins for NMR structure determination , 2007, Proceedings of the National Academy of Sciences.

[54]  S. Antonia,et al.  Surgical excision combined with autologous whole tumor cell vaccination is an effective therapy for murine neuroblastoma. , 2006, Journal of pediatric surgery.

[55]  P. Rothemund.  Folding DNA to create nanoscale shapes and patterns , 2006, Nature.

[56]  Bali Pulendran,et al.  Translating Innate Immunity into Immunological Memory: Implications for Vaccine Development , 2006, Cell.

[57]  P. Cresswell,et al.  Enhanced and prolonged cross‐presentation following endosomal escape of exogenous antigens encapsulated in biodegradable nanoparticles , 2006, Immunology.

[58]  F. Pijpers,et al.  Therapeutic cancer vaccines , 2005, Nature Reviews Drug Discovery.

[59]  D. Busch,et al.  Vaccination with Plasmid DNA Activates Dendritic Cells via Toll-Like Receptor 9 (TLR9) but Functions in TLR9-Deficient Mice 1 , 2003, The Journal of Immunology.

[60]  D. Klinman.  CpG DNA as a vaccine adjuvant , 2003, Expert review of vaccines.

[61]  G. Hartmann,et al.  Combined dendritic cell‐ and CpG oligonucleotide‐based immune therapy cures large murine tumors that resist chemotherapy , 2002, European journal of immunology.

[62]  Dong Yu,et al.  'Immunomers'--novel 3'-3'-linked CpG oligodeoxyribonucleotides as potent immunomodulatory agents. , 2002, Nucleic acids research.

[63]  R. Scheule,et al.  CpG-depleted plasmid DNA vectors with enhanced safety and long-term gene expression in vivo. , 2002, Molecular therapy : the journal of the American Society of Gene Therapy.

[64]  S. Akira,et al.  Human TLR9 confers responsiveness to bacterial DNA via species-specific CpG motif recognition , 2001, Proceedings of the National Academy of Sciences of the United States of America.