Characterization of biomolecular nanoconjugates by high-throughput delivery and spectroscopic difference
Abstract
Aim: Nanoparticle conjugates have the potential for delivering siRNA, splice-shifting oligomers or nucleic acid vaccines, and can be applicable to anticancer therapeutics. This article compares tripartite conjugates with gold nanoparticles or synthetic methoxypoly(ethylene glycol)-block-polyamidoamine dendrimers. Materials & methods: Interactions with model liposomes of a 1:1 molar ratio of tripalmitin:cholesterol or phospholipid:cholesterol were investigated by high-throughput absorbance, as well as fluorescence difference and cellular luminescence assays. Results: Spectral differences and dynamic light-scattering spectroscopy shifts demonstrated the interaction of conjugates with liposomes. Biological activity was demonstrated by upregulation of gene expression via splice-shifting oligomers, delivery of anti-B-Raf siRNA in cultured human cancer cells or tuberculosis antigen 85B plasmid expression vector in a coculture model of antigen presentation. Conclusion: The data suggests that gold nanoparticles and methoxypoly(ethylene glycol)-block-polyamidoamine dendrimer nanoconjugates may have potential for binding, stabilization and delivery of splice-shifting oligomers, siRNA and nucleic acid vaccines for preclinical trials.
Original submitted 19 December 2011; Revised submitted 3 April 2012; Published online 3 September 2012
Papers of special note have been highlighted as: ▪ of interest ▪▪ of considerable interest
References
- 1 Bárány-Wallje E, Keller S, Serowy S et al. A critical reassessment of penetratin translocation across lipid membranes. Biophys. J.89(4),2513–2521 (2005).Crossref, Medline, CAS, Google Scholar
- 2 Shoemaker RH, Scudiero DA, Melillo G et al. Application of high-throughput, molecular-targeted screening to anticancer drug discovery. Curr. Top. Med. Chem.2(3),229–246 (2002).Crossref, Medline, CAS, Google Scholar
- 3 Chithrani DB. Intracellular uptake, transport and processing of gold nanostructures. Mol. Membr. Biol.27(7),299–311 (2010).Crossref, Medline, CAS, Google Scholar
- 4 Vijayanathan V, Thomas T, Thomas TJ. DNA nanoparticles and development of DNA delivery vehicles for gene therapy. Biochemistry41(48),14085–14094 (2002).▪▪ Describes the importance of polyamines and polymers as DNA delivery vehicles.Crossref, Medline, CAS, Google Scholar
- 5 Brewer LR, Corzett M, Balhorn R. Protamine-induced condensation and decondensation of the same DNA molecule. Science286(5437),120–123 (1999).Crossref, Medline, CAS, Google Scholar
- 6 Reynolds F, Weissleder R, Josephson L. Protamine as an efficient membrane-translocating peptide. Bioconjug. Chem.16(5),1240–1245 (2005).Crossref, Medline, CAS, Google Scholar
- 7 Cheng Y, Xu Z, Ma M, Xu T. Dendrimers as drug carriers: applications in different routes of drug administration. J. Pharm. Sci.97(1),123–143 (2008).Crossref, Medline, CAS, Google Scholar
- 8 Veronese FM, Pasut G. PEGylation, successful approach to drug delivery. Drug Discov. Today10(21),1451–1458 (2005).Crossref, Medline, CAS, Google Scholar
- 9 Froehlich E, Mandeville JS, Jennings, CJ, Segadhat-Herati R, Tajmir-Riahi HA. Dendrimers bind human serum albumin. J. Phys. Chem. B113(19),6986–6993 (2009).▪ Supports the authors’ reasoning for using generation 3 methoxypoly(ethylene glycol)-block-polyamidoamine dendrimer in their studies.Crossref, Medline, CAS, Google Scholar
- 10 Davis ME, Zuckerman JE, Choi CHJ et al. Evidence of RNAi in humans from systemically administered siRNA via targeted nanoparticles. Nature464(7291),1067–1070 (2010).Crossref, Medline, CAS, Google Scholar
- 11 Kutzler MA Weiner DB. DNA vaccines: ready for the prime time. Nat. Gen.9(10),776–788 (2008).Crossref, CAS, Google Scholar
- 12 Navabi H, Jasani B, Reece A et al. A clinical grade poly I:C-analogue (Ampligen) promotes optimal DC maturation and Th1-type T cell responses of healthy donors and cancer patients in vitro. Vaccine27(1),107–115 (2009).Crossref, Medline, CAS, Google Scholar
- 13 Liu MA. DNA vaccines: a historical perspective and view to the future. Immunol. Rev.239(1),62–84 (2011).Crossref, Medline, CAS, Google Scholar
- 14 Bauman JA, Li SD, Yang A, Huang L, Kole R. Anti-tumor activity of splice-switching oligonucleotides. Nucleic Acids Res.38(22),8348–8356 (2010).Crossref, Medline, CAS, Google Scholar
- 15 Garcia-Blanco MA, Baraniak AP, Lasda EL. Alternative splicing in disease and therapy. Nat. Biotechnol.22(5),535–546 (2004).Crossref, Medline, CAS, Google Scholar
- 16 Hingorani SR, Jacobetz MA, Robertson GP, Herlyn M, Tuveson DA. Suppression of BRAF(V599E) in human melanoma abrogates transformation. Cancer Res.63(17),5198–5202 (2003).Medline, CAS, Google Scholar
- 17 Aagaard C, Hoang TT, Izzo A et al. Protection and polyfunctional T cells induced by Ag85B-TB10.4/IC31 against Mycobacterium tuberculosis is highly dependent on the antigen dose. PLoS ONE4(6),e5930 (2009).Crossref, Medline, Google Scholar
- 18 DeLong RK, Akhtar U, Sallee M et al. Characterization and performance of nucleic acid nanoparticles combined with protamine and gold. Biomaterials30(32),6451–6459 (2009).▪▪ Former research carried out by our group, which underlies the basis of the current study.Crossref, Medline, CAS, Google Scholar
- 19 DeLong RK, Cillessen L, Reynolds C et al. Biomolecular triconjugates formed between gold, protamine and nucleic acid: comparative characterization on the nanoscale. J. Nanotechnol. doi:10.1155/2012/954601 (2012) (Epub ahead of print).Google Scholar
- 20 Arvizo RR, Miranda OR, Thompson MA et al. Effect of nanoparticle surface charge at the plasma membrane and beyond. Nano Lett.10(7),2543–2548 (2010).Crossref, Medline, CAS, Google Scholar
- 21 Iyer J, Fleming K, Hammond PT. Synthesis and solution properties of new linear-dendritic diblock copolymers. Macromolecules31,8757–8765 (1998).Crossref, CAS, Google Scholar
- 22 Ramachandra L, Smialek JL, Shank SS et al. Phagosomal processing of Mycobacterium tuberculosis Antigen85B is modulated independently of mycobacterial viability and phagosome maturation. Infect. Immun.73(2),1097–1105 (2005).Crossref, Medline, CAS, Google Scholar
- 23 Sharma A, Desai A, Ali R, Tomalia D. Polyacrylamide gel electrophoresis separation and detection of polyamidoamine dendrimers possessing various cores and terminal groups. J. Chromatogr A.1081(2),238–244 (2005).Crossref, Medline, CAS, Google Scholar
- 24 Luo D, Haverstick K, Belcheva N, Han E, Saltzman WM. Poly(ethylene glycol)-conjugated PAMAM dendrimer for biocompatible, high-efficiency DNA delivery. Macromolecules35(9),3456–3462 (2002).Crossref, CAS, Google Scholar
- 25 Chittimalla C, Zammut-Italiano L, Zuber G, Behr JP. Monomolecular DNA nanoparticles for intravenous delivery of genes. J. Am. Chem. Soc.127(32),11436–11441 (2005).Crossref, Medline, CAS, Google Scholar
- 26 Tombaccini D, Fallani A, Mugnai G, Ruggieri S. Lipid composition of Balb/c3T3, SV3T3, and concanavalin A-selected revertant cells grown in media containing lipid-depleted serum. J. Lip. Res.22(4),590–597 (1981).Crossref, Medline, CAS, Google Scholar
