mPEG-PAMAM-G4 Nucleic Acid Nanocomplexes: Enhanced Stability, RNase Protection, and Activity of Splice Switching Oligomer and Poly I:C RNA

• Published in: Biomacromolecules Vol. 14; no. 11; pp. 4108 - 4115
• Main Authors: Reyes-Reveles, Juan, Sedaghat-Herati, Reza, Gilley, David R, Schaeffer, Ashley M, Ghosh, Kartik C, Greene, Thomas D, Gann, Hannah E, Dowler, Wesley A, Kramer, Stephen, Dean, John M, Delong, Robert K
• Format: Journal Article
• Language: English
• Published: Washington, DC American Chemical Society 11.11.2013
• Subjects: Alternative Splicing - genetics; Animals; Antineoplastic Agents - metabolism; Applied sciences; Biological and medical sciences; Cell Line, Tumor; Cell Survival - drug effects; Circular Dichroism; Dendrimers - chemistry; Dendrimers - pharmacology; Drug Carriers - chemistry; Drug Carriers - pharmacology; Exact sciences and technology; Female; General pharmacology; Humans; Medical sciences; Melanoma - genetics; Melanoma - pathology; Mice; Microscopy, Atomic Force; Molecular Structure; Nanostructures - chemistry; Nylons - chemistry; Nylons - pharmacology; Oligonucleotides - genetics; Oligonucleotides - metabolism; Organic polymers; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Physicochemistry of polymers; Poly I-C - genetics; Poly I-C - metabolism; Polyethylene Glycols - chemistry; Polyethylene Glycols - pharmacology; Properties and characterization; Ribonuclease, Pancreatic - metabolism; RNA - genetics; RNA - metabolism; RNA Stability - drug effects; Special properties (catalyst, reagent or carrier); Amidoacid copolymer; Antineoplastic agent; Circular dichroism spectrum; Polyplex; Dendrimer; RNA; Control release polymer; Aromatic polymer; Asymmetric molecule; Drug carrier; Branched copolymer; Ethylene oxide copolymer; Experimental study; Dimension spectrum; Biological activity; Transfection; Malignant melanoma; Diblock copolymer; Oligonucleotide; Aqueous solution; Tumor cell; Aliphatic copolymer
• ISSN: 1525-7797; 1526-4602
• DOI: 10.1021/bm4012425

Dendrimer chemistries have virtually exploded in recent years with increasing interest in this class of polymers as gene delivery vehicles. An effective nucleic acid delivery vehicle must efficiently bind its cargo and form physically stable complexes. Most importantly, the nucleic acid must be protected in biological fluids and tissues, as RNA is extremely susceptible to nuclease degradation. Here, we characterized the association of nucleic acids with generation 4 PEGylated poly(amidoamine) dendrimer (mPEG-PAMAM-G4). We investigated the formation, size, and stability over time of the nanoplexes at various N/P ratios by gel shift and dynamic light scatter spectroscopy (DLS). Further characterization of the mPEG-PAMAM-G4/nucleic acid association was provided by atomic force microscopy (AFM) and by circular dichroism (CD). Importantly, mPEG-PAMAM-G4 complexation protected RNA from treatment with RNase A, degradation in serum, and various tissue homogenates. mPEG-PAMAM-G4 complexation also significantly enhanced the functional delivery of RNA in a novel engineered human melanoma cell line with splice-switching oligonucleotides (SSOs) targeting a recombinant luciferase transcript. mPEG-PAMAM-G4 triconjugates formed between gold nanoparticle (GNP) and particularly manganese oxide (MnO) nanorods, poly IC, an anticancer RNA, showed enhanced cancer-killing activity by an MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay.