title: A multifunctional PEI-based cationic polyplex for enhanced systemic p53-mediated gene therapy.
abstract: We recently reported a novel coupling strategy involving salicylhydroxamic acid and phenyl(di)boronic acid molecules to attach the CNGRC peptide to PEI/DNA for CD13 targeting in tumors. Here, we doubly coupled Simian Virus (SV) 40 peptide-(nuclear localization signal)) and oligonucleotide-based (DNA nuclear targeting signal) nuclear signals to the same vector using peptide nucleic acid chemistry. This vector, CNGRC/PEG/PEI/DNA-betagal/NLS/DNTS, was predominantly localized in the cell nucleus, yielding about 200-fold higher betagal gene expression in vitro, more than 20-fold increase in tumor-specific gene delivery, and a robust betagal gene expression as demonstrated in stained tumor sections. For gene therapy purposes, we further engineered a similar targeting polyplex, CNGRC/PEG/PEI/DNA-p53/NLS/DNTS, with EBV-based episomal vector for sustained p53 gene expression. A distribution of vector DNA and apoptosis in p53-containing tumors was observed, yielding a significant tumor regression and 95% animal survival after 60 days. This multicomponent vector also co-targeted tumor and tumor-associated endothelial cells but not normal cells, and had more efficient therapeutic index than each vector administered as a single modality. The use of an efficient coupling strategy without compromising the vector's integrity for DNA condensation and endosomal escape; nuclear import; tumor-specific and persistent p53 gene expression clearly provides a basis for developing a single combinatorial approach for non-viral gene therapy.
authors.affiliation: Department of Genitourinary Medical Oncology, University of Texas MD Anderson Cancer Center, 1515 Holcombe Boulevard, Houston, TX 77030, USA. stanley-moffatt@excite.com
authors.forename: S
authors.initials: S
authors.lastname: Moffatt
authors.forename: S
authors.initials: S
authors.lastname: Wiehle
authors.forename: R J
authors.initials: RJ
authors.lastname: Cristiano
mesh_terms: D000818:Animals; D018826:CD13 Antigens / Q000235:genetics; D002289:Carcinoma, Non-Small-Cell Lung / Q000276:immunology / Q000628:therapy*; D002412:Cations; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D004247:DNA / Q000008:administration & dosage; D015870:Gene Expression; D018390:Gene Targeting; D016158:Genes, p53; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D020287:In Situ Nick-End Labeling; D008175:Lung Neoplasms / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D011094:Polyethyleneimine; D013539:Simian virus 40 / Q000235:genetics; D014162:Transfection / Q000379:methods
chemical_list: D002412:Cations; D011094:Polyethyleneimine; D004247:DNA; D018826:CD13 Antigens