title: A peptide-based dendrimer that enhances the splice-redirecting activity of PNA conjugates in cells.
abstract: The full therapeutic potential of oligonucleotide (ON)-based agents has been hampered by cellular delivery challenges. Cell-penetrating peptides (CPP) represent promising delivery vectors for nucleic acids, and their potential has recently been evaluated using a functional splicing redirection assay, which capitalizes on the nuclear delivery of splice-correcting steric-block ON analogues such as peptide nucleic acids (PNA). Despite encouraging in vitro and in vivo data with arginine-rich CPP-steric block conjugates, mechanistic studies have shown that entrapment within the endosome/lysosome compartment after endocytosis remains a limiting factor. Previous work from our group has shown that CPP oligomerization greatly improves cellular delivery and increases transfection of plasmid DNA. We now report the chemical synthesis and the evaluation of multivalent CPP-PNA constructs incorporating monomeric (p53(mono)) and dendrimer-like tetrameric (p53(tet)) forms of the p53 tetramerization domain containing peptide, a 10 arginine CPP domain (R10), and a splice redirecting PNA (PNA705). These CPP-PNA conjugates were termed R10p53(tet)-PNA705 and R10p53(mono)-PNA705, referring to their oligomerization state. The present study demonstrates that the splicing redirection efficiency of PNA705 is much greater in the context of the tetrameric R10p53(tet)-PNA705 construct than for the monomeric and occurs at nanomolar concentrations, demonstrating that multivalency is an important factor in delivering PNA into cells.
authors: UMR 5235 CNRS, Université Montpellier 2, Montpellier, France.
authors: Fatouma Said
authors: FS
authors: Hassane
authors: Gabriela D
authors: GD
authors: Ivanova
authors: Eleonora
authors: E
authors: Bolewska-Pedyczak
authors: Rachida
authors: R
authors: Abes
authors: Andrey A
authors: AA
authors: Arzumanov
authors: Michael J
authors: MJ
authors: Gait
authors: Bernard
authors: B
authors: Lebleu
authors: Jean
authors: J
authors: Gariépy
mesh_terms: D017398:Alternative Splicing / Q000187:drug effects*; D057846:Cell-Penetrating Peptides / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D050091:Dendrimers / Q000138:chemical synthesis / Q000737:chemistry* / Q000494:pharmacology*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006367:HeLa Cells; D006801:Humans; D020135:Peptide Nucleic Acids / Q000138:chemical synthesis / Q000737:chemistry* / Q000235:genetics / Q000494:pharmacology*
chemical_list: D057846:Cell-Penetrating Peptides; D050091:Dendrimers; D020135:Peptide Nucleic Acids