title: Heat shock protein-mediated cell penetration and cytosolic delivery of macromolecules by a telomerase-derived peptide vaccine.
abstract: A reverse-transcriptase-subunit of telomerase (hTERT) derived peptide, GV1001, has been developed as a vaccine against various cancers. Here, we report an unexpected function of GV1001 as a cell-penetrating peptide (CPP). GV1001 was delivered into a variety of cells including various cancer cell lines and primary blood cells. Moreover, the delivered GV1001 was predominantly located in the cytoplasm of the cells, while a significantly higher proportion of TAT peptide was localized in the nucleus. Macromolecules such as proteins, DNA and siRNA, which were linked to GV1001 by direct covalent conjugation or non-covalent complexation through poly-lysine, were successfully delivered into cells, indicating that GV1001 can be used as a carrier for macromolecules. Expression of the delivered DNA, and lowered expression of the target gene by the delivered siRNA, suggest the potential therapeutic use of GV1001. Pull-down analysis identified Heat Shock Protein 90 (HSP90) and 70 (HSP70) as GV1001 interacting proteins. Treatment of Anti-HSP90 and HSP70 antibodies lowered the internalization of GV1001, indicating that the interaction is critical for the efficient internalization of GV1001. Collectively, the results of this study suggest the pharmaceutical potential of GV1001, already proven safe in clinical trials, as a carrier for the delivery of macromolecular therapeutics into cells, in addition to its own anti-cancer activity.
authors.affiliation: Department of Microbiology and Immunology, Liver Research Institute, Cancer Research Institute and SNUMRC, College of Medicine, Seoul National University, Seoul, Republic of Korea.
authors.forename: Seoung-Ae
authors.initials: SA
authors.lastname: Lee
authors.forename: Bo-Ram
authors.initials: BR
authors.lastname: Kim
authors.forename: Bu-Kyung
authors.initials: BK
authors.lastname: Kim
authors.forename: Dong-Won
authors.initials: DW
authors.lastname: Kim
authors.forename: Won-Jun
authors.initials: WJ
authors.lastname: Shon
authors.forename: Na-Rae
authors.initials: NR
authors.lastname: Lee
authors.forename: Kyung-Soo
authors.initials: KS
authors.lastname: Inn
authors.forename: Bum-Joon
authors.initials: BJ
authors.lastname: Kim
mesh_terms: D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000494:pharmacology*; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism*; D004247:DNA / Q000378:metabolism; D016503:Drug Delivery Systems; D004705:Endocytosis / Q000187:drug effects; D055785:Gene Knockdown Techniques; D049452:Green Fluorescent Proteins / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000378:metabolism*; D006801:Humans; D008156:Luciferases / Q000378:metabolism; D046911:Macromolecular Substances / Q000378:metabolism*; D021962:Membrane Microdomains / Q000187:drug effects / Q000378:metabolism; D010446:Peptide Fragments / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D019098:Telomerase / Q000378:metabolism* / Q000494:pharmacology; D013997:Time Factors; D022223:Vaccines, Subunit / Q000494:pharmacology*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000494:pharmacology
chemical_list: D000906:Antibodies; D057846:Cell-Penetrating Peptides; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D010446:Peptide Fragments; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; D022223:Vaccines, Subunit; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; D004247:DNA; D008156:Luciferases; C514987:GV1001 peptide; D019098:Telomerase
keywords: Cell-penetrating peptide; Heat shock protein 70; Heat shock protein 90; Telomerase