title: Design and characterization of novel cell-penetrating peptides from pituitary adenylate cyclase-activating polypeptide.
abstract: The discovery of cell-penetrating peptide opened up new promising avenues for the non-invasive delivery of non-permeable biomolecules within the intracellular compartment. However, some setbacks such as possible toxic effects or unexpected immunological responses have limited their use in clinic. To overcome these obstacles, we investigated the use of novel cell-penetrating peptides (CPPs) derived from the endogenous neuropeptide Pituitary adenylate cyclase-activating polypeptide (PACAP). First, we demonstrated the propensity of native PACAP isoforms (PACAP27 and PACAP38) to efficiently deliver a large and non-permeable molecule, i.e. streptavidin, into cells. An inactive modified fragment of PACAP38, i.e. [Arg(17)]PACAP(11-38), with preserved cell-penetrating physico-chemical properties, was also synthesized and successfully use for the intracellular delivery of various cargoes such as small molecules, peptides, proteins, and polynucleotides. Especially, its effectiveness as a transfection agent was comparable to Lipofectamine 2000 while being non-toxic for cells. Uptake mechanism studies demonstrated that direct translocation, caveolae-dependent endocytosis and macropinocytosis were involved in the internalization of [Arg(17)]PACAP(11-38). This study not only opened up a new aspect in the usefulness of PACAP and its derivatives for therapeutic application but also contributed to the identification of new members of the CPP family. As such, inactive PACAP-related analogs could represent excellent vectors for in vitro and in vivo applications.
authors.affiliation: INRS-Institut Armand-Frappier, Université du Québec, 531 boulevard des Prairies, Ville de Laval, Québec, Canada H7V 1B7.
authors.forename: Ngoc-Duc
authors.initials: ND
authors.lastname: Doan
authors.forename: Myriam
authors.initials: M
authors.lastname: Létourneau
authors.forename: David
authors.initials: D
authors.lastname: Vaudry
authors.forename: Nicolas
authors.initials: N
authors.lastname: Doucet
authors.forename: Benjamin
authors.initials: B
authors.lastname: Folch
authors.forename: Hubert
authors.initials: H
authors.lastname: Vaudry
authors.forename: Alain
authors.initials: A
authors.lastname: Fournier
authors.forename: David
authors.initials: D
authors.lastname: Chatenet
mesh_terms: D000818:Animals; D016466:CHO Cells; D057846:Cell-Penetrating Peptides / Q000008:administration & dosage*; D006224:Cricetinae; D003412:Cricetulus; D015195:Drug Design; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide / Q000737:chemistry*; D010957:Plasmids; D014162:Transfection / Q000379:methods*
chemical_list: D057846:Cell-Penetrating Peptides; D051219:Pituitary Adenylate Cyclase-Activating Polypeptide; D049452:Green Fluorescent Proteins