Molecular Determinants of Vectofusin-1 and Its Derivatives for the Enhancement of Lentivirally Mediated Gene Transfer into Hematopoietic Stem/Progenitor Cells.
Fiche publication
Date publication
janvier 2016
Journal
The Journal of biological chemistry
Auteurs
Membres identifiés du Cancéropôle Est :
Pr BECHINGER Burkhard, Dr KICHLER Antoine
Tous les auteurs :
Majdoul S, Seye AK, Kichler A, Holic N, Galy A, Bechinger B, Fenard D
Lien Pubmed
Résumé
Gene delivery into hCD34+ hematopoietic stem/progenitor cells (HSPCs) using human immunodeficiency virus, type 1-derived lentiviral vectors (LVs) has several promising therapeutic applications. Numerous clinical trials are currently underway. However, the efficiency, safety, and cost of LV gene therapy could be ameliorated by enhancing target cell transduction levels and reducing the amount of LV used on the cells. Several transduction enhancers already exist, such as fibronectin fragments or cationic compounds. Recently, we discovered Vectofusin-1, a new transduction enhancer, also called LAH4-A4, a short histidine-rich amphipathic peptide derived from the LAH4 family of DNA transfection agents. Vectofusin-1 enhances the infectivity of lentiviral and γ-retroviral vectors pseudotyped with various envelope glycoproteins. In this study, we compared a family of Vectofusin-1 isomers and showed that Vectofusin-1 remains the lead peptide for HSPC transduction enhancement with LVs pseudotyped with vesicular stomatitis virus glycoproteins and also with modified gibbon ape leukemia virus glycoproteins. By comparing the capacity of numerous Vectofusin-1 variants to promote the modified gibbon ape leukemia virus glycoprotein-pseudotyped lentiviral vector infectivity of HSPCs, the lysine residues on the N-terminal extremity of Vectofusin-1, a hydrophilic angle of 140° formed by the histidine residues in the Schiffer-Edmundson helical wheel representation, hydrophobic residues consisting of leucine were all found to be essential and helped to define a minimal active sequence. The data also show that the critical determinants necessary for lentiviral transduction enhancement are partially different from those necessary for efficient antibiotic or DNA transfection activity of LAH4 derivatives. In conclusion, these results help to decipher the action mechanism of Vectofusin-1 in the context of hCD34+ cell-based gene therapy.
Mots clés
Amino Acid Sequence, Antigens, CD34, metabolism, Antimicrobial Cationic Peptides, chemistry, DNA, chemistry, Gene Transfer Techniques, Genetic Therapy, methods, Genetic Vectors, Glycoproteins, chemistry, HCT116 Cells, HEK293 Cells, HIV-1, metabolism, HeLa Cells, Hematopoietic Stem Cells, cytology, Histidine, chemistry, Humans, Lentivirus, Leukemia Virus, Gibbon Ape, Molecular Sequence Data, Peptides, chemistry, Sequence Homology, Amino Acid, Transduction, Genetic, Transfection
Référence
J. Biol. Chem.. 2016 Jan;291(5):2161-9