title: Comparison Between Several Integrase-defective Lentiviral Vectors Reveals Increased Integration of an HIV Vector Bearing a D167H Mutant.
abstract: HIV-1 derived vectors are among the most efficient for gene transduction in mammalian tissues. As the parent virus, they carry out vector genome insertion into the host cell chromatin. Consequently, their preferential integration in transcribed genes raises several conceptual and safety issues. To address part of these questions, HIV-derived vectors have been engineered to be nonintegrating. This was mainly achieved by mutating HIV-1 integrase at functional hotspots of the enzyme enabling the development of streamlined nuclear DNA circles functional for transgene expression. Few integrase mutant vectors have been successfully tested so far for gene transfer. They are cleared with time in mitotic cells, but stable within nondividing retina cells or neurons. Here, we compared six HIV vectors carrying different integrases, either wild type or with different mutations (D64V, D167H, Q168A, K186Q+Q214L+Q216L, and RRK262-264AAH) shown to modify integrase enzymatic activity, oligomerization, or interaction with key cellular cofactor of HIV DNA integration as LEDGF/p75 or TNPO3. We show that these mutations differently affect the transduction efficiency as well as rates and patterns of integration of HIV-derived vectors suggesting their different processing in the nucleus. Surprisingly and most interestingly, we report that an integrase carrying the D167H substitution improves vector transduction efficiency and integration in both HEK-293T and primary CD34+ cells.
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] INSERM, UMS27, Fontenay-aux-Roses, France [3] Institute of Pure and Applied Biology, Microbiology Division, Bahauddin Zakariya University Multan, Pakistan.
authors.forename: Muhammad
authors.initials: M
authors.lastname: Qamar Saeed
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] Laboratoire des maladies neurodégénératives CNRS, CEA URA 2210.
authors.forename: Noelle
authors.initials: N
authors.lastname: Dufour
authors.affiliation: Department of Translasional Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Bartholmae
authors.affiliation: CEA, Laboratoire des Cellules Souches Hématopoïétiques et Leucémiques, (IRCM), Fontenay-aux-Roses, France.
authors.forename: Urzula
authors.initials: U
authors.lastname: Sieranska
authors.affiliation: Department of Translasional Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Malaika
authors.initials: M
authors.lastname: Knopf
authors.affiliation: EBIM, Ecole Normale Supérieure de Cachan, Cachan, Paris, France.
authors.forename: Eloïse
authors.initials: E
authors.lastname: Thierry
authors.affiliation: EBIM, Ecole Normale Supérieure de Cachan, Cachan, Paris, France.
authors.forename: Sylvain
authors.initials: S
authors.lastname: Thierry
authors.affiliation: EBIM, Ecole Normale Supérieure de Cachan, Cachan, Paris, France.
authors.forename: Olivier
authors.initials: O
authors.lastname: Delelis
authors.affiliation: NewVectys, Villebon-sur-Yvette, France.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Grandchamp
authors.affiliation: NewVectys, Villebon-sur-Yvette, France.
authors.forename: Héloïse
authors.initials: H
authors.lastname: Pilet
authors.affiliation: ICM-BBL, CNRS UMR 7225, Hôpital Pitié-Salpêtrière, Paris, France.
authors.forename: Philippe
authors.initials: P
authors.lastname: Ravassard
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] INSERM, UMS27, Fontenay-aux-Roses, France.
authors.forename: Julie
authors.initials: J
authors.lastname: Massonneau
authors.affiliation: CEA, Laboratoire des Cellules Souches Hématopoïétiques et Leucémiques, (IRCM), Fontenay-aux-Roses, France.
authors.forename: Françoise
authors.initials: F
authors.lastname: Pflumio
authors.affiliation: Department of Translasional Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: von Kalle
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] INSERM, UMS27, Fontenay-aux-Roses, France.
authors.forename: François
authors.initials: F
authors.lastname: Lachapelle
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] Laboratoire des maladies neurodégénératives CNRS, CEA URA 2210.
authors.forename: Alexis-Pierre
authors.initials: AP
authors.lastname: Bemelmans
authors.affiliation: Department of Translasional Oncology, National Center for Tumor Diseases (NCT) and German Cancer Research Center (DKFZ), Heidelberg, Germany.
authors.forename: Manfred
authors.initials: M
authors.lastname: Schmidt
authors.affiliation: 1] CEA, DSV, IBM, Molecular Imaging Research Center (MIRCen), Fontenay-aux-Roses, France [2] INSERM, UMS27, Fontenay-aux-Roses, France.
authors.forename: Ché
authors.initials: C
authors.lastname: Serguera