Comparison Between Several Integrase-defective Lentiviral Vectors Reveals Increased Integration of an HIV Vector Bearing a D167H Mutant

• Main Authors: Muhammad Qamar Saeed (Author), Noelle Dufour (Author), Cynthia Bartholmae (Author), Urzula Sieranska (Author), Malaika Knopf (Author), Eloïse Thierry (Author), Sylvain Thierry (Author), Olivier Delelis (Author), Nicolas Grandchamp (Author), Héloïse Pilet (Author), Philippe Ravassard (Author), Julie Massonneau (Author), Françoise Pflumio (Author), Christof von Kalle (Author), François Lachapelle (Author), Alexis-Pierre Bemelmans (Author), Manfred Schmidt (Author), Ché Serguera (Author)
• Format: Book
• Published: Elsevier, 2014-01-01T00:00:00Z.
• Subjects: article
• Online Access: Connect to this object online.
• Notes: 2162-2531; 10.1038/mtna.2014.65
• Keywords: CD34+ progenitors; integrase; integration sites; lentiviral vectors; residual integration; transduction; Therapeutics. Pharmacology; RM1-950
• Source: Molecular Therapy: Nucleic Acids, Vol 3, Iss C (2014)

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.