G2 cell cycle arrest and cyclophilin A in lentiviral gene transfer.
Lentiviral vectors derived from the human immunodeficiency virus-1 (HIV-1) have a higher propensity to transduce nondividing cells compared to vectors based on oncoretroviruses. We report here that genistein, a previously known protein tyrosine kinase (PTK) inhibitor and G2 cell cycle arrest inducer, significantly enhanced lentiviral transduction in a dose-dependent manner. Increased transduction, as measured by vector expression, was seen in a variety of human cell lines, murine primary lymphocytes, and primary human CD34(+) peripheral blood progenitor cells as well. Increased vector expression was also associated with an increase in vector DNA copy number, as assessed by quantitative PCR. Genistein-mediated G2 cell cycle arrest, rather than PTK inhibition, appears to be the major factor responsible for increased gene transfer. Genistein also increases cyclophilin A (CypA) protein, a cellular protein important for efficient HIV-1 infection. While we show that CypA(-/-) Jurkat cells transduce poorly with lentiviral vectors, genistein does increase gene transfer in CypA-deficient cells. CypA and G2 cell cycle arrest appear to be two independent factors important for efficient lentiviral gene transfer. The role of genistein and other G2-arresting agents may be useful for improving the efficiency of lentiviral gene therapy.
Zhang
Shangming
S
Department of Medical and Molecular Genetics, Indiana University School of Medicine, 975 West Walnut Street, Indianapolis, IN 46202, USA.
Joseph
Guiandre
G
Pollok
Karen
K
Berthoux
Lionel
L
Sastry
Lakshmi
L
Luban
Jeremy
J
Cornetta
Kenneth
K
D000818:Animals; D002478:Cells, Cultured; D021984:Cyclophilin A / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D016195:G2 Phase / Q000187:drug effects; D018628:Gene Dosage; D015870:Gene Expression / Q000187:drug effects; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D019833:Genistein; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D051379:Mice; D013997:Time Factors
D019833:Genistein; D021984:Cyclophilin A