TY - JOUR
T1 - G2 Cell Cycle Arrest and Cyclophilin A in Lentiviral Gene Transfer
AU - Zhang, Shangming
AU - Joseph, Guiandre
AU - Pollok, Karen
AU - Berthoux, Lionel
AU - Sastry, Lakshmi
AU - Luban, Jeremy
AU - Cornetta, Kenneth
N1 - Funding Information:
We thank Dr. Xiao-Fan Wang (Department of Pharmacology and Cancer Biology, Duke University Medical Center) for the CypA expression plasmid. The Indiana University Vector Production Facility is a NIH-designated National Gene Vector Laboratory (U42RR11148) and this work was supported, in part, by a Core Centers of Excellence in Molecular Hematology grant (PHS P50DK49218) and a core laboratory supporting grant (PHSP01HL53586). K.C. is supported in part by the Indiana Genomic Initiative created through a grant from the Lilly Endowment, Inc. We are also grateful to Yi Xu, Teresa Johnson, and Jing Yao for their technical assistance.
PY - 2006/10
Y1 - 2006/10
N2 - 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.
AB - 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.
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U2 - 10.1016/j.ymthe.2006.05.022
DO - 10.1016/j.ymthe.2006.05.022
M3 - Article
C2 - 16901758
AN - SCOPUS:33748043344
VL - 14
SP - 546
EP - 554
JO - Molecular Therapy
JF - Molecular Therapy
SN - 1525-0016
IS - 4
ER -