Zhang Shangming, Joseph Guiandre, Pollok Karen, Berthoux Lionel, Sastry Lakshmi, Luban Jeremy, Cornetta Kenneth
Department of Medical and Molecular Genetics, Indiana University School of Medicine, 975 West Walnut Street, Indianapolis, IN 46202, USA.
Mol Ther. 2006 Oct;14(4):546-54. doi: 10.1016/j.ymthe.2006.05.022. Epub 2006 Aug 9.
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.
与基于嗜肝DNA病毒的载体相比，源自人类免疫缺陷病毒1型（HIV-1）的慢病毒载体转导非分裂细胞的倾向更高。我们在此报告，染料木黄酮是一种先前已知的蛋白酪氨酸激酶（PTK）抑制剂和G2细胞周期阻滞诱导剂，它以剂量依赖的方式显著增强了慢病毒转导。通过载体表达测量，在多种人类细胞系、小鼠原代淋巴细胞以及原代人类CD34(+)外周血祖细胞中均观察到转导增加。通过定量PCR评估，载体表达的增加还与载体DNA拷贝数的增加相关。染料木黄酮介导的G2细胞周期阻滞而非PTK抑制似乎是基因转移增加的主要因素。染料木黄酮还增加了亲环素A（CypA）蛋白，这是一种对高效HIV-1感染很重要的细胞蛋白。虽然我们表明CypA(-/-) Jurkat细胞对慢病毒载体的转导能力较差，但染料木黄酮确实增加了CypA缺陷细胞中的基因转移。CypA和G2细胞周期阻滞似乎是高效慢病毒基因转移的两个独立重要因素。染料木黄酮和其他G2阻滞剂的作用可能有助于提高慢病毒基因治疗的效率。
