Akkina Ramesh, Banerjea Akhil, Bai Jirong, Anderson Joseph, Li Ming-Jie, Rossi John
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, CO 80523-1619, USA.
Anticancer Res. 2003 May-Jun;23(3A):1997-2005.
Gene therapy strategies for HIV infection require gene transduction of hematopoietic stem cells with effective therapeutic constructs. Here we summarize our studies on anti-HIV ribozymes, RNA decoys and the newly described siRNAs. The therapeutic constructs consisted of an anti-CCR5 ribozyme to down-regulate the HIV-1 cell surface co-receptor and ribozymes targeted to viral mRNAs coding for the tat, rev and env proteins. The RNA decoy targeted rev and the siRNA was directed against a sequence common to rev and tat mRNAs. CD34 hematopoietic progenitor cells were transduced with retroviral or lentiviral vectors containing these constructs. They were differentiated into macrophages in vitro and T cells in vivo in a SCID-hu mouse thymopoiesis model. The transgene-containing macrophages and T cells were found to be phenotypically normal. When challenged in vitro with HIV-1, they showed significant anti-viral resistance. These proof-of-concept studies demonstrated the utility of RNA-based anti-HIV constructs for gene therapy.
针对HIV感染的基因治疗策略需要用有效的治疗性构建体对造血干细胞进行基因转导。在此，我们总结了我们关于抗HIV核酶、RNA诱饵和新描述的小干扰RNA（siRNA）的研究。治疗性构建体由一种抗CCR5核酶组成，用于下调HIV-1细胞表面共受体，以及靶向编码tat、rev和env蛋白的病毒mRNA的核酶。RNA诱饵靶向rev，而siRNA则针对rev和tat mRNA共有的序列。用含有这些构建体的逆转录病毒或慢病毒载体转导CD34造血祖细胞。在SCID-hu小鼠胸腺生成模型中，它们在体外分化为巨噬细胞，在体内分化为T细胞。发现含有转基因的巨噬细胞和T细胞在表型上是正常的。当在体外受到HIV-1攻击时，它们表现出显著的抗病毒抗性。这些概念验证研究证明了基于RNA的抗HIV构建体在基因治疗中的实用性。
