STEM CELL GENE THERAPY FOR AIDS USING AN ANTI-SIV ENVELOPE ANTISENSE MOLECULE

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 在造血干细胞中引入抑制基因为抗HIV-1复制的后代细胞的长寿免疫重建提供了可能性。然而，艾滋病干细胞基因疗法的成功开发可能需要在恒河猴模型中进行临床前测试。鉴于最近的研究结果表明，猕猴TRIM5α蛋白可以抑制HIV-1感染猕猴细胞，基于SIV的慢病毒载体在转导猕猴造血干细胞方面可能比基于HIV-1的载体具有明显的优势。我们评估了基于SIV的慢病毒载体(VRX859)在体外抑制病毒复制和转导恒河猴CD34淋巴祖细胞的能力。在感染同源SIV毒株后，转导VRX859载体的CD4细胞株表现出比对照细胞600多倍的病毒复制抑制。SIV分离株SIVsmE660的抑制作用较弱。用VRX859在一定的MOI范围内转导恒河猴CD34细胞，结果与HIV载体VRX494观察到的转导效率相似。然而，当我们评估恒河猴T淋巴细胞前体细胞的转导时，我们观察到基于SIV的载体的转导更有效。与对照CD4T细胞相比，来自VRX859转导的CD34细胞的CD4T细胞强烈抑制SIVmac239的复制。这种基于SIV的载体能够有效地抑制SIV的复制，再加上它对恒河猴造血祖细胞的有效转导，使其成为SIV/猕猴模型中干细胞基因治疗试验的一个有吸引力的候选者。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The introduction of inhibitory genes into hematopoietic stem cells offers the potential for long-lived immune reconstitution with progeny cells resistant to HIV-1 replication. However, successful development of stem cell gene therapy for AIDS is likely to require preclinical testing in the rhesus macaque model. In light of recent findings demonstrating that the rhesus macaque TRIM5alpha protein inhibits infection of macaque cells by HIV-1, simian immunodeficiency (SIV)-based lentiviral vectors may have distinct advantages over HIV-1-based vectors for the transduction of macaque hematopoietic stem cells. We evaluated the ability of a SIV-based lentiviral vector (VRX859) to inhibit viral replication in vitro and to transduce rhesus CD34+ lymphoid progenitor cells. Following infection with homologous SIV strains, CD4+ cell lines transduced with the VRX859 vector exhibited over 600-fold inhibition of viral replication compared to control cells. Less inhibition was observed with the divergent SIV strain SIVsmE660. Transduction of rhesus CD34+ cells with VRX859 over a range of MOIs resulted in comparable transduction efficiency as observed with the HIV vector VRX494. However, when we evaluated transduction of rhesus T lymphocyte progenitors, we observed more efficient transduction with the SIV-based vector. CD4+ T cells derived from VRX859-transduced CD34+ cells strongly inhibited SIVmac239 replication as compared to control CD4+ T cells. The ability of this SIV-based vector to mediate potent inhibition of SIV replication, coupled with its efficient transduction of rhesus hematopoietic progenitor cells, make it an attractive candidate for trials of stem cell gene therapy in the SIV/macaque model.