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的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 将抑制基因引入造血干细胞中为后代细胞抵抗HIV-1复制提供了长期免疫重建的可能性。 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试验。 鉴于最近的发现表明恒河猴TRIM 5 α蛋白抑制猕猴细胞被HIV-1感染，基于猿免疫缺陷（SIV）的慢病毒载体可能比基于HIV-1的载体在猕猴造血干细胞的转导中具有明显的优势。 我们评估了SIV为基础的慢病毒载体（VRX 859）在体外抑制病毒复制的能力，以及对恒河猴CD 34+淋巴祖细胞的抑制能力。在用同源SIV毒株感染后，与对照细胞相比，用VRX 859载体转导的CD 4+细胞系表现出超过600倍的病毒复制抑制。 用趋异SIV毒株SIVsmE 660观察到较少的抑制。用VRX 859在一定MOI范围内转导恒河猴CD 34+细胞导致与用HIV载体VRX 494观察到的相当的转导效率。 然而，当我们评估恒河猴T淋巴细胞祖细胞的转导时，我们观察到用基于SIV的载体进行更有效的转导。 与对照CD 4 + T细胞相比，源自VRX 859转导的CD 34+细胞的CD 4 + T细胞强烈抑制SIVmac 239复制。 这种基于SIV的载体介导有效抑制SIV复制的能力，加上其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.