COMBINATORIAL USE OF CCR5 RIBOZYMES WITH ANTI HIV1 RNAS

描述：（改编自申请人摘要）HIV-1感染是 通过病毒经由高密度脂蛋白粘附到靶细胞表面而启动， 包膜糖蛋白与CD 4细胞表面的亲和相互作用 受体的 病毒进入发生后，相互作用的病毒包膜 蛋白质与细胞表面蛋白质被指定为共受体。 这 一种重要的融合反应是由两个新发现的辅助受体介导的 分子，CCR 5和融合素。 这些不同共受体的向性 位于HIV的包膜区域。 嗜T细胞病毒倾向于使用 融合跨膜蛋白，而嗜巨噬细胞病毒主要 使用趋化因子受体CCR 5。 最近，已经证明， CCR 5基因编码序列内存在32个碱基对缺失 在高加索人群中的发生率约为20%。 纯合 缺失发生在约1%的高加索人群中。 这些人 在细胞表面不表达任何CCR 5。 个人窝藏 纯合缺失对HIV-1感染具有显著抗性，但在其它方面 没有已知的健康缺陷。 对于这个等位基因是杂合子的个体， 虽然不能免疫HIV-1感染，但似乎有一个长期的过程， 发展为艾滋病和艾滋病毒相关并发症。 这些观测 导致了这里要检验的假设，即CCR 5的下调是由一种 核酶可能对预防和管理 艾滋病毒感染。 拟议的研究将检查HIV-1保护性 靶向CCR 5的核酶与抗HIV-1抗体组合的作用 核酶和体外进化的Rev结合元件诱饵（适体）。 将使用单核细胞系来评价本发明的抗HIV-1功效。 CCR 5核酶和多价核酶/适体组合。 最终 这种新的基因疗法治疗的靶细胞是CD 34+人 造血祖细胞 这些转基因细胞将被 测试它们经历多谱系分化成HIV-1的能力 抗性细胞群体。 这些研究将导致 改进了HIV-1感染的基因治疗。 这项建议是 与Ramesh Akkina博士共同提出的一套互动提案中的项目3（项目3） 1-SCID-hu小鼠模型）和Alan Knutsen博士（项目2-体外 胸腺生成）。

DESCRIPTION: (Adapted from applicant's abstract) HIV-1 infection is initiated by attachment of the virus to the target cell surface via high affinity interactions of the envelope glycoprotein to the CD4 cell surface receptor. Viral entry occurs following interaction of the viral envelope protein with cellular surface proteins designated as co-receptors. This essential fusion reaction is mediated by two newly described co-receptor molecules, CCR5 and Fusin. The tropism for these different co-receptors resides in the envelope region of HIV. T-cell tropic viruses tend to use the Fusin transmembrane protein while macrophage tropic viruses primarily use the chemokine receptor, CCR5. Recently, it has been demonstrated that a 32 base pair deletion within the coding sequence of the CCR5 gene is present at a frequency of approximately 20% in Caucasian population. Homozygous deletions occur in about 1% of the Caucasian population. These individuals do not express any CCR5 at the cell surface. Individuals harboring the homozygous deletion are markedly resistant to HIV-1 infection, but otherwise have no known health defects. Individuals heterozygous for this allele, although not immune to HIV-1 infection, appear to have a prolonged course of progression to AIDS and HIV related complications. These observations have led to the hypothesis to be tested here that down regulation of CCR5 by a ribozyme could have a marked impact on both the prevention and management of HIV infection. The proposed studies will examine the HIV-1 protective effects of ribozymes targeted to CCR5 in combination with anti-HIV-1 ribozymes and in vitro evolved Rev binding element decoys (aptamers). Monocytic cell lines will be used to evaluate the anti-HIV-1 efficacy of the CCR5 ribozyme and multivalent ribozyme/aptamer combinations. Ultimately the target cells for this novel gene therapy treatment are CD34+ human hematopoietic progenitor cells. These genetically modified cells will be tested for their ability to undergo multilineage differentiation into HIV-1 resistant cell populations. It is intended that these studies will lead to improved gene therapy treatment for HIV-1 infection. This proposal is project 3 of an interactive set of proposals with Dr. Ramesh Akkina (Project 1-SCID-hu mouse model) and Dr. Alan Knutsen (Project 2-In vitro thymopoiesis).