ZFN-Modified Stem Cells for HIV Eradication

该项目整合了学术界和工业界团体的努力，以开发和执行初始测试 一种旨在根除HIV-1的新药剂。从广义上讲，这一努力的目标是解决这两个问题， 基础科学和转化问题代表了潜在的艾滋病治愈方法和临床治疗方法之间的障碍， 实践一方面，14年的流行病学证据和临床疗效的小分子 拮抗剂已经表明，人CCR 5基因的产物是生产性细胞所绝对需要的。 感染HIV-1。此外，同种异体，完全清髓性骨髓移植从一个人， CCR 5缺失的纯合子已经完全根除了HIV阳性者的病毒。在 此外，使用一种称为“锌指核酸酶基因组编辑”的方法，可以将CCR 5基因 在原代T细胞和造血祖细胞/干细胞（HPSC）中以靶向方式分散， 产生离体和体内抗病毒感染的敲除细胞，从而潜在地使其可行， 设计一个人自己的HPSC以抵抗艾滋病毒。这项工作的具体目标是确定一个关键的 数量：移植的HPSC需要CCR 5敲除以根除病毒的比例是多少？回答 对于这个问题，需要在临床相关的非人灵长类HIV模型中进行研究 感染将对锌指核酸酶进行工程化和优化，以敲除尾纤中的CCR 5基因。 猕猴。将制定递送方法以在猕猴HPSC中驱动该敲除。以允许 对照实验，以解决“移植的CCR 5敲除细胞的分数与HIV 1的效率 根除，”程序和试剂将建立敲除CCR 5通过有针对性的整合， 选择性标记，其将允许携带5%CCR5敲除细胞的HPSC的起始群体被 在活体动物中选择，以增加血液中抗艾滋病毒细胞的比例。安全性研究将 以确定基因破坏是否导致不期望的效果。 在拟议的工作结束时，将建立一个明确的转化路径，以消除 采用自体骨髓移植联合靶向CCR 5基因破坏治疗HIV 1。

The project integrates efforts from groups in academia and in industry to develop and perform initial testing of a new agent aimed at the eradication of HIV-1. In broad terms, the objective of the effort is to resolve both basic science and translational issues that represent obstacles between a potential cure for AIDS and clinical practice. On the one hand, 14 years of epidemiologic evidence and clinical efficacy of a small-molecule antagonist have shown that the product of the human CCR5 gene is absolutely required for productive infection by HIV-1. Furthermore, an allogeneic, fully myeloablative bone marrow transplant from a person homozygous for a deletion of CCR5 has completely eradicated the virus from an HIV-positive person. In addition, using an approach known as "genome editing with zinc finger nucleases," the CCR5 gene can be distrupted in a targeted fashion in both primary T cells and hematopoietic progenitor/stem cells (HPSCs) to produce knockout cells that resist virus infection ex vivo and in vivo, thus potentially making it feasible to engineer a person's own HPSCs for HIV resistance. The specific goal of the effort is to determine a critical number: what fraction of transplanted HPSCs need to be CCR5 knockouts to eradicate the virus? To answer this question, a study needs to be performed in a clinically relevant nonhuman primate model of HIV infection. Zinc finger nucleases will be engineered and optimized to knock out the CCR5 gene in the pigtailed macaque. Delivery methodology will be worked out to drive this knockout in macaque HPSCs. To allow a controlled experiment in addressing the "fraction of CCR5 knockout cells transplanted vs efficiency of HIV1 eradication," procedures and reagents will be built to knock out CCR5 via the targeted integration of a selectable marker that will allow a starting population of HPSCs harboring 5% CCR5-knockout cells to be selected for in live animals to increase the fraction of HIV-resistant cells in the blood. Safety studies will be performed to determine whether the gene disruption leads to undesired effects. At the conclusion of the proposed work, a clear translational path will have been established to eradicate HIV1 by using autologous bone marrow transplantation combined with targeted CCR5 gene disruption.