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，以开发和执行初步测试 一种旨在根除艾滋病毒-1的新制剂的研发。概括地说，这一努力的目标是解决这两个问题 基础科学和翻译问题代表了艾滋病的潜在治疗方法和临床之间的障碍 练习一下。一方面，14年的流行病学证据和小分子药物的临床疗效 拮抗剂已经证明，人类CCR5基因的产物是生产所必需的 感染HIV-1病毒。此外，完全清髓的同种异体骨髓移植 CCR5缺失的纯合子已经从HIV阳性者身上完全根除了病毒。在……里面 此外，使用一种被称为“锌指核酸酶基因组编辑”的方法，CCR5基因可以 在原代T细胞和造血祖/干细胞(HPSCs)中定向分离以 产生能够抵抗体内和体外病毒感染的基因敲除细胞，从而有可能实现 设计一个人自己的hPSCs来抵抗艾滋病毒。这项工作的具体目标是确定一个关键的 数字：移植的hPSCs中需要有多大比例的CCR5基因敲除才能根除病毒？回答 这个问题，需要在临床上相关的非人类灵长类艾滋病毒模型中进行研究 感染。锌指核酸酶将被改造和优化，以敲除尾辫中的CCR5基因 猕猴。将制定出在猕猴hPSC中进行这种敲除的递送方法。要允许一个 CCR5基因敲除细胞移植比例与HIV1效率的对照实验 将建立程序和试剂，通过有针对性的整合 可选择的标记，将允许携带5%CCR5基因敲除细胞的hPSC的起始群体 在活体动物中选择，以增加血液中抗艾滋病毒细胞的比例。安全研究将会是 以确定基因中断是否会导致不想要的效果。 在拟议的工作结束时，将建立一条明确的翻译路径来根除 应用自体骨髓移植联合靶向CCR5基因阻断治疗HIV1。

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.