ZFN-Modified Stem Cells for HIV Eradication

ZFN 修饰干细胞用于根除 HIV

基本信息

批准号：
8876546
负责人：
PHILIP D GREGORY
金额：
$ 18.98万
依托单位：
FRED HUTCHINSON CANCER RESEARCH CENTER
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至 2017-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8876546
关键词：
Ablation Academia Acquired Immunodeficiency Syndrome Address Affect Alleles Allogenic Animals Autologous Bone Marrow Transplantation Back Basic Science Biological Assay Blood Bone Marrow Bone Marrow Transplantation CCR5 gene CD34 gene Cell Transplants Cells Clinic Clinical DNA DNA Repair Pathway Data Development Engineering Engraftment Enzymes Epidemiology Gene-Modified Generations Genes Genome Goals HIV HIV Infections HIV Nonprogressors HIV Seropositivity HIV-1 Healed Hematopoietic Human Immune response Immune system In Vitro Industry Infection Infusion procedures Integrase Knock-out Lead Lentivirus Vector Life Link Macaca Macaca nemestrina Mediating Messenger RNA Methodology Methods Modeling Molecular Analysis Monkeys Mus New Agents Nonhomologous DNA End Joining Patients Persons Population Procedures Protein Engineering Protocols documentation Reagent Research Resistance Resistance to infection Safety Series Site-Directed Mutagenesis Specificity Stem cells T-Lymphocyte Technology Testing Therapeutic Transplantation Virus Virus Diseases Work Zinc Fingers base clinical efficacy clinical practice clinically relevant design gene therapy genome editing healing human MCAM protein in vivo leukemia loss of function nonhuman primate novel strategies nuclease plasmid DNA progenitor programs receptor research study safety study site-specific integration small molecule stem success therapeutic development therapeutic transgene transgene expression vector zinc finger nuclease

项目摘要

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 edifing with zinc finger nucleases," the CCR5 gene can be distrupted in a targeted fashion in both primary T cells and hematopoiefic 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 populafion 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 disrupfion leads to undesired effects. At the conclusion of the proposed work, a clear translafional path will have been established to eradicate HIV1 by using autologous bone marrow transplantation combined with targeted CCR5 gene disruption.

该项目整合了学术界和行业中的小组的努力，以开发和执行初始测试旨在消除HIV-1的新代理。从广义上讲，努力的目的是解决既基础科学和翻译问题，这些问题代表了潜在治疗艾滋病与临床之间的障碍实践。一方面，小分子的流行病学证据和临床功效14年拮抗剂表明，人类CCR5基因的产物绝对需要生产力 HIV-1感染。此外，一个人的同种异体，完全髓骨骨髓移植删除CCR5的纯合子已从HIV阳性人完全消除了该病毒。在另外，使用一种称为“基因组用锌指核酸酶”的方法，CCR5基因可以是在原代T细胞和造血祖细胞/干细胞（HPSC）中以目标方式分散产生抑制病毒感染的敲除细胞和体内，因此可能使其可行设计一个人自己的HPSC，以抗HIV抗性。努力的具体目标是确定关键数字：移植的HPSC的哪一部分是CCR5敲除才能消除病毒？回答这个问题，需要在临床上相关的非人类灵长类动物模型中进行研究感染。锌指核酸酶将经过设计和优化，以淘汰辫子中的CCR5基因猕猴。将制定交付方法来驱动猕猴HPSC的淘汰赛。允许控制的实验，以解决“ CCR5基因敲除细胞的比例移植了HIV1的效率根除，“将建立程序和试剂，以通过靶向集成可选标记将允许携带5％CCR5敲除单元的HPSC的开始人口在活动物中选择以增加血液中抗HIV的细胞的比例。安全研究将是执行以确定基因脱离是否导致不希望的效应。在拟议的工作结束时，将建立一条清晰的翻译道路以消除 HIV1通过使用自体骨髓移植结合靶向CCR5基因破坏。