Nongenotoxic conditioning to enhance stem cell engineering and virus-specific immunity in nonhuman primates

非基因毒性调理可增强非人灵长类动物的干细胞工程和病毒特异性免疫力

基本信息

批准号：
10601087
负责人：
HANS-PETER KIEM
金额：
$ 51.82万
依托单位：
UNIVERSITY OF SOUTHERN CALIFORNIA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-05-15 至 2025-04-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10601087
关键词：
Address Alleles Allogenic Animal Model Animals Autologous Autopsy B-Lymphocytes Bar Codes Berlin Binding Bone Marrow Boston Brain CCR5 gene CD4 Antigens Cell Transplantation Cells Cellular Immunity Clinical Clustered Regularly Interspaced Short Palindromic Repeats DNA Repair Pathway Data Development Disease remission Dose Engraftment Experimental Designs Flow Cytometry Gene Modified Genes Genetic Genetic Engineering Goals HIV HIV Infections Hematopoietic stem cells Humoral Immunities Immune Immune response Immune system Immunity Immunohistochemistry Impairment Infection Infusion procedures Kinetics Letters London Measures Methodology Methods Mississippi Modeling Mutate Nonhomologous DNA End Joining Oligonucleotides Patients Pharmaceutical Preparations Phenotype Population Progenitor Cell Engraftment Property Protocols documentation Regimen Resistance Rosaniline Dyes Sentinel Site Source Stem cell transplant T-Lymphocyte Therapeutic Time Tissues Transgenes Transplantation Viral Viral reservoir Viremia Virus Virus Activation Virus Diseases Virus Latency Whole-Body Irradiation Work antibody mimetics antiretroviral therapy cell type chimeric antigen receptor chimeric antigen receptor T cells conditioning cryogel deep sequencing delta receptors design engineered stem cells experience experimental study gene therapy hematopoietic differentiation improved in vivo inhibitor interest lymph nodes neutralizing antibody news nonhuman primate novel peripheral blood preference rational design receptor repaired resistance gene response scaffold simian human immunodeficiency virus stem cell expansion stem cells synergism therapeutic transgene trafficking

项目摘要

ABSTRACT Although hematopoietic stem and progenitor cell (HSPC) transplantation now underlies two clinical cases of HIV- 1 remission/functional cure, a means to apply this approach to a wider array of patients has not yet been identified. In this project, we will address a key limitation for HSPC-based anti-HIV strategies: the engraftment and potency of gene-edited HSPC and their progeny. Although our previous findings demonstrate that gene edited HSPCs engraft long-term in vivo, only a limited proportion persist over time, and are incapable of supporting antiretroviral therapy (ART)-free virus remission. To address this, we have i) adapted a more advanced strategy to edit our locus of interest, CCR5, ii) identified an approach to not only disrupt the CCR5 gene, but simultaneously insert therapeutic anti-HIV transgenes, and iii) designed experiments to evaluate this strategy in our robust nonhuman primate (NHP) model of suppressed HIV infection. We will target two rationally designed, highly potent anti-HIV transgenes to the gene-edited CCR5 locus: the virus-specific chimeric antigen receptor CD4CAR, and the broadly neutralizing antibody-like molecule eCD4-Ig. Our preliminary data demonstrate our ability to insert defined genetic sequences at up to 50% of targeted CCR5 alleles in primary NHP HSPCs. Here, we will optimize our approach to insert CD4CAR or eCD4-Ig, and safely engraft an autologous HSPC product containing both CD4CAR∆CCR5 and eCD4-Ig∆CCR5 HSPCs into the same animal. As we are introducing two therapeutic transgenes and simultaneously disrupting the CCR5 coreceptor, we refer to this as a “three for one” approach. In addition to generating a potent and efficiently modified HSPC product, we will work closely with each project in our U19 consortium. We will coordinate with Project 3/Cannon to identify the most efficient means to modify HSPCs, prior to in vivo studies in our respective animal models. With Project 1/Scadden, we will evaluate a bone marrow cryogel (BMC) scaffold designed to enhance the differentiation of HSPC-derived T-cells, namely CD4CAR∆CCR5 T-cells. Finally, we will investigate the impact of safer, nongenotoxic conditioning (NGC) regimens characterized by Project 2/Magenta on infection with simian/human immunodeficiency virus (SHIV) and suppression by ART. We believe that safe and efficacious engraftment of gene-modified, virus-specific HSPCs and their progeny will enable robust protection against de novo SHIV challenge, and significantly impact viral reservoirs in infected, suppressed animals.

抽象的尽管造血干细胞和祖细胞（HSPC）移植现在是两个艾滋病毒临床病例的基础 1 缓解/功能性治愈，尚未找到将该方法应用于更广泛患者的方法在这个项目中，我们将解决基于 HSPC 的抗 HIV 策略的一个关键限制：植入。以及基因编辑的 HSPC 及其后代的效力，尽管我们之前的研究结果证明了该基因。编辑后的 HSPCs 可以长期植入体内，但只有有限的一部分能够随着时间的推移而持续存在，并且无法支持无抗逆转录病毒治疗 (ART) 的病毒缓解为了解决这个问题，我们 i) 采取了更多措施。编辑我们感兴趣的位点 CCR5 的先进策略，ii) 确定了一种不仅破坏 CCR5 的方法基因，但同时插入治疗性抗 HIV 转基因，并且 iii) 设计实验来评估这一点在我们抑制 HIV 感染的稳健非人类灵长类动物 (NHP) 模型中，我们将合理地针对两个目标。针对基因编辑的 CCR5 位点设计的高效抗 HIV 转基因：病毒特异性嵌合抗原受体 CD4CAR 和广泛中和抗体样分子 eCD4-Ig 我们的初步数据。证明我们能够在原代细胞中插入高达 50% 的目标 CCR5 等位基因的确定基因序列在这里，我们将优化插入 CD4CAR 或 eCD4-Ig 的方法，并安全地植入将含有 CD4CARΔCCR5 和 eCD4-IgΔCCR5 HSPC 的自体 HSPC 产品注入同一动物体内。当我们引入两种治疗性转基因并同时破坏 CCR5 辅助受体时，我们参考将此视为“三合一”方法除了生成有效且高效的改性 HSPC 产品外，我们将与 U19 联盟中的每个项目密切合作，我们将与 Project 3/Cannon 协调以确定。在我们各自的动物模型中进行体内研究之前，这是修改 HSPC 的最有效方法。 1/Scadden，我们将评估一种骨髓冷冻凝胶 (BMC) 支架，旨在增强骨髓的分化 HSPC 衍生的 T 细胞，即 CD4CARΔCCR5 T 细胞最后，我们将研究更安全的影响。 Project 2/Magenta 描述的针对猿类/人类感染的非基因毒性调理 (NGC) 方案我们相信，免疫缺陷病毒（SHIV）的安全和有效的植入以及通过 ART 进行抑制。基因修饰的病毒特异性 HSPC 及其后代将能够针对新发 SHIV 提供强有力的保护挑战，并显着影响受感染、抑制动物的病毒库。