Modeling HIV CAR-T cell trafficking and persistence in Non-Human Primates

模拟 HIV CAR-T 细胞在非人类灵长类动物中的运输和持久性

• 批准号: 10165495
• 负责人: HANS-PETER KIEM
• 金额: $ 96.84万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 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/10165495
• 关键词: Address; Animals; Antigens; Autologous; Autopsy; B-Cell Leukemia; B-Lymphocytes; Benchmarking; Biological Assay; Brain; CCR5 gene; CD19 gene; CXCR4 gene; Cell Line; Cell Therapy; Cell surface; Cells; Chimeric Proteins; Chromosome Mapping; Cryopreservation; Data; Engraftment; Environment; Flow Cytometry; Gene Expression Profiling; Gene-Modified; Genes; Goals; Gold; Gut associated lymphoid tissue; HIV; HIV Infections; Hematologic Neoplasms; Hematopoietic stem cells; Immune; Immunohistochemistry; Individual; Infection; Injections; Lead; MS4A1 gene; Macaca; Measurement; Measures; Mediating; Modeling; Modification; Neuraxis; Nucleic Acids; Paraffin Embedding; Patients; Pattern; Phenotype; Play; Property; Research; Resistance to infection; Sampling; Secondary to; Signal Transduction; Site; Solid Neoplasm; Surface; Surface Antigens; T memory cell; T-Lymphocyte; Tissues; Tumor Antigens; Viral reservoir; Virus; Virus Diseases; Virus Latency; Virus Replication; Withdrawal; Work; antiretroviral therapy; base; chimeric antigen receptor; chimeric antigen receptor T cells; density; design; engineered T cells; experimental study; gene therapy; human disease; in vivo; in vivo Model; insight; interest; leukemia; lymph nodes; migration; nonhuman primate; paraform; prevent; simian human immunodeficiency virus; success; trafficking; tumor microenvironment; zinc finger nuclease

Project 2 - Abstract Modification of autologous T-cells with chimeric antigen receptor (CAR) molecules has revolutionized the treatment of many leukemias, and is designed to enable “plug and play” targeting of any surface-expressed marker of human disease. We are interested in optimizing CAR-T therapies for persistent HIV-1 infection. Importantly, although the list of hematological malignancies to which CAR-T can be applied is rapidly expanding, several barriers have prevented application to HIV+ individuals. First and foremost, CAR-T function is frequently downregulated or lost upon migration to tissues, for example limiting targeting of solid tumors. This also represents a key limitation for targeting of latently HIV-1 infected cells that reside at sites including lymph nodes, gut, and the brain. Furthermore, increasing evidence suggests that in order for CAR-T to recognize their cognate targets, a threshold level of antigen expression may be required at the target cell surface. The central goal of our U19 consortium is to understand the fate of a CAR T-cell in vivo. In our project, we will compare virus-specific CARs to CARs directed against CD20+ leukemias, mechanisms of action in antigen-rich vs. antigen-sparse environments, and the ability of CAR-T to maintain potent, target-specific function after migrating to secondary tissue sites known to harbor latent virus. We will address these questions in our well-established nonhuman primate (NHP) model of suppressed HIV-1 infection, focusing on the optimized CD4CAR molecule developed by Dr. Riley in Project 3. We will first compare CD4CAR-T to a previously-characterized NHP version of the successful, leukemia-targeting CD20 CAR molecule (“NHP CD20 CAR-T”), in 6 uninfected animals. Next, we will focus on CD4CAR-T in 12 animals that have been previously infected with simian/human immunodeficiency virus (SHIV) and suppressed by antiretroviral therapy (ART). Our studies in a total of 18 uninfected or infected, suppressed animals will provide unprecedented insights into the mechanisms that promote engraftment, persistence, and function in vivo, and/or lead to silencing or inhibition of antigen-dependent expansion. We have chosen the NHP model for our studies, as a key aspect of our approach is to better understand CAR-T trafficking and function in tissues. Using well-established assays to measure tissue resident memory T-cells (TRM), along with immunohistochemistry and transcriptional profiling approaches, we will characterize CD4CAR-T function in low and high antigen environments (i.e. before and after release of ART), and ii) benchmark these activities against our positive control, NHP CD20 CAR-T. Our NHP research aims will apply the gold standard for in vivo modeling of suppressed HIV infection, and are highly complementary to experiments proposed in Project 1/Wherry, Project 3/Riley, and Project 4/Coughlin and Tebas.

项目2 -摘要 用嵌合抗原受体（CAR）分子修饰自体T细胞已经彻底改变了免疫学。 治疗许多白血病，并被设计为能够“即插即用”靶向任何表面表达的 人类疾病的标志。我们有兴趣优化CAR-T治疗持续性HIV-1感染。 重要的是，尽管CAR-T可以应用于的血液恶性肿瘤的名单正在迅速扩大， 有几个障碍阻碍了对HIV阳性个体的应用。首先，CAR-T功能经常被 在迁移至组织时下调或丢失，例如限制实体瘤的靶向。这也 代表了靶向潜伏HIV-1感染的细胞的关键限制，所述潜伏HIV-1感染的细胞位于包括淋巴结的部位， 内脏和大脑此外，越来越多的证据表明，为了使CAR-T识别其同源物， 在靶细胞中，在靶细胞表面可能需要阈值水平的抗原表达。的中心目标 我们的U19联盟是为了了解体内CAR T细胞的命运。在我们的项目中，我们将比较病毒特异性 针对CD 20+白血病的汽车到汽车，富含抗原与缺乏抗原的作用机制 环境，以及CAR-T在迁移到第二免疫系统后维持有效的靶特异性功能的能力。 潜伏病毒的组织部位我们将解决这些问题，在我们完善的非人类 抑制HIV-1感染的灵长类动物（NHP）模型，重点是开发的优化的CD 4CAR分子 作者：项目3中的莱利博士。我们将首先将CD 4CAR-T与先前表征的NHP版本进行比较。 在6只未感染的动物中成功地靶向白血病的CD 20 CAR分子（“NHP CD 20 CAR-T”）。接下来我们 将重点研究12只先前感染过猿猴/人类免疫缺陷病毒的动物的CD 4CAR-T。 病毒（SHIV）和抑制抗逆转录病毒疗法（ART）。我们的研究共涉及18名未感染或感染者， 抑制的动物将为促进移植的机制提供前所未有的见解， 持久性和体内功能，和/或导致沉默或抑制抗原依赖性扩增。我们有 我选择NHP模型进行研究，因为我们方法的一个关键方面是更好地了解CAR-T贩运 并在组织中发挥作用。使用完善的测定来测量组织驻留记忆T细胞（TRM），沿着 通过免疫组织化学和转录谱分析方法，我们将表征CD 4CAR-T在 低和高抗原环境（即ART释放之前和之后），和ii）基准这些活动 针对我们的阳性对照NHP CD 20 CAR-T。我们的NHP研究目标将适用于体内的黄金标准 抑制艾滋病毒感染的建模，并高度补充项目中提出的实验 1/Wherry，Project 3/Riley，and Project 4/Coughlin and Tebas.