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: 9891736
• 负责人: HANS-PETER KIEM
• 金额: $ 99.6万
• 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/9891736
• 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 细胞彻底改变了 许多白血病的治疗，旨在实现对任何表面表达的“即插即用”靶向 人类疾病的标志。我们有兴趣优化针对持续性 HIV-1 感染的 CAR-T 疗法。 重要的是，尽管可以应用CAR-T的血液系统恶性肿瘤的范围正在迅速扩大， 一些障碍阻碍了对 HIV+ 个体的应用。首先也是最重要的是，CAR-T 功能经常被 在迁移到组织时下调或丢失，例如限制实体瘤的靶向。这也 代表了针对位于淋巴结等部位的潜伏 HIV-1 感染细胞的关键限制， 肠道和大脑。此外，越来越多的证据表明，为了让 CAR-T 识别其同源 目标，靶细胞表面可能需要抗原表达的阈值水平。的中心目标是 我们的 U19 联盟的目标是了解 CAR T 细胞在体内的命运。在我们的项目中，我们将比较病毒特异性 针对 CD20+ 白血病的 CAR 到 CAR，富含抗原与抗原稀疏的作用机制 环境，以及 CAR-T 在迁移到二级后保持有效、目标特异性功能的能力 已知含有潜伏病毒的组织部位。我们将在我们完善的非人类研究中解决这些问题 抑制 HIV-1 感染的灵长类动物 (NHP) 模型，重点开发优化的 CD4CAR 分子 由 Riley 博士在项目 3 中提出。我们首先将 CD4CAR-T 与之前表征的 NHP 版本进行比较 在 6 只未感染的动物中取得了成功的靶向白血病的 CD20 CAR 分子（“NHP CD20 CAR-T”）。接下来，我们 将重点研究 12 种先前感染过猿/人类免疫缺陷动物的 CD4CAR-T 病毒（SHIV）并通过抗逆转录病毒治疗（ART）进行抑制。我们的研究总共有 18 名未感染者或感染者， 受抑制的动物将为促进植入的机制提供前所未有的见解， 持久性和体内功能，和/或导致抗原依赖性扩增的沉默或抑制。我们有 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。使用完善的测定法来测量组织驻留记忆 T 细胞 (TRM)，以及 通过免疫组织化学和转录分析方法，我们将表征 CD4CAR-T 的功能 低和高抗原环境（即 ART 释放之前和之后），以及 ii) 对这些活动进行基准测试 与我们的阳性对照 NHP CD20 CAR-T 相比。我们的 NHP 研究目标将应用体内的黄金标准 抑制艾滋病毒感染的模型，与项目中提出的实验高度互补 1/Wherry，项目 3/Riley，项目 4/Coughlin 和 Tebas。