Overcoming the hurdles to successful CAR T cell therapy for a functional HIV cure

克服成功 CAR T 细胞疗法实现功能性 HIV 治愈的障碍

• 批准号: 10013942
• 负责人: Daniel Thomas Claiborne
• 金额: $ 16.2万
• 依托单位: WISTAR INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10013942
• 关键词: Address; Affect; Animal Model; Anti-Retroviral Agents; Antigens; Automobile Driving; BLT mice; Biological Models; Blood Cells; CAR T cell therapy; CD4 Positive T Lymphocytes; Career Mobility; Cell Therapy; Cell physiology; Cells; Cellular biology; Chronic; Clinic; Clinical Trials; Complex; Cytotoxic T-Lymphocytes; Data; Development; Development Plans; Disease; Engineering; Functional disorder; Funding; Generations; Genetic Engineering; Goals; Growth; HIV; HIV Infections; HIV therapy; Human; Immune; Immune System Diseases; Immune response; Immune system; Immunity; Immunology; Individual; Innate Immune System; Intervention; Knowledge; Malignant Neoplasms; Maps; Methods; Modality; Modeling; Morbidity - disease rate; Natural Immunity; Nature; Pattern; Pharmaceutical Preparations; Phenotype; Play; Preventive vaccine; Public Health; Resistance; Role; Science; Scientist; Shock; Source; T cell response; T memory cell; T-Cell Activation; T-Lymphocyte; Testing; Therapeutic; Time; Translating; Variant; Viral; Viral Load result; Viral reservoir; Viremia; Virus; Virus Diseases; Virus Replication; adaptive immunity; antigen-specific T cells; antiretroviral therapy; cancer cell; career; career development; chimeric antigen receptor; chimeric antigen receptor T cells; cytotoxic; exhaust; exhaustion; experimental study; humanized mouse; immune activation; immune reconstitution; immunopathology; improved; in vivo; invention; monocyte; mortality; mouse model; novel; pathogen; programmed cell death protein 1; purge; receptor; skills; success; transcription factor; transcriptome sequencing; transcriptomics; viral fitness; virology

Project Summary/Abstract This proposal seeks the necessary funding to facilitate the career transition of Dr. Daniel Claiborne to an independent academic scientist. The original aims brought forth in this proposal build upon the basic HIV virology, immunology, and pathogen-host interaction expertise of the candidate and leverage these skills towards translational efforts in a small animal model aimed at achieving a functional HIV cure. Furthermore, the career development plan outlines a path towards acquiring valuable skills necessary for the execution of this proposal as well as the transition to independence. Finally, the data generated from the successful execution of these experiments will yield the preliminary data to apply for R01-level funding. The human immunodeficiency virus (HIV) promptly subverts the host cellular immune response through rapid viral escape and high antigen loads leading to chronic immune activation and T cell dysfunction. Combination antiretroviral therapy (ART) has drastically reduced the morbidity and mortality of HIV infection, but must be taken indefinitely as HIV persists in long-lived stable reservoirs and thus presents a significant public health burden. One approach to purging the viral reservoir, termed “shock and kill”, relies on latency reversing agents (LRAs) to reactivate latently infected cells such that they can be cleared by the immune system. However, clinical trials testing LRAs have achieved little success, perhaps due to the dysfunction of the natural cellular immune response. Genetic engineering modalities may offer an attractive alternative to intrinsic immunity. Chimeric antigen receptor (CAR) T cells re-engineered to target HIV using the CD4 ectodomain (CD4-CAR) represent an escape-resistant “kill” mechanism demonstrated to have enhanced cytotoxic function. Therefore, we hypothesized that CD4-CAR T cells would be a promising intervention capable of suppressing HIV replication and effectively targeting the viral reservoir. To rapidly test and optimize our approach, we made use of the BLT humanized mouse model of HIV infection. Our preliminary results demonstrate that CD4-CAR T cells respond robustly to antigen, expand up to 1000-fold in vivo, and significantly protect HIV-infected BLT mice from rapid CD4+ T cell loss. Furthermore, we improved the expansion potential and cytotoxic function of CD4-CAR T cells, and were able to protect CD4-CAR T cells from HIV infection. However, despite their ability to enhance viral suppression with ART, CAR T cell therapy alone was unable to durably suppress viremia in BLT mice. We propose to address this deficit in the following specific aims: 1) Determine the effect of viral replication capacity on CAR T cell efficacy, 2) map the ontogeny of exhausted CAR T cells in vivo, and 3) interrogate the role of the innate immune system in modulating CAR T cell function. Completion of these aims will enhance our understanding of the factors contributing to T cell dysfunction in chronic viral infections, with the hope of translating these findings towards a functional HIV cure.

项目概要/摘要 该提案寻求必要的资金，以促进 Daniel Claiborne 博士的职业转型。 独立学术科学家。该提案提出的最初目标建立在基本的艾滋病毒基础上 候选人的病毒学、免疫学和病原体-宿主相互作用专业知识，并利用这些技能 致力于小动物模型的转化努力，旨在实现功能性艾滋病毒治愈。此外， 职业发展计划概述了获得执行任务所需的宝贵技能的途径 这项提议以及向独立的过渡。最后成功生成的数据 这些实验的执行将产生申请 R01 级资助的初步数据。 人类免疫缺陷病毒（HIV）通过快速的免疫反应迅速破坏宿主的细胞免疫反应。 病毒逃逸和高抗原负荷导致慢性免疫激活和 T 细胞功能障碍。组合 抗逆转录病毒治疗（ART）已大大降低了艾滋病毒感染的发病率和死亡率，但必须 无限期地采取，因为艾滋病毒持续存在于长期稳定的储存库中，从而对公共卫生产生重大影响 负担。一种清除病毒库的方法称为“休克和杀死”，依赖于潜伏期逆转剂 （LRA）重新激活潜伏感染的细胞，以便它们可以被免疫系统清除。然而， 测试 LRA 的临床试验收效甚微，这可能是由于天然细胞功能障碍所致。 免疫反应。基因工程方式可能为内在免疫提供有吸引力的替代方案。 使用 CD4 胞外域 (CD4-CAR) 重新设计嵌合抗原受体 (CAR) T 细胞以靶向 HIV 代表了一种抗逃逸“杀伤”机制，被证明具有增强的细胞毒功能。所以， 我们假设 CD4-CAR T 细胞将是一种有前景的干预措施，能够抑制 HIV 复制并有效地靶向病毒库。 为了快速测试和优化我们的方法，我们利用了 HIV 感染的 BLT 人源化小鼠模型。 我们的初步结果表明 CD4-CAR T 细胞对抗原有强烈反应，扩增高达 1000 倍 体内，并显着保护 HIV 感染的 BLT 小鼠免受 CD4+ T 细胞快速损失。此外，我们还改进了 CD4-CAR T细胞的扩增潜力和细胞毒功能，并能够保护CD4-CAR T细胞 来自艾滋病毒感染。然而，尽管 CAR T 细胞疗法能够通过 ART 增强病毒抑制， 单独使用不能持久抑制 BLT 小鼠的病毒血症。我们建议通过以下方式解决这一缺陷 具体目标：1) 确定病毒复制能力对 CAR T 细胞功效的影响，2) 绘制个体发育图 体内耗尽的 CAR T 细胞，3) 探究先天免疫系统在调节 CAR T 中的作用 细胞功能。完成这些目标将增强我们对 T 细胞影响因素的理解 慢性病毒感染的功能障碍，希望将这些发现转化为功能性艾滋病毒治疗。