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.

项目摘要/摘要 这项建议寻求必要的资金，以促进丹尼尔·克莱本博士的职业过渡到 独立的学术科学家。这项提案提出的最初目标是建立在基本艾滋病毒的基础上 应聘者的病毒学、免疫学和病原体-宿主相互作用专业知识，并利用这些技能 目的是在小动物模型中进行转化努力，以实现功能性艾滋病毒治疗。此外， 职业发展计划勾勒出了一条获得执行 这一提议以及向独立的过渡。最后，从成功生成的数据 这些实验的执行将产生申请R01级资金的初步数据。 人类免疫缺陷病毒(HIV)通过快速、快速地破坏宿主细胞免疫反应 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 抗逆转录病毒疗法(ART)已经大大降低了艾滋病毒感染的发病率和死亡率，但必须 无限期服用，因为艾滋病毒持续存在于长期稳定的水库中，因此对公共健康构成重大影响 负担。清除病毒库的一种方法，称为“电击并杀死”，依赖于潜伏期反转剂。 (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LRAs的临床试验收效甚微，可能是由于天然细胞的功能障碍所致。 免疫反应。基因工程方式可能会提供一种有吸引力的替代固有免疫的方法。 利用CD4胞外区重新设计嵌合抗原受体(CAR)T细胞以靶向HIV(CD4-CAR) 代表了一种被证明具有增强的细胞毒性功能的抗逃逸“杀伤”机制。因此， 我们假设，CD4-CAR T细胞将是一种有希望的干预手段，能够抑制HIV 复制，并有效地针对病毒库。 为了快速测试和优化我们的方法，我们利用了BLT人源化的HIV感染小鼠模型。 我们的初步结果表明，CD4-CAR T细胞对抗原的反应很强，扩增到1000倍 在体内，并显著保护感染HIV的BLT小鼠免受快速的CD4+T细胞损失。此外，我们还改进了 CD4-CAR T细胞的扩增潜能和细胞毒功能，并能保护CD4-CAR T细胞 免受艾滋病毒感染。然而，尽管ART能够增强病毒抑制，CAR T细胞疗法 单独使用不能持久地抑制BLT小鼠的病毒血症。我们建议从以下几个方面解决这一问题 具体目标：1)确定病毒复制能力对CAR T细胞效力的影响；2)绘制个体发育图 体内耗尽的CAR T细胞，以及3)询问先天免疫系统在调节CAR T中的作用 细胞功能。这些目标的完成将加深我们对T细胞构成因素的理解 慢性病毒感染的功能障碍，希望将这些发现转化为有效的艾滋病毒治疗方法。