Modifying CMV-specificT cells with a novel bicistronic CD4-CAR/maC46 vector to target HIV

用新型双顺反子 CD4-CAR/maC46 载体修饰 CMV 特异性 T 细胞以靶向 HIV

• 批准号: 10188414
• 负责人: Nathan Michel Johnson
• 金额: $ 5.1万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-05 至 2022-07-04
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10188414
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Address; Adoptive Transfer; Affect; Animal Model; Animals; Antigens; Binding; CD4 Positive T Lymphocytes; Cell Line; Cells; Cessation of life; Chimeric Proteins; Clinic; Clinical; Clinical Trials; Cytomegalovirus; Cytomegalovirus Vaccines; Data; Disease; Down-Regulation; Effector Cell; Epidemic; Extracellular Domain; Failure; Financial Hardship; Frequencies; Goals; HIV; HIV Infections; HIV resistance; HIV-1; Heterosexuals; Human; Immune system; Immunocompromised Host; Immunodeficient Mouse; Immunologic Surveillance; Infection; Lead; Life; Life Cycle Stages; Link; MHC Class I Genes; Macaca mulatta; Membrane; Modeling; Mus; Mutation; Opportunistic Infections; Pathogenesis; Pathogenicity; Patients; Peripheral; Peripheral Blood Mononuclear Cell; Pharmacotherapy; Plasma; Population; Prevention strategy; Residual state; Retroviral Vector; Rhesus; SIV; Signal Transduction; Specificity; T-Cell Depletion; T-Lymphocyte; Therapeutic; Therapy Evaluation; Tissues; Translating; Vaccinated; Viral; Viral Load result; Viral Physiology; Viral reservoir; Viremia; Virus; Virus Replication; Work; antigen binding; antiretroviral therapy; base; cellular transduction; chimeric antigen receptor; chimeric antigen receptor T cells; clinical efficacy; cohort; cost; design; experimental study; extracellular; follow-up; gene therapy; humanized mouse; immune activation; immune function; in vivo; in vivo Model; inhibitor/antagonist; innovation; non-compliance; novel; novel therapeutics; prevent; simian human immunodeficiency virus; socioeconomics; terminally differentiated effector memory (TEM) T cells; transmission process; vector; vector vaccine; vector-based vaccine; viral fitness; viral rebound; viral transmission

Project Summary Human Immunodeficiency Virus-1 (HIV-1) has killed over 35 million people to date and infects 2 million new people each year. Infection with HIV causes depletion of CD4+ T Cells leading to an immunocompromised state, and high plasma viral load is correlated with high transmissibility. Antiretroviral therapy (ART), although effective in controlling plasma viremia and transmission, does not purge the latent or persistent reservoirs necessary to eliminate infection. Given that non-compliance, socio-economic barriers, and loss to follow up are common barriers to successful ART therapy, it is imperative to discover therapeutics that provide both lifetime suppression of viral loads and depletion of viral reservoirs. Recently, studies have demonstrated control of viral replication and decreasing viral reservoirs in 50% of rhesus vaccinated with a CMV vaccine vector. They propose that the continuous immunosurveillance of SIV by TEM cells is maintained by the persistent CMV vectors. To mimic the immunosurveillance and increase the HIV-specific CTL activity in vivo, we will genetically modify CMV-specific T cells with a chimeric antigen receptor (CAR) and follow the effects on viral reservoirs in humanized mice and rhesus macaque. The CARs express the CD4 extracellular domain to redirect CTL activity against HIV, and intracellular T cell signaling domains to stimulate CTL functions. These genetically modified T cells target a critical step in the viral life cycle independent of MHC presentation, targeting heterogeneous viruses while avoiding the potential for viral escape. Additionally, based our preliminary studies, we will include the potent fusion inhibitor maC46 to protect these genetically modified T cells from infection. In contrast to prior studies, maC46 will be incorporated bicistronically, as opposed to co-transduced. This will have the added benefit of higher percentages of transduced cells carrying both CD4-CAR and maC46. Such an innovation would result in greater ability to generate genetically modified T cells when adapted to a clinical setting. We hypothesize that CMV-specific T cells, when transduced with bicistronic CD4-CAR and fusion inhibitor, will persist in vivo based on their CMV specificity, but will be protected from infection and will target residual/reactivated HIV+ cells. These experiments would be the first use of a bicistronic vector incorporating CD4-CAR with maC46 transduced into CMV-specific T cells, and show rescue of both a humanized mouse, and rhesus macaque model. We will address this hypothesis through the following specific aims: 1) Transduce CMV-specific T cells with bicistronic CD4-CAR/maC46 and assess inhibition of viral replication, 2) Adoptively transfer CMV-specific CD4-CAR/maC46 transduced T cells in vivo using NSG mice and challenge with HIV infection, and 3) adoptively transfer CMV-specific CD4-CAR/maC46 transduced T cells into SHIV infected rhesus macaque and compare to the clinical efficacy of ART therapy in terms of plasma viral load, transmissibility, and latent reservoirs. Since rhesus macaque is an important animal model for both HIV pathogenesis and gene therapy, the evaluation of genetically modified T cells in rhesus/SHIV model will translate quickly into the clinic. The ability of genetically modified T cells to control viremia in the absence of ART, especially in the rhesus/challenge model, would be a significant advancement in HIV treatment and would strongly promote a new clinical trial for genetically modified T cells in HIV/AIDS.

项目概要 人类免疫缺陷病毒-1 (HIV-1) 迄今为止已导致超过 3500 万人死亡，并感染了 200 万人 每年的人。 HIV 感染会导致 CD4+ T 细胞耗竭，从而导致免疫功能低下 状态，高血浆病毒载量与高传染性相关。抗逆转录病毒治疗（ART），尽管 有效控制血浆病毒血症和传播，不会清除潜在或持久的病毒库 消除感染所必需的。鉴于不遵守规定、社会经济障碍和后续行动缺失等问题 成功 ART 治疗的常见障碍，必须发现能够提供终生治疗的治疗方法 抑制病毒载量和耗尽病毒库。最近，研究表明可以控制病毒 50% 接种 CMV 疫苗载体的恒河猴体内复制并减少病毒库。他们 提出 TEM 细胞对 SIV 的持续免疫监视是由持续的 CMV 维持的 向量。为了模拟免疫监视并增加体内 HIV 特异性 CTL 活性，我们将在基因上 用嵌合抗原受体 (CAR) 修饰 CMV 特异性 T 细胞，并跟踪对病毒储存库的影响 拟人化小鼠和恒河猴。 CAR 表达 CD4 胞外域以重定向 CTL 对抗 HIV 的活性，以及​​刺激 CTL 功能的细胞内 T 细胞信号传导域。这些基因 修饰的 T 细胞靶向病毒生命周期中的关键步骤，与 MHC 呈递无关，靶向 异质病毒，同时避免病毒逃逸的可能性。此外，根据我们的初步研究， 我们将加入强效融合抑制剂 maC46 来保护这些转基因 T 细胞免受感染。在 与之前的研究相比，maC46 将以双顺反子方式整合，而不是共转导。这将 具有更高百分比的同时携带 CD4-CAR 和 maC46 的转导细胞的额外好处。这样的 当适应临床时，一项创新将带来更强的产生转基因 T 细胞的能力 环境。我们假设，当用双顺反子 CD4-CAR 和融合转导时，CMV 特异性 T 细胞 抑制剂，根据其 CMV 特异性，将在体内持续存在，但会免受感染，并且会靶向 残留/重新激活的 HIV+ 细胞。这些实验将是双顺反子载体的首次使用 CD4-CAR 与 maC46 转导到 CMV 特异性 T 细胞中，并显示出对人源化小鼠的拯救， 和恒河猴模型。我们将通过以下具体目标来解决这个假设：1）转换 具有双顺反子 CD4-CAR/maC46 的 CMV 特异性 T 细胞并评估病毒复制的抑制，2) 过继 使用 NSG 小鼠体内转移 CMV 特异性 CD4-CAR/maC46 转导 T 细胞并用 HIV 攻击 感染，3) 将 CMV 特异性 CD4-CAR/maC46 转导 T 细胞过继转移至 SHIV 感染中 恒河猴并与 ART 治疗在血浆病毒载量方面的临床疗效进行比较， 传播性和潜在储存库。由于恒河猴是艾滋病病毒的重要动物模型 发病机制和基因治疗，恒河猴/SHIV模型中转基因T细胞的评估将 快速转化为临床。转基因 T 细胞在缺乏病毒的情况下控制病毒血症的能力 ART，特别是在恒河猴/挑战模型中，将是艾滋病毒治疗和治疗方面的重大进步 将大力推动转基因 T 细胞治疗艾滋病毒/艾滋病的新临床试验。