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.

项目摘要 人类免疫缺陷病毒1（HIV-1）迄今已杀死了3500万人，并感染了200万人 每年的人。 HIV感染导致CD4+ T细胞的耗竭，导致免疫功能低下 状态和高血浆病毒负荷与高透气性相关。抗逆转录病毒疗法（ART），尽管 有效控制血浆病毒血症和传播，不会清除潜在或持续的储层 消除感染所必需的。鉴于违规，社会经济障碍和跟进的损失是 成功的艺术疗法的常见障碍，必须发现提供一生的治疗剂 抑制病毒载荷和病毒储层的耗竭。最近，研究表明了病毒的控制 用CMV疫苗载体接种疫苗的50％的恒河猴的复制和降低病毒储量。他们 提出，TEM细胞对SIV的连续免疫监视是由持续的CMV维持的 向量。为了模仿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，并遵循对病毒储层的影响 人源化的小鼠和恒河猕猴。汽车将CD4细胞外域表示为重定向CTL 针对HIV的活性和细胞内T细胞信号传导域，以刺激CTL功能。这些在遗传上 修饰的T细胞针对病毒生命周期中的关键步骤，独立于MHC表现，针对性 异质病毒，同时避免了病毒逃生的潜力。此外，基于我们的初步研究 我们将包括有效的融合抑制剂MAC46，以保护这些转基因T细胞免受感染。在 与先前的研究形成鲜明对比的是，MAC46将以双科性合并，而不是共转导的。这会 具有较高百分比携带CD4型和MAC46的转导细胞的额外好处。这样的 当适应临床时，创新将导致更大的能力产生转基因的T细胞 环境。我们假设用双丝cd4型和融合转导的CMV特异性T细胞 抑制剂将根据其CMV的特异性持续存在，但会受到保护，并将靶向 残留/重新激活的HIV+细胞。这些实验将是掺入的双分散矢量的首次使用 带有MAC46的CD4卡车转导至CMV特异性T细胞，并显示了两种人源性小鼠的拯救 和恒河猕猴模型。我们将通过以下特定目的解决这一假设：1）转导 CMV特异性T细胞，具有双分散性CD4-CAR/MAC46并评估病毒复制的抑制作用，2） 使用NSG小鼠转移CMV特异性CD4-CAR/MAC46在体内转导的T细胞，并与HIV挑战 感染和3）通过过继转移CMV特异性CD4-CAR/MAC46将T细胞转移到感染的SHIV中 恒河猴和与血浆病毒载量有关的艺术疗法的临床疗效进行比较， 传播性和潜在储层。由于猕猴是两种艾滋病毒的重要动物模型 发病机理和基因疗法，恒河类/SHIV模型中转基因T细胞的评估将 迅速转化为诊所。在没有基因修饰的T细胞控制病毒血症的能力的情况下 艺术，尤其是在恒河主/挑战模型中，将是艾滋病毒治疗和 将强烈促进针对HIV/AIDS中转基因T细胞的新临床试验。