Modified CMV-specific T cells to Target HIV

修饰 CMV 特异性 T 细胞以靶向 HIV

• 批准号: 9177838
• 负责人: Stephen Edward Braun
• 金额: $ 8.66万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-19 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9177838
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Adoptive Transfer; Animal Model; Animals; Anti-Retroviral Agents; Antigens; Back; Binding; Biodistribution; CD28 gene; CD4 Positive T Lymphocytes; CTL assay; Cancer Model; Cells; Cessation of life; Chimeric Proteins; Clinic; Clinical; Clinical Trials; Cytomegalovirus; Cytomegalovirus Vaccines; Data; Disease; Down-Regulation; Effector Cell; Extracellular Domain; Generations; Genes; Goals; HIV; HIV Seropositivity; HIV-1; Homing; Human; Immunologic Monitoring; In Vitro; Infection; Lead; Life; Life Cycle Stages; Link; Lymphoid Tissue; MHC Class I Genes; Macaca mulatta; Measures; Mediating; Membrane; Methods; Modeling; Mutation; Neoplasm Metastasis; Pathogenesis; Pathologic Processes; Patients; Peripheral; Pharmacotherapy; Phase; Phenotype; Plasma; Protocols documentation; Regimen; Residual state; Retroviral Vector; SIV; Signal Transduction; Solid Neoplasm; Specificity; Study models; T cell response; T-Cell Depletion; T-Cell Receptor; T-Lymphocyte; Therapeutic; Therapy Evaluation; Tissues; Transduction Gene; Translating; Tumor Antigens; Vaccinated; Viral; Viral Load result; Viral Physiology; Viral reservoir; Viremia; Virus; Virus Replication; antigen binding; antiretroviral therapy; base; cellular transduction; chimeric antigen receptor; clinically relevant; design; experience; extracellular; gene therapy; immune function; improved; in vitro activity; in vivo; inhibitor/antagonist; internal control; killings; leukemia; mouse model; novel; oncology; public health relevance; purge; research study; simian human immunodeficiency virus; tumor; vector; vector-based vaccine; viral fitness

 DESCRIPTION: Infection with human immunodeficiency virus-1 (HIV-1) results in CD4+ T cell depletion. Antiretroviral therapy (ART), although effective in controlling plasma viremia, does not purge the latent or persistent reservoirs necessary to eliminate the infection. 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 immuno surveillance of SIV by Tem cells is maintained by the persistent CMV vectors. To mimic the immuno surveillance 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 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 "designer T cells" (dTc) target a critical step in the viral life cycle independent of MHC presentation, targeting heterogeneous viruses while avoiding the potential for viral escape. Our preliminary data (and many cancer models) show that CARs with multiple co-stimulatory signals (2nd and/or 3rd generation) have stronger activity than original (1st generation) vectors. Clinical trials with these advanced dTc have been efficacious in reducing metastatic solid tumors and leukemia. Additionally, based our preliminary studies, we will also co-transduce dTc with the potent fusion inhibitor maC46 to protect these dTc from infection. Therefore, we hypothesize that CMV-specific T cells, when co-transduced with CD4-CAR and the fusion inhibitor, will persist in vivo based on their CMV specificity, but will be protected from infection and will targe residual/reactivated HIV+ cells. In the R21 phase, we will determine the feasibility of these experiments by measuring the in vivo anti-HIV activity of CMV-specific dTc in rhesus macaque model. We will follow a clinical scenario and treat animals with ART after SHIV infection. The effects of these CMV-specific dTc on viral load will be measured in lymphoid tissue reservoirs. Therefore, we established these critical milestones: 1) demonstrate efficient co- transduction/gene marking of CMV-specific dTc in vivo, 2) measure the distribution and functional CTL activity in vivo, and 3) measure the effects on plasma and tissue viremia. In the R33 phase, we will expand these studies to measure the effect of CMV-specific dTc expressing CD4- CAR vectors to inhibit viral replication in tissues. We will use an internal control vector as the basis for comparing proliferation and selection of the dTc. The original CD4-CAR vector with only the TCRζ the 2nd generation, and the 3rd generation CD4-CAR adding intracellular signaling domains from CD28 and 4-1BB will redirect the CMV-specific T cells towards HIV, as have been accomplished clinically in oncology. We will compare the persistence and homing, CTL activity, the selective expansion, and ultimately, the efficacy of the different vectors. Since rhesus macaque is an important animal model for HIV pathogenesis and gene therapy, the evaluation of dTc in rhesus/SHIV model will translate quickly into the clinic. The ability of dTc t 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 dTc in HIV/AIDS.

 描述：人类免疫缺陷病毒 1 (HIV-1) 感染会导致 CD4+ T 细胞耗竭。抗逆转录病毒治疗（ART）虽然可以有效控制血浆病毒血症，但不能 清除消除感染所必需的潜在或持久的储存库。最近，研究表明，50% 接种 CMV 疫苗载体的恒河猴可控制病毒复制并减少病毒储存。他们提出，Tem 细胞对 SIV 的连续免疫监视是由持久性 CMV 载体维持的。为了模拟免疫监视并增加体内 HIV 特异性 CTL 活性，我们将用嵌合抗原受体 (CAR) 对 CMV 特异性 T 细胞进行基因修饰，并跟踪对恒河猴病毒库的影响。 CAR 表达 CD4 胞外结构域以重定向针对 HIV 的 CTL 活性，并表达胞内 T 细胞信号传导结构域以刺激 CTL 功能。这些“设计 T 细胞”(dTc) 靶向病毒生命周期中独立于 MHC 呈递的关键步骤，靶向异质病毒，同时避免病毒逃逸的可能性。我们的初步数据（以及许多癌症模型）表明，具有多个共刺激信号（第二代和/或第三代）的 CAR 比原始（第一代）载体具有更强的活性。临床 使用这些先进的 dTc 进行的试验可有效减少转移性实体瘤和白血病。此外，根据我们的初步研究，我们还将与强效融合抑制剂maC46共转导dTc，以保护这些dTc免受感染。因此，我们假设 CMV 特异性 T 细胞在与 CD4-CAR 和融合抑制剂共转导时，将根据其 CMV 特异性在体内持续存在，但会受到保护免受感染，并会针对残留/重新激活的 HIV+ 细胞。在R21阶段，我们将通过测量恒河猴模型中CMV特异性dTc的体内抗HIV活性来确定这些实验的可行性。我们将遵循临床情况，在 SHIV 感染后使用 ART 治疗动物。这些 CMV 特异性 dTc 对病毒载量的影响将在淋巴组织储存库中进行测量。因此，我们建立了这些关键的里程碑：1）证明体内 CMV 特异性 dTc 的有效共转导/基因标记，2）测量体内分布和功能性 CTL 活性，3）测量对血浆和组织病毒血症的影响。在R33阶段，我们将扩展这些研究，以测量CMV特异性dTc表达CD4-CAR载体抑制组织中病毒复制的效果。我们将使用内部控制向量作为 比较 dTc 的增殖和选择的基础。最初的第二代 CD4-CAR 载体仅包含 TCRz，而第三代 CD4-CAR 添加了来自 CD28 和 4-1BB 的细胞内信号结构域，将把 CMV 特异性 T 细胞重定向到 HIV，正如肿瘤学临床中所实现的那样。我们将比较不同载体的持久性和归巢、CTL 活性、选择性扩增以及最终的功效。自从 恒河猴是HIV发病机制和基因治疗的重要动物模型，恒河猴/SHIV模型中dTc的评估将很快转化为临床。 dTc 在没有 ART 的情况下控制病毒血症的能力，特别是在恒河猴/挑战模型中，将是 HIV 治疗的重大进步，并将大力推动 dTc 在 HIV/AIDS 中的新临床试验。