Modified CMV-specific T cells to Target HIV

修饰 CMV 特异性 T 细胞以靶向 HIV

• 批准号: 8930050
• 负责人: Stephen Edward Braun
• 金额: $ 18.42万
• 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/8930050
• 关键词: AIDS/HIV problem; Acquired Immunodeficiency Syndrome; Adoptive Transfer; Animal Model; Animals; Anti-Retroviral Agents; Antigen Receptors; 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; Mus; 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; Viremia; Virus; Virus Replication; antigen binding; antiretroviral therapy; base; cellular transduction; chimeric antigen receptor; clinically relevant; design; experience; extracellular; fitness; gene therapy; immune function; improved; in vitro activity; in vivo; inhibitor/antagonist; internal control; killings; leukemia; novel; oncology; public health relevance; purge; research study; simian human immunodeficiency virus; tumor; vector; vector-based vaccine

 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：感染人类免疫缺陷病毒-1（HIV-1）导致CD 4 + T细胞耗竭。抗逆转录病毒疗法（ART）虽然在控制血浆病毒血症方面有效， 清除潜在的或持久的水库，以消除感染。最近，研究表明，在50%接种CMV疫苗载体的恒河猴中，病毒复制得到控制，病毒储库减少。他们提出Tem细胞对SIV的持续免疫监视是由持续的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）遗传修饰CMV特异性T细胞，并跟踪对恒河猴中病毒储库的影响。汽车表达CD 4细胞外结构域以重定向针对HIV的CTL活性和细胞内T细胞信号传导结构域以刺激CTL功能。这些“设计者T细胞”（dTc）靶向病毒生命周期中的关键步骤，独立于MHC呈递，靶向异质性病毒，同时避免病毒逃逸的可能性。我们的初步数据（和许多癌症模型）表明，具有多种共刺激信号的汽车（第2代和/或第3代）比原始（第1代）载体具有更强的活性。临床 使用这些高级dTc的试验在减少转移性实体瘤和白血病方面是有效的。此外，基于我们的初步研究，我们还将与有效的融合抑制剂maC 46共转染dTc，以保护这些dTc免受感染。因此，我们假设CMV特异性T细胞，当与CD 4-CAR和融合抑制剂共转导时，基于其CMV特异性将在体内持续存在，但将被保护免受感染，并将靶向残留/再活化的HIV+细胞。在R21阶段，我们将通过在恒河猴模型中测量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，并在SHIV感染后用ART治疗动物。将在淋巴组织储库中测量这些CMV特异性dTc对病毒载量的影响。因此，我们建立了这些关键里程碑：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，以及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CD 4- CAR载体抑制组织中病毒复制的作用。我们将使用内部控制向量， 比较增殖和选择dTc的基础。仅具有TCR的原始⑶ 4-CAR载体是第二代，并且添加来自⑶ 28和4-1BB的胞内信号传导结构域的第三代⑶ 4-CAR将CMV特异性T细胞重定向至HIV，如在肿瘤学中临床上已经实现的。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。以来 恒河猴是研究HIV发病机制和基因治疗的重要动物模型，在恒河猴/SHIV模型中评价dTc将迅速转化为临床。在没有ART的情况下，dTc t控制病毒血症的能力，特别是在恒河猴/攻击模型中，将是HIV治疗的重大进展，并将有力地促进dTc在HIV/AIDS中的新临床试验。