Modified CMV-specific T cells to Target HIV

修饰 CMV 特异性 T 细胞以靶向 HIV

• 批准号: 8842407
• 负责人: Stephen Edward Braun
• 金额: $ 19.85万
• 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/8842407
• 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CD4+T细胞耗尽。抗逆转录病毒疗法(ART)虽然能有效控制血浆病毒血症，但不能 清除潜伏的或持久性的储藏物，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，病毒复制得到了控制，病毒库减少了。他们认为，Tem细胞对SIV的持续免疫监视是由持久的CMV载体维持的。为了在体内模拟免疫监测和提高HIV特异性CTL活性，我们将用嵌合抗原受体(CAR)基因修饰CMV特异性T细胞，并观察其对恒河猴病毒库的影响。CARS表达CD4胞外域以重定向抗HIV的CTL活性，并表达胞内T细胞信号域以刺激CTL功能。这些“设计者T细胞”(DTC)针对病毒生命周期中独立于MHC呈递的关键步骤，针对异种病毒，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，带有多个共刺激信号(第二代和/或第三代)的CARS比原始(第一代)载体具有更强的活性。临床 用这些先进的DTC进行的试验在减少转移的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DTC与有效的融合抑制剂maC46共转导，以保护这些DTC免受感染。因此，我们假设，当与CD4-CAR和融合抑制剂共转导CMV特异性T细胞时，基于其CMV特异性，CMV特异性T细胞将在体内持续存在，但将受到保护，不受感染，并将攻击残留/重新激活的HIV+细胞。在R21阶段，我们将通过测量CMV特异性DTC在恒河猴模型中的体内抗HIV活性来确定这些实验的可行性。我们将遵循一个临床方案，并在感染SHV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，我们建立了以下关键里程碑：1)证明了CMV特异性DTC在体内的有效共转导/基因标记，2)测量了CMV特异性DTC在体内的分布和功能CTL活性，3)测量了对血浆和组织病毒血症的影响。在R33阶段，我们将扩大这些研究，以衡量表达CMV特异性DTC的CD4-CAR载体在组织中抑制病毒复制的效果。我们将使用内部控制向量作为 比较DTC增殖和选育的依据。最初的CD_4-CAR载体只有第二代T细胞受体ζ，第三代CD_4-CAR载体增加了CD28和4-1BB的细胞内信号域，将把巨细胞病毒特异性T细胞重定向到HIV，这已经在肿瘤学临床上完成了。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效果。自.以来 恒河猴是研究HIV致病机制和基因治疗的重要动物模型，DTC在恒河猴/SIV模型中的评价将很快进入临床。DTC在缺乏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治疗的一个重大进步，并将有力地推动DTC治疗HIV/AIDS的新的临床试验。