Directed differentiation of HBV-specific CTL from iPSC for cell-based therapies

从 iPSC 中定向分化 HBV 特异性 CTL，用于基于细胞的治疗

• 批准号: 8772776
• 负责人: Jianxun Jim Song
• 金额: $ 24.05万
• 依托单位: PENNSYLVANIA STATE UNIV HERSHEY MED CTR
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-18 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8772776
• 关键词: Adoptive Cell Transfers; Antigens; Antiviral Therapy; Apoptosis; CD8B1 gene; Cell Therapy; Cells; Chronic Hepatitis B; Communicable Diseases; Cytotoxic T-Lymphocytes; Data; Development; Foundations; Genomic DNA; Goals; HLA-A2.1; Hematopoietic; Hematopoietic stem cells; Hepatitis B Virus; Immunotherapeutic agent; Immunotherapy; In Vitro; Knowledge; Laboratories; Ligands; Malignant Neoplasms; Malignant neoplasm of liver; Memory; Modeling; Mus; Patients; Peripheral Blood Mononuclear Cell; Phenotype; Pluripotent Stem Cells; Population; Primary carcinoma of the liver cells; Property; Publishing; RNA-Directed DNA Polymerase; Receptor Cell; Research; Satellite Viruses; Signal Transduction; Somatic Cell; Source; Specificity; System; T memory cell; T-Cell Receptor; T-Cell Receptor Genes; T-Lymphocyte; Testing; Therapeutic Uses; Transfection; Transgenic Mice; Viral Antigens; Virus; Virus Diseases; Virus Replication; anti-hepatitis B; base; c-myc Genes; cytokine; embryonic stem cell; in vivo; induced pluripotent stem cell; infectious disease treatment; innovation; insight; notch protein; prevent; programs; public health relevance; tumor; tumor growth

DESCRIPTION (provided by applicant): Virus-specific T cells capable of controlling hepatitis B virus (HBV) and eliminating hepatocellular carcinoma (HCC) expressing HBV antigen (Ag) are deleted or dysfunctional in patients with chronic HBV infection. Current antiviral therapy targets the virus reverse transcriptase and rarely establishes immunological control over HBV replication. Adoptive cell transfer (ACT) of HBV-specific CD8+ cytotoxic T lymphocytes (CTL) is an highly promising treatment for chronic HBV infection and HBV-associated HCC. Naive or central memory T cell-derived effector CTL - the "right" or "highly reactive" CTL are the optimal populations for ACT-based immunotherapy, because these cells have a high proliferative potential, are less prone to apoptosis than terminally differentiated cells and have a higher ability to respond to homeostatic cytokines. However, such ACT is often not feasible due to difficulties in obtaining sufficient numbers of highly reactive CTL from patients. The long-term goal of our research program is to develop and optimize strategies for utilizing pluripotent stem cells (PSC) expressing Ag-specific T cell receptor (TCR) as a source of highly reactive CTL for cell-based therapies. The objective in this application is to determine the fundamental properties of HBV-specific CTL from induced PSC (i.e., iPSC-CTL) genetically modified with HBV-specific TCR, for potential immunotherapeutic development against chronic HBV infection and HBV-associated HCC. The central hypothesis is that HBV-specific iPSC-CTL developed by our in vitro differentiation system are naive monoclonal HBV-specific CD8+ T cells, which can be used to generate highly reactive HBV-specific CTL for cell-based therapies. Guided by published results and preliminary data from the applicant's laboratory, this hypothesis will be tested by pursuing two specific aims: 1) to define the fundamental properties of HBV-specific iPSC-CTL that relate to their anti-HBV activity; and 2) to determine the anti-HBV activity of HBV-specific iPSC- CTL in HBV-bearing mice. Under the first aim, in vitro and in vivo approaches, which have been established as feasible in the applicant's laboratory, will be used. The fundamental properties of HBV-specific iPSC-CTL regarding phenotype, specificity and function will be defined. Under the second aim, using murine models of HBV infection and HBV-associated tumor, the efficacy of HBV-specific iPSC-CTL for cell-based therapies will be determined. The approach is innovative, because HBV-specific iPSC-CTL genetically modified with HBV- specific TCR for cell-based therapies of chronic HBV infection and HBV-associated HCC has not been previously explored. The proposed research is significant, because it will provide new insight and strategies for generating highly reactive virus-specific PSC-T cells and in so doing drive forward use of therapeutic T cells for the treatments of infectious diseases and cancers.

描述（由申请人提供）：能够控制丙型肝炎病毒（HBV）的病毒特异性T细胞并消除表达HBV抗原（AG）的肝细胞癌（HCC）的肝细胞癌（HCC）在患有慢性HBV感染的患者中被删除或功能障碍。当前的抗病毒疗法靶向病毒逆转录酶，很少建立对HBV复制的免疫控制。 HBV特异性CD8+细胞毒性T淋巴细胞（CTL）的收养细胞转移（ACT）是慢性HBV感染和与HBV相关的HCC的高度有前途的治疗方法。幼稚或中央记忆T细胞衍生的效应子CTL-“右”或“高反应性” CTL是基于ACT的免疫疗法的最佳人群，因为这些细胞具有很高的增殖潜力，比终末分化细胞易于凋亡，并且对稳态细胞因子的反应能力更高。但是，由于难以从患者那里获得足够数量的高反应性CTL，因此这种行为通常是不可行的。我们的研究计划的长期目标是开发和优化使用表达Ag特异性T细胞受体（TCR）的多能干细胞（PSC）作为基于细胞疗法的高反应性CTL的来源。本应用程序中的目的是确定来自诱导的PSC（即IPSC-CTL）遗传使用HBV特异性TCR的HBV特异性CTL的基本特性，以针对慢性HBV感染和HBV相关HCC进行潜在的免疫治疗性发育。中心假设是，由我们的体外分化系统开发的HBV特异性IPSC-CTL是天真的单克隆HBV特异性CD8+ T细胞，可用于基于细胞的疗法生成高反应性HBV特异性CTL。在已发布的结果和申请人实验室的初步数据的指导下，该假设将通过追求两个具体目标来检验：1）定义与其抗HBV活性有关的HBV特异性IPSC-CTL的基本特性； 2）确定HBV特异性IPSC-CTL在含HBV的小鼠中的抗HBV活性。在第一个目的下，将使用在申请人实验室可行的体外和体内方法。 HBV特异性IPSC-CTL关于表型，特异性和功能的基本特性将被定义。在第二个目标下，使用HBV感染和HBV相关肿瘤的鼠模型，将确定HBV特异性IPSC-CTL对基于细胞的疗法的疗效。该方法具有创新性，因为HBV特异性的IPSC-CTL用HBV-特异性TCR进行了基于慢性HBV感染和HBV相关HCC的基于细胞的TCR的基因修饰。拟议的研究非常重要，因为它将提供新的见识和策略，以产生高反应性病毒特异性的PSC-T细胞，因此可以推动前往治疗性T细胞用于治疗传染病和癌症的治疗。