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（申请人提供）：能够控制乙型肝炎病毒（HBV）并消除表达 HBV 抗原（Ag）的肝细胞癌（HCC）的病毒特异性 T 细胞在慢性 HBV 感染患者中缺失或功能障碍。目前的抗病毒治疗以病毒逆转录酶为目标，很少建立对乙型肝炎病毒复制的免疫控制。 HBV 特异性 CD8+ 细胞毒性 T 淋巴细胞 (CTL) 的过继细胞移植 (ACT) 是治疗慢性 HBV 感染和 HBV 相关 HCC 的一种非常有前景的治疗方法。初始或中枢记忆 T 细胞衍生的效应 CTL——“正确”或“高反应性”CTL 是基于 ACT 的免疫疗法的最佳群体，因为这些细胞具有高增殖潜力，比终末分化细胞更不易凋亡，并且对稳态细胞因子有更高的反应能力。然而，由于难以从患者那里获得足够数量的高反应性 CTL，这种 ACT 通常是不可行的。我们研究计划的长期目标是开发和优化利用表达 Ag 特异性 T 细胞受体 (TCR) 的多能干细胞 (PSC) 作为细胞疗法的高反应性 CTL 来源的策略。本申请的目的是确定用 HBV 特异性 TCR 进行基因修饰的诱导 PSC（即 iPSC-CTL）中的 HBV 特异性 CTL 的基本特性，以用于针对慢性 HBV 感染和 HBV 相关 HCC 的潜在免疫治疗开发。核心假设是，我们的体外分化系统开发的 HBV 特异性 iPSC-CTL 是初始单克隆 HBV 特异性 CD8+ T 细胞，可用于生成高反应性 HBV 特异性 CTL，用于基于细胞的治疗。在已发表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进行检验：1）定义与抗乙肝病毒活性相关的乙肝病毒特异性 iPSC-CTL 的基本特性； 2) 确定 HBV 特异性 iPSC-CTL 在携带 HBV 的小鼠中的抗 HBV 活性。根据第一个目标，将使用申请人实验室已确定可行的体外和体内方法。将定义 HBV 特异性 iPSC-CTL 在表型、特异性和功能方面的基本特性。第二个目标是使用 HBV 感染和 HBV 相关肿瘤的小鼠模型，确定 HBV 特异性 iPSC-CTL 用于细胞疗法的功效。该方法具有创新性，因为之前尚未探索过用 HBV 特异性 TCR 基因修饰的 HBV 特异性 iPSC-CTL 用于慢性 HBV 感染和 HBV 相关 HCC 的细胞疗法。拟议的研究意义重大，因为它将为生成高反应性病毒特异性 PSC-T 细胞提供新的见解和策略，从而推动治疗性 T 细胞在传染病和癌症治疗中的应用。