Impact of Highly Active T Lymphocytes from Induced Pluripotent Stem Cells

描述（由申请人提供）：抗原（Ag）特异性CD8+细胞毒性T淋巴细胞（ctl）的过继细胞转移（ACT）是一种很有前途的治疗多种恶性肿瘤的方法。众所周知，分化程度较低的中枢记忆样（称为高度贪婪）ctl是基于act的免疫治疗的最佳群体。然而，由于难以产生与患者靶Ag反应的ctl，使用此类ctl的ACT通常是不可行的。寻找一种产生高度贪婪的ctl的新机制对于成功的基于act的癌症治疗至关重要。长期目标是引发对肿瘤Ags的有效免疫反应，并促进可用于成功预防和治疗人类癌症的疫苗和免疫疗法的开发和应用。本应用的目的是通过开发一种从诱导多能干细胞（iPS）中产生高度渴望的ctl的新方法，并确定iPS细胞衍生的ctl在治疗癌症中的治疗潜力，从而为实现这一目标做出重大贡献。核心假设是，被ag特异性T细胞受体（TCR）基因转导并受到体外Notch信号刺激的iPS细胞能够分化成大量高度渴望的ag特异性ctl。提出这项研究的基本原理是，一旦知道如何从iPS细胞中产生高度贪婪的ag特异性ctl，就有可能通过iPS衍生的ctl的ACT引发有效的抗肿瘤免疫，从而为预防和治疗癌症提供新的创新方法。在初步数据的指导下，这一假设将通过追求两个特定目标来验证：1)从小鼠iPS细胞中产生并表征高度狂热的卵白蛋白（OVA）特异性ctl；2)生成并表征来自人类iPS细胞的高度狂热的黑色素瘤特异性ctl。在第一个目标下，一种已经在体外和体内证明的方法，在申请人手中已经确立为可行的，将用于从小鼠iPS细胞中产生高度活跃的ova特异性ctl。在第二个目标下，与第一个目标中描述的方法类似，将用于从人类iPS细胞中产生高度活跃的黑色素瘤特异性ctl，用于基于act的癌症免疫治疗。本申请中提出的研究是创新的，因为它将开发一种新的方法，从iPS细胞中产生高度贪婪的ag特异性ctl。这种独特的方法可以用于产生高度活跃的肿瘤特异性ctl来治疗癌症，并最终允许开发新的免疫疗法或新疫苗。这项研究具有重要意义，因为(i)除了从根本上推进T细胞生物学领域外，iPS细胞衍生的免疫细胞极有可能用于癌症的预防和治疗干预；（ii）本申请中描述的研究将扩大我们在使用人类和动物干细胞方面的专业知识；（iii）这些研究将为进一步开发来自iPS细胞的高度免疫细胞，并最终产生针对患者和/或癌症的治疗方法提供坚实的基础。

DESCRIPTION (provided by applicant): Adoptive cell transfer (ACT) of antigen (Ag)-specific CD8+ cytotoxic T lymphocytes (CTLs) is a promising treatment for a variety of malignancies. It is known that less-differentiated, central-memory-like (termed highly avid) CTLs are the optimal population for ACT-based immunotherapy. However, ACT with such CTLs is often not feasible due to difficulties in generating these CTLs reactive with the targeted Ag from patients. Finding a new mechanism to generate highly avid CTLs is critical for the successful ACT-based cancer therapy. The long-term goal is to elicit efficient immune responses to tumor Ags and facilitate the development and application of vaccines and immunotherapy that can be used to successfully prevent and treat cancer in humans. The objective in this application is to make a significant contribution to this goal by developing a novel approach to generate highly avid CTLs derived from induced pluripotent stem (iPS) cells, and determining the therapeutic potential of iPS cell-derived CTLs in the treatment of cancer. The central hypothesis is that iPS cells that are transduced with genes of Ag-specific T cell receptor (TCR) and stimulated with in vitro Notch signaling are capable of differentiating into large numbers of highly avid Ag-specific CTLs. The rationale for the proposed research is that, once it is known how to generate highly avid Ag-specific CTLs derived from iPS cells, there is potential to elicit efficient anti-tumor immunity by ACT of iPS-derived CTLs, resulting in new and innovative approaches to the prevention and treatment of cancer. Guided by preliminary data, this hypothesis will be tested by pursuing two specific aims: 1) Generate and characterize highly avid Ovalbumin (OVA)-specific CTLs derived from murine iPS cells; and 2) Generate and characterize highly avid melanoma-specific CTLs derived from human iPS cells. Under the first aim, an already proved in vitro and in vivo approach, which has been established as feasible in the applicant' hand, will be used to generate highly avid OVA-specific CTLs derived from murine iPS cells. Under the second aim, a similar approach as described in the first aim, will be used to generate highly avid melanoma-specific CTLs derived from human iPS cells for ACT-based cancer immunotherapy. The research proposed in this application is innovative, because it will develop a novel approach to generate highly avid Ag-specific CTLs derived from iPS cells. This unique approach can be used for the generation of highly avid tumor-specific CTLs for the treatment of cancer, and will ultimately allow the development of new treatments of immunotherapy or novel vaccines. The proposed research is significant, because (i) the iPS cell-derived immune cells are highly likely to be used in preventive and therapeutic interventions for cancer in addition to fundamentally advancing the fields of T cell biology; (ii) the research described in this application will broaden our expertise regarding the use of human and animal stem cells; (iii) these studies will provide a solid foundation for further development of highly avid immune cells derived from iPS cells, and ultimately for generating patient- and/or cancer-specific therapies. PUBLIC HEALTH RELEVANCE: The proposed research is relevant to public health because it will develop a novel approach to generate highly avid Ag-specific CTLs derived from iPS cells. This unique approach can be used for the generation of highly avid tumor-specific CTLs for the treatment of cancer. Thus, the proposed research is relevant to the part of NIH's mission that conducts and supports research and training with respect to the prevention and treatment of cancer.