Impact of Highly Active T Lymphocytes from Induced Pluripotent Stem Cells

描述（由申请方提供）：抗原（Ag）特异性CD 8+细胞毒性T淋巴细胞（CTL）的连续细胞转移（ACT）是一种有前景的治疗多种恶性肿瘤的方法。众所周知，分化程度较低的中央记忆样（称为高亲和力）CTL是基于ACT的免疫疗法的最佳群体。然而，由于难以产生与来自患者的靶向Ag反应的这些CTL，使用此类CTL的ACT通常是不可行的。寻找一种新的机制来产生高亲和力的CTL是成功的基于ACT的癌症治疗的关键。长期目标是引发对肿瘤抗原的有效免疫应答，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。本申请的目的是通过开发一种新的方法来产生源自诱导多能干（iPS）细胞的高度亲合力的CTL，并确定源自iPS细胞的CTL在癌症治疗中的治疗潜力，从而对该目标做出重大贡献。核心假设是，用Ag特异性T细胞受体（TCR）基因转导并用体外Notch信号转导刺激的iPS细胞能够分化成大量高度亲合力的Ag特异性CTL。这项研究的基本原理是，一旦知道如何从iPS细胞产生高度亲合力的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，从而产生预防和治疗癌症的新的创新方法。在初步数据的指导下，将通过追求两个特定目标来测试该假设：1）产生和表征源自鼠iPS细胞的高度亲合力卵清蛋白（OVA）特异性CTL;和2）产生和表征源自人iPS细胞的高度亲合力黑素瘤特异性CTL。在第一个目标下，已经证明的体外和体内方法（其在申请人手中已经被确立为可行的）将用于产生源自鼠iPS细胞的高度亲合力的OVA特异性CTL。在第二个目标下，将使用与第一个目标中所述类似的方法来产生源自人iPS细胞的高度亲合力的黑素瘤特异性CTL，用于基于ACT的癌症免疫疗法。本申请中提出的研究具有创新性，因为它将开发一种新的方法来产生源自iPS细胞的高度亲和力的银特异性CTL。这种独特的方法可用于产生高度亲和力的肿瘤特异性CTL，用于治疗癌症，并最终允许开发新的免疫疗法或新型疫苗。所提出的研究是重要的，因为（i）除了从根本上推进T细胞生物学领域之外，iPS细胞衍生的免疫细胞极有可能用于癌症的预防性和治疗性干预;（ii）本申请中描述的研究将拓宽我们关于人类和动物干细胞使用的专业知识;（iii）这些研究将为进一步开发来源于iPS细胞的高亲和力免疫细胞，并最终产生患者和/或癌症特异性疗法提供坚实的基础。

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.