Generation of Anti-Melanoma T Cells Derived from Human Embryonic Stem Cells

项目4：从人类胚胎干细胞中培养抗黑色素瘤T细胞 摘要人类免疫系统的基因操作是对抗传染病和癌症的一种有效方法。临床上，目前使用的最常见的基于细胞的治疗方法是造血干细胞移植。将特定的T细胞受体(TCR)稳定地引入干细胞中以产生针对特定抗原的T淋巴细胞的可能性为新的和令人兴奋的终身疫苗和治疗策略打开了大门。人类胚胎干细胞(HESC)和最近被描述的诱导多能干细胞(IPS)具有革新移植技术的潜力。从理论上讲，这些细胞能够分化为体内的任何细胞类型，这为改进细胞替换、器官替换或基因治疗打开了可能性。诱导多能干细胞具有特殊的吸引力，因为从理论上讲，它们可以从单个患者身上获得，因此它们将表达相同的人类白细胞抗原分子，因此它们不会被患者的免疫反应排斥。我们已经证明，hESC可以被诱导分化为T系细胞，并且IPS细胞也具有造血潜力。引入这些细胞的新遗传物质可以在整个造血细胞分化中表达。因此，在免疫系统的遗传操作方面，这些干细胞适应临床应用的潜力尤其真实。使用这些全能干细胞的可能性比使用HSC有许多优势，包括广泛培养的能力和易于基因操作，但我们在控制分化为专门的成熟细胞类型方面存在许多空白。优化和研究hESC和IPS生成功能性T细胞的模型系统的开发将非常有助于提高我们从基因上操纵免疫反应的能力。这项建议将侧重于临床前研究，通过表达针对黑色素瘤的TCRs，从全能干细胞中优化针对人类黑色素瘤的人T淋巴细胞的生成。这些研究将与本计划项目中的其他组成部分相互作用，以提供以下原则：hESC和/或IPS可用于操纵免疫系统，通过追求以下特定目标来特异性靶向癌症：1)从人胚胎和诱导的多能干细胞培养黑色素瘤特异性细胞毒T细胞；2)确定MART-1 TCR的表达如何影响hESC和IPS细胞的胸腺生成潜能；3)开发T细胞特异性表达系统，以优化TCR在hESC和IPS后代中的表达分布。我们希望 与本计划项目的其他组件一起，我们将提供重要的临床前 这些多功能干细胞可以产生癌症特异性T细胞，这种方法可能有治疗黑色素瘤的潜力。

Project 4; Generation of Anti-Melanoma T cells Derived From Human Embryonic Stem Cells Abstract Genetic manipulation of the human immune system is a powerful approach to combating infectious diseases and cancers. Clinically, the most common cell based therapeutic approach currently used is hematopoietic stem cell (HSC) transplantation. The possibility of stably introducing specific T cell receptors (TCRs) into stem cells to generate T lymphocytes targeting specific antigens of choice opens doors to new and exciting life-long vaccine and therapeutic strategies. Human embryonic stem cells (hESC) and the recently-described induced pluripotent stem (IPS) cells, have the potential to revolutionize transplant technology. The theoretical ability of these cells to differentiate into any cell type in the body opens the possibility of improved cell replacement, organ replacement or genetic therapies. Induced pluripotent stem cells have a particular attraction, in that they can theorefically be derived from individual patients and will thus express idenfical HLA molecules, so they would not be rejected by the patient's immune response. We have demonstrated that hESC can be induced to differentiate into cells of the T lineage, and that IPS cells also have hematopoietic potential. New genetic material introduced into these cells can be expressed throughout hematopoietic differenfiafion. Thus the potential for adaptation of these stem cells to clinical use is especially true in regards to genetic manipulation of the immune system. The possibility of using these totipotent stem cells has many advantages over the use of HSC, including ability to culture extensively and ease of genefic manipulafion, however there are many gaps in our knowledge regarding control of differentiation into specialized mature cell types. Development of model systems to optimize and study hESC and IPS generation into funcfional T cells could be very helpful for advancing our ability to genetically manipulate immune responses. This proposal will focus on pre-clinical studies optimizing the generation, from totipotent stem cells, of human T lymphocytes directed to target human melanoma, by virtue of expression of TCRs specific for melanoma anfigens. These studies will interact with other components in this Program Project to provide poof-of-principle that hESC and/or IPS can be used to manipulate the immune system to specifically target cancer by pursuing the following specific aims: 1) Develop melanoma-specific cytotoxic T cells from human embryonic and induced pluripotent stem cells ; 2) Determine how expression of MART-1 TCR influences thymopoietic potential of hESC and IPS cells; 3) Develop T-cell specific expression systems to opfimize distribution of TCR expression in progeny of hESC and IPS. We hope that together with the other components of this Program Project we will provide important pre-clinical informafion that cancer-specific T cells can be generated from these versatile stem cells, and that this approach may have potential for treating melanoma.