Study of TCR Engineering Against Melanoma in Mice

过继转移T细胞受体(TCR)工程T细胞和造血干细胞(HSCs)可以 潜在地为患者提供大量的肿瘤特异性T细胞，加强基于免疫的治疗 治疗癌症。充分了解这些TCR工程细胞的基本生物学将非常有价值 设计一种更有效的疗法。在这个项目中，我们建议使用B16黑色素瘤模型来进行 PMEL TCR工程外周CDS T细胞、CD4T细胞和HSCs的综合研究为了这个 目的：构建逆转录病毒和慢病毒载体以共同传递编码PMEL TCR的基因。 和一个生物发光记者进入目标细胞。TCR基因工程T细胞和造血干细胞在体内的去向 将在活体动物中使用生物发光成像(BLI)进行实时监测。我们计划研究抗黑色素瘤 过继转移TCR工程T细胞(特异性Aim1)、HSCs产生的免疫 (特异性AIM2)和它们的组合(特异性Aim3)。体外TCR转导的各种条件， 将评估受体动物的预适应、领养转移和移植后免疫 对有效治疗的贡献。特别是，我们将研究抗肿瘤CD 4和CD 4之间的协同作用 CDS T细胞，以及CD4+CD25+调节性T细胞(Tregs)对过继治疗的影响。我们还将 联合应用基因工程抗肿瘤T细胞增强抗肿瘤效应的研究 将增强基因与TCR基因一起导入靶细胞。从拟议的研究来看，我们预计 深入了解TCR工程的外周T细胞和造血干细胞的基本生物学，以及 通过过继转移这些工程细胞产生抗肿瘤免疫。这一基础生物学将 告知项目1和项目3中正在进行的临床调查。

The adoptive transfer of T cell receptor (TCR) engineered T cells and hematopoietic stem cells (HSCs) can potentially provide a patient with a large supply of tumor-specific T cells, enhancing immune-based therapies for cancer. A full understanding of the basic biology of these TCR engineered cells will be very valuable to help design a more potent therapy. In this project, we propose to use the B16 melanoma tumor model to conduct a comprehensive study of the Pmel TCR engineered peripheral CDS T cells, CD4 T cells and HSCs. For this purpose, retroviral and lentiviral vectors will be constructed to co-deliver the genes encoding the Pmel TCR and a bioluminescence reporter into the target cells. The in vivo fate of the TCR-engineered T cells and HSCs will be monitored using Bioluminescence Imaging (BLI) in a live animal in real-time. We plan to study the antimelanoma immunity generated by the adoptive transfer of TCR-engineered T cells (Specific Aim1), HSCs (Specific Aim2), and their combination (Specific Aim3). Various conditions for the in vitro TCR transduction, recipient animal pre-conditioning, adoptive transfer, and post-transfer immunization will be evaluated for their contribution to an effective therapy. In particular, we will study the synergy between the anti-tumor CD4 and CDS T cells, and the influence of the CD4+CD25+ regulatory T cells (Tregs) for adoptive therapy. We will also try to enhance the anti-tumor effectiveness of the engineered anti-tumor T cells by co-delivery of an enhancement gene together with the TCR genes into the target cells. From the proposed study, we expect to gain a thorough understanding of the basic biology of TCR-engineered peripheral T cells and HSCs, and the anti-tumor immunity generated through adoptive transfer of these engineered cells. This basic biology will inform the clinical investigations being conducted in Project 1 and Project 3.