Study of TCR Engineering Against Melanoma in Mice

T细胞受体（TCR）工程化的T细胞和造血干细胞（HSC）的过继转移可 潜在地为患者提供大量的肿瘤特异性T细胞，增强基于免疫的治疗， 治疗癌症对这些TCR工程细胞的基本生物学的充分理解将非常有价值地帮助 设计出更有效的疗法在这个项目中，我们建议使用B16黑色素瘤模型进行一项研究。 Pmel TCR工程化的外周CD8 T细胞、CD4 T细胞和HSC的综合研究。为此 为了达到目的，将构建逆转录病毒和慢病毒载体以共同递送编码Pmel TCR的基因 和生物发光报告基因进入靶细胞。TCR工程化T细胞和HSC的体内命运 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 TCR工程化T细胞（特异性Aim1）过继转移产生的免疫力， （特异性目标2）和它们的组合（特异性目标3）。体外TCR转导的各种条件， 将评价受体动物预处理、过继转移和转移后免疫的 有助于有效的治疗。特别地，我们将研究抗肿瘤CD4和CD4 + T细胞之间的协同作用。 CD8 T细胞，以及CD4+CD25+调节性T细胞（TCD4）对过继治疗的影响。我们还将 试图通过共递送一种抗肿瘤药物来增强工程化抗肿瘤T细胞的抗肿瘤效力， 将增强基因与TCR基因一起导入靶细胞。通过这项研究，我们希望 深入了解TCR工程化外周T细胞和HSC的基础生物学， 通过过继转移这些工程细胞产生抗肿瘤免疫力。这种基本的生物学将 通知项目1和项目3中正在进行的临床研究。

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.