TCR/DAP10 chimera as a means to attaining persistent anti-tumor T cell responses

TCR/DAP10 嵌合体作为获得持久抗肿瘤 T 细胞反应的手段

• 批准号: 8918555
• 负责人: Jose Alejandro Guevara-Patino
• 金额: $ 19.71万
• 依托单位: LOYOLA UNIVERSITY CHICAGO
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8918555
• 关键词: Adoptive Cell Transfers; Adoptive Transfer; Antigens; Biological; Bypass; CD4 Positive T Lymphocytes; CD8B1 gene; Cancer Patient; Cell Line; Cell physiology; Chimera organism; Complex; Cytoplasmic Tail; Data; Development; Down-Regulation; Effectiveness; Engineering; Failure; Goals; Grant; Health; Human; Hybrids; Immune; Immune response; Individual; Lead; Ligation; Link; MAPK9 gene; Malignant Neoplasms; Mediating; Melanoma Cell; Memory; Modeling; Monophenol Monooxygenase; Mus; Pathway interactions; Patients; Receptor Activation; Retroviral Vector; Series; Signal Pathway; Signal Transduction; Specificity; T cell response; T-Cell Receptor; T-Lymphocyte; Tail; Testing; cellular engineering; design; in vivo; inhibitor/antagonist; innovation; melanoma; mutant; novel; receptor; research study; response; tumor

DESCRIPTION (provided by applicant): Currently, failure to persist after transfer into cancer patients is a factor that limits the effectiveness of adoptive cell transfer of T cells genetically modified to express a tumor-reactive T cell receptor. The goal of this application is to modify th human TCR TIL 1383I (melanoma-reactive) by appending the cytoplasmic signaling domains of DAP10 to the end of the cytoplasmic domains of the �- and �- chains of the TCR. We believe that the TCR/DAP10 will directly activate the DAP10 pathway upon TCR engagement, and that these unique signals will enhance the survival of transferred T cells. Preliminary data: We have demonstrated in CD8+ T cells that signaling through the [naturally expressed] activation receptor complex NKG2D enhances memory development. Our data show that NKG2D signaling in TCR-transduced CD8+ T cells augments their anti-tumor potency and in vivo persistence. These data argue that signaling through NKG2D in CD8+ T cells is a viable approach to overcoming the limited survival of TCR- Td T cells. We will couple TCR ligation and activation of downstream NKG2D signaling. Because NKG2D is unable to signal by itself, we will use the signaling domain of DAP10, the adaptor molecule that mediates signaling downstream from NKG2D in CD8+ T cells. We will modify the human TCR TIL 1383I (tyrosinase- reactive) by appending the cytoplasmic signaling domains of DAP10 to the end of the TCR cytoplasmic domains of the �- and �- chains. Hypothesis: We hypothesize that the TCR/DAP10 will directly activate the DAP10 stimulatory pathway upon TCR engagement, and that these unique signals will enhance the survival of adoptively transferred T cells. Strategy: The retroviral vector containing TCR/DAP10 construct will then be transduced into human T cells. Human T cells expressing TCR/DAP10 will be examined for their signaling pathways, capacity to survive and ability to mediate the regression of established human cancer using a humanized model of melanoma. Immune deficient A2/NSG mice and human melanoma cell lines will be used for anti-tumor experiments. We will test our hypothesis through the following specific aims SA1. Develop TCR/DAP10 constructs to induce and dissect DAP10 signaling. SA2. Study the signaling pathways utilized by TCR/DAP10 and their cellular consequences in human T cells. SA3. Test T cells expressing TCR/DAP10 against human melanoma. The significance and innovative character of our strategy would be that these engineered T cells will have specificity for tyrosinase, and engagement of their TCR will activate the DAP10 costimulatory pathway as well as downstream TCR signaling.

描述（申请人提供）：目前，转移到癌症患者体内后未能持续存在是限制T细胞过继细胞遗传遗传转移有效性的一个因素 修饰以表达肿瘤反应性 T 细胞受体。本申请的目标是通过将 DAP10 的细胞质信号结构域附加到 β 和 β 细胞质结构域的末端来修饰人类 TCR TIL 1383I（黑色素瘤反应性）。 - TCR 链。我们相信 TCR/DAP10 将在 TCR 参与后直接激活 DAP10 通路，并且这些独特的信号将增强转移 T 的存活。初步数据：我们在 CD8+ T 细胞中证明，通过[自然表达的]激活受体复合物 NKG2D 发出的信号可增强记忆发育。我们的数据表明，TCR 转导的 CD8+ T 细胞中的 NKG2D 信号增强了其抗肿瘤效力。这些数据表明，通过 CD8+ T 细胞中的 NKG2D 信号传导是克服 TCR-Td T 细胞有限存活的可行方法。下游 NKG2D 信号传导的连接和激活 由于 NKG2D 自身无法发出信号，因此我们将使用 DAP10 的信号传导结构域，该接头分子在 CD8+ T 细胞中介导 NKG2D 下游的信号传导。 - 反应性）通过将 DAP10 的细胞质信号结构域附加到 �- 和 的 TCR 细胞质结构域的末端假设：我们认为 TCR/DAP10 在 TCR 参与后会直接激活 DAP10 刺激通路，并且这些独特的信号将增强过继转移的 T 细胞的存活率。然后将其转导到表达 TCR/DAP10 的人类 T 细胞中，并使用人源化方法检查其信号通路、生存能力和介导已建立的人类癌症消退的能力。免疫缺陷 A2/NSG 小鼠和人类黑色素瘤细胞系将用于抗肿瘤实验，通过以下具体目标 SA1 开发 TCR/DAP10 构建体来诱导和剖析 SA2。研究 TCR/DAP10 使用的信号通路及其在人类 T 细胞中的细胞后果。测试表达 TCR/DAP10 的 T 细胞对人类黑色素瘤的意义和作用。我们策略的创新特征是这些工程化 T 细胞将对酪氨酸酶具有特异性，并且其 TCR 的参与将激活 DAP10 共刺激途径以及下游 TCR 信号传导。