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（由申请人证明）：目前，转移到癌症患者中后未能持续存在是在遗传上T细胞转移的AA顾问性 修改以表达DAP10的细胞质信号传导域，并将TCR的链直接激活DAP10途径。在CD8+ T细胞中，通过[自然表达]激活受体NKG2D的信号增强了记忆的发育。克服Will夫妇的TCR连接和下游NKG2D信号的激活，我们将使用DAP10的象征域，该象征域（适配器分子）在CD8+ T细胞中从NKG2D下游信号。 TIL 1383i（酪氨酸酶反应）由AP10的细胞质信号传导结构域到TCR细胞质域的末端。通过传递的T细胞：逆转载体构造TCR/DAP10构建体将使用A2/NSG小鼠的信号途径来检查表达TCR/DAP10黑色素瘤线将用于抗肿瘤实验，我们将通过以下特定目标SA2测试我们的假设。 。