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: 8756661
• 负责人: Jose Alejandro Guevara-Patino
• 金额: $ 16.42万
• 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 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8756661
• 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; 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; public health relevance; 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（由申请人提供）：目前，转移到癌症患者体内后不能持续存在是限制T细胞遗传过继细胞转移有效性的因素。 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DAP 10的胞质信号传导结构域附加到TCR的<$-和<$-链的胞质结构域的末端来修饰人TCR TIL 1383 I（黑素瘤反应性）。我们相信，TCR/DAP 10将在TCR接合时直接激活DAP 10通路，并且这些独特的信号将增强转移的T细胞的存活。初步数据：我们已经在CD 8 + T细胞中证明，通过[天然表达的]活化受体复合物NKG 2D的信号传导增强了记忆发育。我们的数据表明，TCR转导的CD 8 + T细胞中的NKG 2D信号传导增强了其抗肿瘤效力和体内持久性。这些数据表明，通过CD 8 + T细胞中的NKG 2D进行信号传导是克服TCR-Td T细胞有限存活的可行方法。我们将偶联TCR连接和下游NKG 2D信号传导的激活。由于NKG 2D本身不能发出信号，我们将使用DAP 10的信号传导结构域，DAP 10是在CD 8 + T细胞中介导NKG 2D下游信号传导的衔接子分子。我们将通过将DAP 10的胞质信号传导结构域附加到<$-和<$-链的TCR胞质结构域的末端来修饰人TCR TIL 1383 I（酪氨酸酶反应性）。假设：我们假设TCR/DAP 10将在TCR接合时直接激活DAP 10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。 策略：然后将含有TCR/DAP 10构建体的逆转录病毒载体转导入人T细胞。将使用人源化黑素瘤模型检查表达TCR/DAP 10的人T细胞的信号传导途径、存活能力和介导已建立的人癌症消退的能力。免疫缺陷A2/NSG小鼠和人黑素瘤细胞系将用于抗肿瘤实验。 我们将通过以下具体目标SA 1来检验我们的假设。开发TCR/DAP 10构建体以诱导和剖析DAP 10信号传导。SA 2.研究TCR/DAP 10利用的信号通路及其在人T细胞中的细胞后果。SA 3.测试表达TCR/DAP 10的T细胞对抗人黑色素瘤。 我们的策略的意义和创新性在于，这些工程化T细胞将对酪氨酸酶具有特异性，并且它们的TCR的参与将激活DAP 10共刺激途径以及下游TCR信号传导。