Re-activating Memory T Cells in the Microenvironment of Human Tumors

重新激活人类肿瘤微环境中的记忆 T 细胞

• 批准号: 8196768
• 负责人: RICHARD B BANKERT
• 金额: $ 33.54万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-20 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8196768
• 关键词: Address; Antibodies; Antigens; Back; Binding; Blood Circulation; CD28 gene; CD3 Antigens; CD8B1 gene; Cancer Patient; Cell Nucleus; Cells; Clinical Trials; Confocal Microscopy; Cytokine Signaling; Cytosol; Data; Distant; Drug Formulations; Event; Failure; Fibroblasts; Flow Cytometry; Foundations; Gene Expression Profile; Generations; Granulocyte-Macrophage Colony-Stimulating Factor; Growth; Histology; Human; IL2RA gene; Immunohistochemistry; Implant; In Situ; In Vitro; Interleukin 2 Receptor; Interleukin-12; Lead; Leukocytes; Link; Liposomes; Location; Lung Neoplasms; Lymphocyte; Malignant Neoplasms; Mediating; Membrane; Memory; Molecular; Monitor; Mus; Ovarian Carcinoma; Pathway interactions; Patients; Pattern; Phenotype; Phosphotransferases; Preparation; Protocols documentation; Receptor Signaling; Regulatory T-Lymphocyte; Serum; Signal Transduction; Signal Transduction Pathway; Site; Small Interfering RNA; T Cell Receptor Signaling Pathway; T memory cell; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Therapeutic; Tissues; Treatment Efficacy; Tumor Antigens; Tumor Immunity; Vaccination; Western Blotting; Xenograft Model; Xenograft procedure; base; blocking factor; cancer cell; cell type; chemokine; crosslink; cytokine; design; in vivo; inhibitor/antagonist; killings; neoplastic cell; ovarian neoplasm; peripheral blood; phosphatase inhibitor; prevent; response; senescence; transcription factor; tumor; tumor progression; tumor xenograft

Tumor-associated CD4+ and CD8+ T cells with an effector memory phenotype (Tem) are present within the microenvironment of human non-small cell lung tumors and ovarian carcinomas, but fail to control tumor progression. Our data have established that the non-responsiveness of these cells to the tumor is due in part to their failure to respond to activation signals via the T cell receptor (TCR). By elucidating the site of the arrest in the TCR pathway and by defining the molecular and cellular events that initiate this arrest it should be possible to design and test strategies to re-activate the Tem in situ. The local re-activation of the Tem within the treated tumor microenvironment is expected to result in, (a) T cell mediated killing of tumor cells in situ, (b) release of tumor antigens into the circulation, (c) generation of a systemic anti-tumor immunity, and (d) T cell recognition and eradication of existing tumors at sites that are adjacent to or distant from the initially treated tumor site. Using a combination of multispectral immaging flow cytometry, confocal microscopy, western blot and rtPCR our first aim is to determine where in the TCR pathway the transduction signal is blocked. In a related second aim the cells and molecules that are causally linked to triggering the TCR arrest are determined by cell depletion and add back protocols and by monitoring the effects of selected molecules on the initiation of this regulatory signaling checkpoint. Our preliminary studies have localized the site of the signaling checkpoint to occur somewhere upstream of PLC-γ and the TCR signal arrest has been causally linked to TGF-β1 thereby demonstrating the feasibility and viability of our experimental protocols. The results obtained from these mechanistic studies will be utilized in aims 2 and 3 to identify biologically active factors that act directly or indirectly on the T cells to reverse their non-responsiveness, and to develop and test liposome formulations that are designed to deliver these factors in a local and sustained fashion in vivo. In the final aim the therapeutic efficacy of each factor for re-activating Tem in situ and for inducing a local and systemic anti-tumor response is evaluated. The latter is to be accomplished using an established xenograft model in which nondisrupted pieces of human tumor are surgically implanted into SCID or NOD-SCID/IL2 receptor γ chainnull mice. In these xenografts the tumor microenvironment is preserved and the tumor- associated leukocytes remain viable and predictably responsive to cytokine signals for prolonged periods. Following the inoculation of the factor loaded liposomes into the xenografts tumor killing and Tem response patterns are monitored to determine the therapeutic efficacy of each liposomal preparation. These studies are expected to lay the foundation for the design of strategies that can be used to enhance the efficacy of our current cancer vaccination clinical trials.

具有效应记忆表型（Tem）的肿瘤相关CD 4+和CD 8 + T细胞存在于肿瘤细胞内。 人非小细胞肺癌和卵巢癌的微环境，但未能控制肿瘤 进展我们的数据已经确定，这些细胞对肿瘤的无反应性部分是由于 它们无法通过T细胞受体（TCR）对激活信号作出反应。通过说明逮捕地点 在TCR通路中，通过定义启动这种停滞的分子和细胞事件， 我们可以设计和测试策略，重新激活TEM原位。局部重新激活TEM， 预期经处理的肿瘤微环境导致（a）原位T细胞介导的肿瘤细胞杀伤，（B） 肿瘤抗原释放到循环中，（c）产生全身性抗肿瘤免疫，和（d）T细胞 识别和根除邻近或远离初始治疗部位的现有肿瘤 肿瘤部位。使用多光谱成像流式细胞术，共聚焦显微镜，蛋白质印迹 和rtPCR，我们的第一个目标是确定在TCR途径中转导信号被阻断的位置。中 相关的第二个目的是确定与触发TCR停滞有因果联系的细胞和分子 通过细胞耗竭和添加回方案以及通过监测所选分子对细胞增殖的启动的影响， 这个调节信号检查点我们的初步研究已经定位了信号检查点的位置 发生在PLC上游的某个地方-并且TCR信号停滞与TGF- 1有因果关系， 证明了我们的实验方案的可行性和可行性。从这些获得的结果 机制研究将用于目标2和3，以确定直接或间接起作用的生物活性因子。 间接作用于T细胞，以逆转其无反应性，并开发和测试脂质体制剂 其被设计为在体内以局部和持续的方式递送这些因子。在最后的目标中， 每种因子原位再活化Tem和诱导局部和全身抗肿瘤的治疗功效 响应进行了评估。后者将使用已建立的异种移植模型来完成，在该模型中，未破坏的 将人肿瘤块手术植入SCID或NOD-SCID/IL 2受体链缺失小鼠。 在这些异种移植物中，肿瘤微环境得以保留，肿瘤相关白细胞得以保留。 存活的并且可预测地对细胞因子信号响应延长的时间段。接种后， 监测异种移植物肿瘤杀伤和Tem响应模式以确定 每种脂质体制剂的治疗功效。这些研究有望为 设计可用于提高我们目前癌症疫苗临床试验效力的策略。