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

基本信息

批准号：
7990409
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
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-20 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7990409
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 Generations Granulocyte-Macrophage Colony-Stimulating Factor Growth Histology Human IL2RA gene Image Immunohistochemistry Implant In Situ In Vitro Interleukin 2 Receptor Interleukin-12 Knockout Mice 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

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 Generations Granulocyte-Macrophage Colony-Stimulating Factor Growth Histology Human IL2RA gene Image Immunohistochemistry Implant In Situ In Vitro Interleukin 2 Receptor Interleukin-12 Knockout Mice 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

展开

项目摘要

DESCRIPTION (provided by applicant): 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 imaging 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-3 and the TCR signal arrest has been causally linked to TGF-21 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 non- disrupted pieces of human tumor are surgically implanted into SCID or NOD-SCID/IL2 receptor ? chain null 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.

描述（由申请人提供）：与肿瘤相关的CD4+和CD8+ T细胞具有效应子记忆表型（TEM）内存在于人类非小细胞肺肿瘤和卵巢癌的微环境中，但无法控制肿瘤的进展。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未通过T细胞受体（TCR）响应激活信号。通过阐明在TCR途径中停滞的部位，并定义启动这种逮捕的分子和细胞事件，应该可以设计和测试策略以重新激活TEM原位。预计处理过的肿瘤微环境中TEM的局部重新激活将导致（a）T细胞介导的肿瘤细胞原位杀死肿瘤细胞的杀死，（b）将肿瘤抗原释放到循环中，（c）从现有的tamore sere sere ney或niment hite sere nee nee nee neational sere的肿瘤产生，（c）与现有的肿瘤相邻的肿瘤抗原。使用多光谱成像流式细胞仪，共聚焦显微镜，蛋白质印迹和RTPCR的组合我们的第一个目的是确定在TCR途径中的位置，转导信号被阻断。在相关的第二个目标中，与触发TCR停滞有因果关系的细胞和分子取决于细胞耗竭并添加后退方案，并通过监测所选分子对该调节信号检查点的启动的影响。我们的初步研究将信号检查点的位点定位为PLC-3上游的某个地方，TCR信号停滞与TGF-21有因果关系，从而证明了我们实验方案的可行性和可行性。从这些机械研究中获得的结果将用于目标2和3中，以识别直接或间接在T细胞上起作用的生物活性因素，以逆转其无反应性，并开发和测试脂质体配方，旨在以局部和持续的方式提供这些因素。在最终目标中，评估了每个因素重新激活TEM的治疗功效，并评估诱导局部和系统性的抗肿瘤反应。后者将使用已建立的异种移植模型来完成，其中未破坏的人类肿瘤被手术植入SCID或点头SCID/IL2受体？链无小鼠。在这些异种移植物中，保留了肿瘤微环境，并且相关的白细胞长期对细胞因子信号的响应持续延长。接种后，将负载脂质体负载到异种移植物肿瘤杀伤和TEM反应模式后，以确定每种脂质体制剂的治疗功效。这些研究有望为设计策略的设计奠定基础，这些策略可用于增强我们当前的癌症疫苗临床试验的功效。