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：
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
项目类别：
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

项目摘要

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 CD4+ 和 CD8+ T 细胞存在于人类非小细胞肺癌和卵巢癌的微环境，但未能控制肿瘤进展。我们的数据表明，这些细胞对肿瘤无反应的部分原因是其未能通过 T 细胞受体 (TCR) 响应激活信号。通过阐明逮捕地点在 TCR 途径中，并通过定义引发这种停滞的分子和细胞事件，它应该是可以设计和测试原位重新激活 Tem 的策略。局部重新激活 Tem 治疗后的肿瘤微环境预计会导致 (a) T 细胞介导的原位肿瘤细胞杀伤，(b) 肿瘤抗原释放到循环中，(c) 产生全身抗肿瘤免疫力，以及 (d) T 细胞识别并根除邻近或远离初始治疗部位的现有肿瘤肿瘤部位。结合使用多光谱成像流式细胞术、共聚焦显微镜、蛋白质印迹对于 rtPCR，我们的首要目标是确定 TCR 通路中转导信号被阻断的位置。在一个相关的第二个目标是确定与触发 TCR 停滞有因果关系的细胞和分子通过细胞消耗和添加回方案以及通过监测选定分子对启动的影响这个监管信号检查点。我们的初步研究已经确定了信号检查点的位置发生在 PLC-γ 上游的某个地方，并且 TCR 信号停滞与 TGF-β1 存在因果关系证明我们的实验方案的可行性和可行性。从这些得到的结果目标 2 和 3 将利用机制研究来确定直接或间接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旨在以局部和持续的方式在体内传递这些因子。在最终目标中每个因子原位重新激活 Tem 并诱导局部和全身抗肿瘤的治疗效果响应进行评估。后者将使用已建立的异种移植模型来完成，其中不间断通过手术将人类肿瘤碎片植入 SCID 或 NOD-SCID/IL2 受体 γ 链缺失小鼠体内。在这些异种移植物中，肿瘤微环境得以保留，并且肿瘤相关白细胞仍然存在长时间存活并可预测地对细胞因子信号作出反应。接种后监测负载因子的脂质体进入异种移植物的肿瘤杀灭和TEM反应模式以确定每种脂质体制剂的治疗效果。这些研究预计将为设计可用于提高我们当前癌症疫苗接种临床试验效果的策略。