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: 7353638
• 负责人: RICHARD B BANKERT
• 金额: $ 37.01万
• 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 2012-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7353638
• 关键词: Address; Antibodies; Antigens; Back; Binding; Blood Circulation; CD28 gene; CD3 Antigens; CD8B1 gene; Cancer Patient; Cell Nucleus; Cells; Clinical Trials; Condition; 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; Image; Immunohistochemistry; Implant; In Situ; In Vitro; Interleukin 2 Receptor; Interleukin-12; Knockout Mice; Lead; Leukocytes; Link; Liposomes; Localized; Location; Lung Neoplasms; Lymphocyte; Malignant Neoplasms; Mediating; Membrane; Memory; Molecular; Monitor; Mus; Numbers; Ovarian Carcinoma; Pathway interactions; Patients; Pattern; Phenotype; Phosphotransferases; Preparation; Protocols documentation; Receptor Signaling; Research; 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

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. Memory T cells that are present within and surrounding cancer patients' tumors have the potential to recognize and kill tumor cells, but they often fail to control the growth and spread of the cancer cells in these patients. This failure is due in part to a non-responsive or quiescent state of the memory cells. We are investigating the reason for this quiescent state, designing strategies to reverse this non-responsiveness, and testing the therapeutic benefits of re-activating the tumor-associated memory T cells.

描述（由申请人提供）：具有效应记忆表型（Tem）的肿瘤相关CD4+和CD8 + T细胞存在于人非小细胞肺肿瘤和卵巢癌的微环境中，但不能控制肿瘤进展。我们的数据已经确定，这些细胞对肿瘤的非反应性部分是由于它们未能通过T细胞受体（TCR）对激活信号作出反应。通过阐明TCR途径中的停滞位点并通过定义启动该停滞的分子和细胞事件，应该可以设计和测试原位重新激活Tem的策略。预期在治疗的肿瘤微环境中Tem的局部再活化导致（a）T细胞介导的原位杀伤肿瘤细胞，（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细胞识别和根除在邻近或远离最初治疗的肿瘤部位的部位处的现有肿瘤。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的组合，我们的第一个目标是确定在TCR通路中的转导信号被阻断。在相关的第二个目的中，与触发TCR停滞有因果联系的细胞和分子通过细胞耗竭和加回方案以及通过监测所选分子对该调节信号传导检查点的起始的影响来确定。我们的初步研究已经将信号检查点的位点定位于PLC-3上游的某处，并且TCR信号停滞与TGF-21有因果关系，从而证明了我们的实验方案的可行性和可行性。从这些机制研究中获得的结果将用于目标2和3，以鉴定直接或间接作用于T细胞以逆转其非反应性的生物活性因子，并开发和测试旨在以局部和持续的方式在体内递送这些因子的脂质体制剂。在最终的目标中，评价用于原位再活化Tem和用于诱导局部和全身抗肿瘤反应的每个因子的治疗功效。后者将使用已建立的异种移植模型来完成，在该模型中，将未破裂的人肿瘤块手术植入SCID或NOD-SCID/IL 2受体中。链无效小鼠。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保持存活并且可预测地对细胞因子信号响应延长的时间段。在将加载因子的脂质体接种到异种移植物中之后，监测肿瘤杀伤和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策略奠定基础，这些策略可用于增强我们目前癌症疫苗临床试验的有效性。 存在于癌症患者肿瘤内和周围的记忆T细胞具有识别和杀死肿瘤细胞的潜力，但它们通常无法控制这些患者中癌细胞的生长和扩散。这种故障部分是由于存储器单元的非响应或静止状态。我们正在研究这种静止状态的原因，设计逆转这种无反应性的策略，并测试重新激活肿瘤相关记忆T细胞的治疗益处。