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.

肿瘤相关的CD4+和CD8+ T细胞具有效应子记忆表型（TEM） 人类非小细胞肺肿瘤和卵巢癌的微环境，但无法控制肿瘤 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 他们未能通过T细胞受体（TCR）响应激活信号。通过阐明被捕地 在TCR途径中，通过定义启动此逮捕的分子和细胞事件，应该是 可以设计和测试策略以重新激活原位。 TEM内的局部重新激活 治疗的肿瘤微环境有望导致（a）T细胞介导的原位杀死肿瘤细胞的杀死，（b） 将肿瘤抗原释放到循环中，（c）全身抗肿瘤免疫的产生，（d）T细胞 识别和消除与最初治疗或远离或远处的现有肿瘤的识别和消除 肿瘤部位。结合了多光谱载体流式细胞仪，共聚焦显微镜，蛋白质印迹 RTPCR我们的第一个目的是确定在TCR途径中的何处转导信号被阻塞。在 相关的第二个目标确定与触发TCR停滞有因果关系的细胞和分子 通过细胞耗竭并添加协议，并通过监视选定分子对启动的影响 此调节信号检查点。我们的初步研究已经定位了信号检查点的位置 发生在PLC-γ上游的某个地方，TCR信号停滞已与TGF-β1有因果关系 证明我们实验方案的可行性和生存能力。从这些结果获得的结果 机械研究将用于目标2和3中，以识别直接起作用或 间接在T细胞上扭转其无反应性，并开发和测试脂质体配方 旨在在体内以当地和持续的方式提供这些因素。在最终目标中 每个因素重新激活TEM原位并诱导局部和全身抗肿瘤的治疗功效 评估响应。后者将使用已建立的异种移植模型来实现，其中无干扰 将人肿瘤的碎片手术植入SCID或NOD-SCID/IL2受体γ链虫小鼠中。 在这些异种移植物中，保留了肿瘤微环境，并且与肿瘤相关的白细胞保留 长时间的可行且可预测的对细胞因子信号的反应。接种后接种 将因子脂质体加载到异种移植物肿瘤杀死和TEM反应模式中以确定 每种脂质体制剂的治疗功效。这些研究有望为 设计可用于增强我们当前癌症疫苗临床试验功效的策略。