Novel mechanisms regulating PD-1 signaling and function

调节 PD-1 信号传导和功能的新机制

基本信息

批准号：
9158876
负责人：
Adam Mor
金额：
$ 42.38万
依托单位：
NEW YORK UNIVERSITY SCHOOL OF MEDICINE
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-20 至 2021-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9158876
关键词：
Adaptor Signaling Protein Adhesions Affect Antibodies Apoptosis Regulation Gene Binding Binding Sites Biochemical Biological Assay Biological Markers Blocking Antibodies Cell Adhesion Cell Survival Cessation of life Clinical Clinical Trials Co-Immunoprecipitations Collaborations Cytoplasmic Tail Development Disease remission Event Fluorescence Microscopy Fluorescence Resonance Energy Transfer Genes Goals Human Immune Immune response Immune system Infection Leukocytes Ligand Binding Ligation Lymphocyte Lymphoproliferative Disorders Malignant Neoplasms Maps Mass Spectrum Analysis Measures Mediating Microscopy Molecular Monitor Mutate Mutation PTPN11 gene Pathway interactions Patients Pharmaceutical Preparations Phosphotransferases Play Proteins Public Health RNA Interference Regulation Resolution Role Signal Pathway Signal Transduction Site-Directed Mutagenesis System T-Lymphocyte Tail Testing Therapeutic Therapeutic Agents Work X-Linked lymphoproliferative disorders base cancer cell cancer immunotherapy cancer therapy checkpoint therapy cytokine design drug discovery fighting immunological synapse improved in vivo insight knock-down mouse model neoplastic cell novel novel strategies novel therapeutics overexpression potential biomarker programs promoter receptor release of sequestered calcium ion into cytoplasm research study response tumor tumor progression

项目摘要

ABSTRACT Immune checkpoint therapies block inhibitory receptors on T cells in efforts to augment anti-tumor immune responses. The Programmed death-1 (PD-1) pathway is a critical inhibitory checkpoint in T cells and antibodies blocking PD-1 promote immune-mediated identification and clearance of malignancies. Cancer immunotherapies, like anti-PD-1 antibodies, represent a paradigm shift in cancer treatment because they do not target the tumor cell directly. Instead, by harnessing the immune system, anti-PD-1 therapies can elicit durable clinical responses in multiple tumor types, including long-term remissions. Unfortunately, the benefit of anti-PD-1 therapy is realized in only a fraction of patients. As such, developing additional therapeutics that can better inhibit this pathway will extend the benefit of this approach to many additional patients. To achieve this goal, a deeper insight into the signaling events downstream of PD-1 and characterization of the molecular mechanism(s) by which PD-1 exerts its inhibitory effect is critical. Toward this end, we have taken an unbiased approach to identify novel targets in the PD-1 checkpoint pathway. We used advanced mass spectrometry to identify new signaling pathways downstream of PD-1. Using this system, we identified multiple candidates that we categorized into two groups: promoters or suppressors of PD-1 functions. Excitingly, we found that silencing PD-1 promoters, and not suppressors, with RNAi abrogated the inhibitory effects of PD-1. We now seek to elucidate the mechanisms by which these proteins affect PD-1 function and determine their suitability as either targets for drug discovery or biomarkers for PD-1 treatment with three specific aims: In the first aim we will confirm that the interaction between PD-1 and the newly discovered promoters is direct in primary human T cells. In order to study the physical interactions, we will utilize high-resolution microscopy. We will incorporate an in vivo mouse model of cancer into our studies, with the goal of establishing a role for our candidates in tumor progression/regression. In the second aim we will further characterize the functional role of PD-1 suppressors in primary human T cells and measure cytokine secretion, proliferation, and cellular adhesion in response to stimulation. In the third aim we will investigate the subcellular distribution of PD-1 modulators and measure their ability to form mature immunological synapses. Taken together, the execution of these aims will help define a mechanistic understanding of PD-1 function to guide the development of improved cancer immunotherapies and potential biomarkers of anti-PD-1 responses.

抽象的免疫检查点疗法阻止T细胞上的抑制性受体，以增强抗肿瘤免疫回答。编程的死亡1（PD-1）途径是T细胞和阻断PD-1的抗体促进了免疫介导的鉴定和恶性肿瘤的清除率。癌症免疫疗法，例如抗PD-1抗体，代表癌症治疗的范式转移不直接靶向肿瘤细胞。相反，通过利用免疫系统，抗PD-1疗法可以引起多种肿瘤类型的持久临床反应，包括长期减免。不幸的是，抗PD-1治疗仅在一小部分患者中实现。因此，开发其他可以更好地抑制这种途径将向许多其他患者扩展这种方法的好处。实现这一目标目标，对PD-1下游的信号事件的深入了解和分子的表征 PD-1发挥其抑制作用的机制至关重要。为此，我们采取了公正的态度在PD-1检查点途径中识别新目标的方法。我们使用高级质谱法确定PD-1下游的新信号通路。使用此系统，我们确定了多个候选人我们分为两组：PD-1功能的启动子或抑制器。令人兴奋的是，我们发现将PD-1启动子静音，而不是抑制子，RNAi消除了PD-1的抑制作用。我们现在寻求阐明这些蛋白质影响PD-1功能的机制并确定其适用性作为药物发现的靶标，或以三个特定目的进行PD-1处理的生物标志物：在第一个目标中我们将确认PD-1与新发现的启动子之间的相互作用是直接的人类T细胞。为了研究物理相互作用，我们将利用高分辨率显微镜。我们将将癌症的体内小鼠模型纳入我们的研究，目的是确立我们的作用肿瘤进展/回归中的候选者。在第二个目标中，我们将进一步描述原代人T细胞中的PD-1抑制剂，并测量细胞因子分泌，增殖和细胞响应刺激的粘附。在第三个目标中，我们将研究PD-1的亚细胞分布调节器并衡量其形成成熟免疫突触的能力。两者一起执行这些目标将有助于定义对PD-1功能的机械理解，以指导开发改善了抗PD-1反应的癌症免疫疗法和潜在的生物标志物。