Contribution of PAG to Immune Synapse Organization and PD-1 Function

PAG 对免疫突触组织和 PD-1 功能的贡献

基本信息

批准号：
10538164
负责人：
Emily Kathryn Moore
金额：
$ 5.18万
依托单位：
COLUMBIA UNIVERSITY HEALTH SCIENCES
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-07-15 至 2026-07-14
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10538164
关键词：
Actins Adhesions Adoptive Cell Transfers Adverse effects Anatomy Antibodies Antibody Therapy Antigen-Presenting Cells Antitumor Response Architecture Autoimmune Diseases Automobile Driving Binding Biological Assay C-terminal CD28 gene Cause of Death Cell Line Cell membrane Cell physiology Cells Clinical Complex Confocal Microscopy Cytoskeleton Enzyme-Linked Immunosorbent Assay Enzymes Event Fellowship Flow Cytometry Fluorescence Resonance Energy Transfer Fostering Future Glycosphingolipids Goals Human Imaging Techniques Immune Immune Targeting Immune checkpoint inhibitor Immune response Immunology Immunotherapy Integral Membrane Protein Knock-out Ligand Binding Ligation Link Lipids MAP Kinase Gene Malignant Neoplasms Measures Mediating Medicine Membrane Membrane Microdomains Mentorship Methods Mind Modeling Mus Mutate Palmitic Acylation Site Pathway interactions Patients Pattern Peripheral Pharmacology Phosphoproteins Phosphorylation Physicians Process Production Proteins Regimen Research Research Personnel Resources Role Shapes Signal Pathway Signal Transduction Signaling Protein Structure Synapses T-Cell Activation T-Cell Receptor T-Lymphocyte Technology Testing Therapeutic Therapeutic antibodies Total Internal Reflection Fluorescent Tumor-infiltrating immune cells Tyrosine Phosphorylation Site Work anti-PD-1 anti-PD1 antibodies antibody immunotherapy cancer cell cancer immunotherapy cancer therapy career cytokine cytotoxicity immune function immune-related adverse events immunological synapse improved in vivo innovation interest mortality new therapeutic target novel novel therapeutics polymerization prevent programmed cell death protein 1 protein protein interaction recruit response skill acquisition skills synaptic function synaptogenesis targeted cancer therapy targeted treatment therapeutic target tumor

项目摘要

PROJECT SUMMARY/ABSTRACT Cancer remains the second leading cause of death in the US. Immunotherapy seeks to bolster immune cells’ ability to target malignant cells and has brought immense improvements in the field. One important inhibitory protein in T cells, Programmed Cell Death Protein 1 (PD-1), has become an invaluable target for cancer immunotherapy. While anti-PD-1 antibody therapy is extremely successful in some patients, in many others, it fails to help or causes complications, including cancer hyper-progression and immune-related adverse events. Study of the inhibitory transmembrane protein Phosphoprotein Associated with Glycosphingolipid Rich Microdomains 1 (PAG), a downstream target of PD-1 signaling, will help us better understand the PD-1 pathway, and offer another, perhaps more nuanced, target to potentially improve response rates and/or avoid immune- related adverse events. As a link between lipid-rich/signaling-protein-rich membrane regions and the actin cytoskeleton, PAG is an exciting and novel target for manipulating immune function. Prior therapeutic methods of immune manipulation all disrupt ligand binding or enzyme function. In contrast, innovative use of an anti-PAG antibody to simply disrupt appropriate PAG localization within the synapse could disturb immune synapse architecture. Synapse organization is tightly regulated to prevent inappropriate immune responses, but the precise interaction between cytoskeletal dynamics and synaptic organization is not fully understood. Investigating the role of PAG in this process could provide added clarity. To test the hypothesis that PAG works with actin to control T cell synapse organization and facilitate T cell receptor (TCR) and PD-1 signaling, PAG will be studied in a human T cell line and a murine PAG-knockout model. Methods will include confocal and TIRF microscopy, FRET and proximity ligation assay, flow cytometry, ELISA, adhesion and cytotoxicity assays, and murine tumor models. To understand the contribution of the actin binding domain of PAG on its localization, protein-protein interactions, and T cell activation and effector functions, the primary hypothesis will be examined through the following specific aims: Aim 1. Define the role of the PAG-actin link in driving T cell synapse architecture and stability. Aim 2. Determine the impact of PAG-actin interactions on T cell functions downstream of TCR and PD-1 signaling. Aim 1 will demonstrate the role of PAG in T cells synapse anatomy, and the physical impacts of a PAG-targeting therapy. Aim 2 will illuminate which PD-1 downstream targets are dependent on the PAG-actin link, and provide evidence for whether PAG and PD-1 could serve as good co-targets in cancer therapy regimens. Ultimately, this study will illuminate crucial control mechanisms associated with T cell synapse organization, opening more avenues of targeting the immune synapse. Furthermore, the mentorship and resources in Dr. Adam Mor’s lab and the Columbia MSTP, will foster invaluable technical and professional skill development for a career in medicine and immunology research.

项目概要/摘要癌症仍然是美国第二大死亡原因，免疫疗法旨在增强免疫细胞的功能。靶向恶性细胞的能力给抑制领域带来了巨大的进步。 T 细胞中的蛋白质，程序性细胞死亡蛋白 1 (PD-1)，已成为癌症的宝贵靶点虽然抗 PD-1 抗体疗法在某些患者中非常成功，但在其他许多患者中，却效果不佳。无法帮助或导致并发症，包括癌症过度进展和免疫相关的不良事件。与富含鞘糖脂相关的抑制性跨膜蛋白磷酸蛋白的研究微结构域 1 (PAG) 作为 PD-1 信号传导的下游靶点，将帮助我们更好地了解 PD-1 通路，并提供另一个可能更微妙的目标，以潜在地提高反应率和/或避免免疫- 作为富含脂质/富含信号蛋白的膜区域和肌动蛋白之间的联系。细胞骨架，PAG 是操纵免疫功能的令人兴奋的新颖靶点。免疫操作的所有破坏配体结合或酶功能相反，抗PAG的创新使用。简单地破坏突触内适当的 PAG 定位的抗体可能会干扰免疫突触突触组织受到严格监管以防止不适当的免疫反应，但是细胞骨架动力学和突触组织之间的精确相互作用尚未完全了解。研究 PAG 在此过程中的作用可以进一步澄清 PAG 的假设。与肌动蛋白一起控制 T 细胞突触组织并促进 T 细胞受体 (TCR) 和 PD-1 信号传导方面，PAG 将在人类 T 细胞系和小鼠 PAG 敲除模型中进行研究，方法将包括。共聚焦和 TIRF 显微镜、FRET 和邻近连接测定、流式细胞术、ELISA、粘附和细胞毒性测定和小鼠肿瘤模型了解肌动蛋白结合域的贡献。 PAG 的定位、蛋白质-蛋白质相互作用以及 T 细胞激活和效应功能，这是主要的将通过以下具体目标来检验假设：目标 1. 定义 PAG-肌动蛋白链接的作用目标 2. 确定 PAG-肌动蛋白相互作用的影响。目标 1 将展示 PAG 在 T 细胞中的作用。突触解剖结构，以及 PAG 靶向治疗的物理影响，目标 2 将阐明哪种 PD-1。下游靶标依赖于 PAG-肌动蛋白链接，并为 PAG 和 PD-1 是否存在提供证据最终，这项研究将阐明关键的控制。与 T 细胞突触组织相关的机制，开辟更多针对免疫系统的途径此外，Adam Mor 博士的实验室和哥伦比亚 MSTP 的指导和资源将促进突触的发展。为医学和免疫学研究职业提供宝贵的技术和专业技能发展。