Defining the role of ligand spatial organization in T cell signaling with DNA origami

用 DNA 折纸定义配体空间组织在 T 细胞信号传导中的作用

基本信息

批准号：
10680089
负责人：
Konlin Shen
金额：
$ 6.95万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-06-01 至 2024-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10680089
关键词：
3-Dimensional Address Adoptive Cell Transfers Affect Antibodies Antigen Presentation Antigen-Presenting Cells Autoimmune Diseases Binding Biochemistry Biological Process Cell Signaling Process Cell Therapy Cell model Cell surface Cells Cellular biology Cues DNA Dependence Development Diabetes Mellitus Disease Engineering Exclusion Foundations Goals Height Human Immune signaling Immunotherapy Interleukin-2 Knowledge Lateral Learning Ligands Linker DNA Lupus Malignant Neoplasms Mediating Methods Nanotechnology Nucleotides Pathway interactions Patients Pattern Peptide/MHC Complex Phosphotransferases Positioning Attribute Process Production Public Health Reagent Research Resolution Resources Role Signal Pathway Signal Transduction Signaling Molecule Single-Stranded DNA Structure Surface T cell regulation T cell therapy T-Cell Activation T-Cell Proliferation T-Cell Receptor T-Lymphocyte Techniques Therapeutic Work cancer immunotherapy cancer therapy cell behavior design electron beam lithography engineered T cells extracellular feasibility testing immunological synapse immunological synapse formation innovation insight interest light microscopy manufacture migration nanometer resolution nanopattern nanoscale novel programmed cell death protein 1 receptor scaffold stoichiometry success tool

项目摘要

Project Summary Understanding and manipulating cell signaling processes is crucial for adoptive cell therapies (ACT), which show significant promise in treating diseases such as cancer and diabetes. Many of the current challenges in manufacturing these therapeutics are related to our lack of control over ex vivo T cell activation. Though tremendous progress has been made in understanding how extracellular signaling cues influence intracellular states, our understanding of detailed mechanisms governing these processes is incomplete. Mounting evidence suggests that cell signaling is regulated by the physical arrangement of signaling structures at the surface of cells. However, determining how the spatial arrangement of signaling structures guides cell behavior is very difficult due to the nanoscale size of these structures, which is below the resolution limit of traditional light microscopy. This study will provide crucial information towards elucidating the role of spatial organization in T cell regulation, as well as test the feasibility of novel tools I have designed to study and manipulate structures on the nanoscale. The objective of this study is to determine how 3D spatial arrangements of signaling molecules affect T cell behavior. To do this, I will arrange ligands into nanoscale 3D patterns, then present these patterned ligands to T cells and characterize signaling dynamics, as well as T cell manufacturing parameters such as T cell proliferation rate and IL-2 secretion. The rationale for this work is that by defining the relationship between ligand arrangement and T cell signaling, we will better understand how the organization of signaling molecules at the cell surface regulates intracellular pathways, which will guide the development of optimized reagents for efficient ex vivo T cell activation. I will accomplish this goal using an integrated approach drawing from nanotechnology, biochemistry, and cell biology. This project will conduct three Specific Aims: 1) determine the relationship between extracellular receptor kinase dynamics and 3D stimulatory ligand arrangement, 2) determine the spatial dependence of inhibitory receptors on T cell activation, and 3) create patterned T cell signaling reagents that can trigger ex vivo primary T cell activation. My project is highly innovative because it will use DNA origami as a method to pattern ligands with single nanometer-resolution, a degree of precision rivaled only by advanced cleanroom techniques such as electron beam lithography. This project’s significance lies in defining the relationship between the spatial organization of signaling molecules and intracellular pathways, and in establishing the foundation for nanopatterned immunotherapy reagents. This knowledge will allow us to more deeply understand the mechanisms underlying T cell activation and differentiation, enabling efficient and efficacious manufacturing of cell therapies for cancer, diabetes, and other diseases.

项目概要了解和操纵细胞信号传导过程对于过继细胞疗法 (ACT) 至关重要，该疗法在治疗癌症和糖尿病等疾病方面显示出巨大的前景。制造这些疗法与我们缺乏对离体 T 细胞激活的控制有关。在理解细胞外信号传导如何影响细胞内信号方面取得了巨大进展表明，我们对控制这些过程的详细机制的理解并不完整。表明细胞信号传导受到细胞表面信号传导结构的物理排列的调节然而，确定信号结构的空间排列如何指导细胞行为是非常重要的。由于这些结构的纳米级尺寸，低于传统光的分辨率极限，因此很困难显微镜。这项研究将为阐明空间组织在 T 细胞调节中的作用提供重要信息，以及测试我设计的用于研究和操纵纳米尺度结构的新工具的可行性。本研究的目的是确定信号分子的 3D 空间排列如何影响 T 细胞为此，我会将配体排列成纳米级 3D 图案，然后将这些图案化的配体呈现给 T 细胞和表征信号动力学，以及 T 细胞制造参数，例如 T 细胞这项工作的基本原理是通过定义配体之间的关系。排列和 T 细胞信号传导，我们将更好地了解信号分子在细胞表面调节细胞内途径，这将指导优化试剂的开发，以实现高效我将使用纳米技术的综合方法来实现这一目标，该项目将进行三个具体目标：1）确定关系。细胞外受体激酶动力学和 3D 刺激配体排列之间，2) 确定空间抑制性受体对 T 细胞激活的依赖性，以及 3) 创建图案化 T 细胞信号转导试剂可以触发体外初级 T 细胞激活我的项目具有高度创新性，因为它将使用 DNA 折纸作为方法以单纳米分辨率对配体进行图案化，其精确度只有先进的技术才能匹敌该项目的意义在于定义了洁净室技术，例如电子束光刻。信号分子的空间组织与细胞内通路之间的关系，以及这些知识将为纳米图案免疫治疗试剂奠定基础。深入了解T细胞激活和分化的机制，实现高效、高效生产癌症、糖尿病和其他疾病的细胞疗法。