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

• 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
• 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

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.

项目总结