Probing mesoscale receptor organization in T cell signaling with DNA origami

用 DNA 折纸探测 T 细胞信号传导中的中尺度受体组织

• 批准号: 10726455
• 负责人: Shawn M Douglas
• 金额: $ 20.19万
• 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: 2023-05-19 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10726455
• 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; Cues; DNA; Dependence; Development; Diabetes Mellitus; Disease; Engineering; Exclusion; Foundations; Goals; Health Benefit; Height; Human; Immune signaling; Immunotherapy; Interleukin-2; Knowledge; Lateral; Learning; Ligands; Linker DNA; Malignant Neoplasms; Mediating; Nanostructures; Nanotechnology; Nucleotides; Pathway interactions; Patients; Pattern; Peptide/MHC Complex; Phosphotransferases; Positioning Attribute; Process; Production; Public Health; Reagent; Resolution; 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; Therapeutic; Work; cancer immunotherapy; cancer therapy; cell behavior; engineered T cells; extracellular; feasibility testing; immunological synapse; immunological synapse formation; insight; interest; light microscopy; manufacture; migration; nanopattern; nanoscale; novel; programmed cell death protein 1; receptor; scaffold; stoichiometry; success; tool; two-dimensional

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 to study and manipulate structures on the nanoscale. Our objective is to determine how 3D spatial arrangements of signaling molecules affect T cell behavior. To do so, we will use DNA origami nanostructures to arrange ligands into nanoscale 3D patterns, then present these patterned ligands to T cells and characterize signaling dynamics, we will also assess ACT-relevant parameters such as T cell proliferation rate and IL-2 secretion. Our rationale 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. This project will leverage nanotechnology, biochemistry, and cell to accomplish 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. We will define 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 了解和操纵细胞信号传递过程对于过继细胞疗法(ACT)至关重要，ACT 在治疗癌症和糖尿病等疾病方面显示出巨大的希望。中国当前面临的许多挑战 制造这些治疗药物与我们缺乏对体外T细胞激活的控制有关。尽管 在理解细胞外信号信号如何影响细胞内方面已经取得了巨大的进展 但是，我们对管理这些进程的详细机制的理解是不完整的。安装 有证据表明，细胞信号是由信号结构的物理排列来调节的。 细胞表面。然而，确定信号结构的空间排列如何指导细胞行为 由于这些结构的纳米尺寸，这是非常困难的，这低于传统的分辨率极限 光学显微镜。 这项研究将为阐明T细胞中空间组织的作用提供关键信息 监管，以及测试新工具在纳米尺度上研究和操纵结构的可行性。 我们的目标是确定信号分子的3D空间排列如何影响T细胞的行为。去做 因此，我们将使用DNA折纸纳米结构将配体排列成纳米级的3D图案，然后呈现这些 我们还将评估与ACT相关的参数 如T细胞增殖率和IL-2分泌。我们的理论基础是，通过定义 配体的排列和T细胞的信号传递，我们会更好地了解信号分子是如何组织的 在细胞表面调节细胞内途径，这将指导优化试剂的开发 高效的体外T细胞激活。该项目将利用纳米技术、生物化学和细胞来 完成三个特定目标：1)确定胞外受体激酶动力学之间的关系 和3D刺激配体排列，2)决定抑制受体对T细胞的空间依赖性 激活，以及3)创造图案化的T细胞信号试剂，可以触发体外初级T细胞激活。 我们将定义信号分子的空间组织和细胞内的关系 途径，并建立纳米免疫治疗试剂的基础。这一知识将会 使我们能够更深入地了解T细胞激活和分化的机制，使 高效生产治疗癌症、糖尿病和其他疾病的细胞疗法。