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.

项目摘要 理解和操纵细胞信号传导过程对于收养细胞疗法（ACT）至关重要， 在治疗癌症和糖尿病等疾病方面表现出巨大的希望。当前许多挑战 制造这些治疗剂与我们缺乏对离体T细胞激活的控制有关。尽管 在理解细胞外信号线索如何影响细胞内方面已经取得了巨大进步 州，我们对管理这些过程的详细机制的理解是不完整的。安装 证据表明，细胞信号传导受信号结构的物理排列调节 细胞表面。但是，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级尺寸，这非常困难，该结构低于传统的分辨率限制 光学显微镜。 这项研究将提供至关重要的信息，以阐明空间组织在T细胞中的作用 调节以及测试新型工具研究和操纵纳米级结构的可行性。 我们的目标是确定信号分子的3D空间排列如何影响T细胞行为。做 因此，我们将使用DNA折纸纳米结构将配体排列到纳米级3D模式中，然后将其呈现 对T细胞的图案化配体并表征信号传导动力学，我们还将评估相关参数 例如T细胞增殖率和IL-2分泌。我们的理由是，通过定义 配体排列和T细胞信号传导，我们将更好地了解信号分子的组织如何 在细胞表面调节细胞内途径，这将指导开发优化试剂 有效的离体T细胞激活。该项目将利用纳米技术，生物化学和细胞来 完成三个具体目标：1）确定细胞外受体激酶动力学之间的关系 和3D刺激配体排列，2）确定抑制受体对T细胞的空间依赖性 激活和3）创建模式的T细胞信号试剂，该试剂可能触发离体原发性T细胞激活。 我们将定义信号分子的空间组织与细胞内之间的关系 途径，并在建立纳米图案的免疫疗法试剂基础上。这些知识会 让我们更深入地理解T细胞激活和分化的基础机制，从而实现 用于癌症，糖尿病和其他疾病的细胞疗法的有效生产。