BME: DNA Origami for Investigating and Reprogramming Cell Signaling

PI：LaBean，Thom 提案编号：1603179 复杂的细胞信号通路网络构成了细胞间的通信网络，负责从稳态（维持体内平衡）到伤口愈合、发育和免疫的一切。这些通信系统通过特定的纳米级组织以及细胞表面受体和配体的呈现，利用相邻细胞之间的直接接触。该项目将使用一种独特的分子工程工具，称为 DNA 折纸（或 DNA 折叠），来确定控制细胞外膜上蛋白质（主要组织相容性复合物或 MHC）产生的基因如何与 T 细胞受体 (TCR) 相互作用。 MHC 和 TCR 的相互作用是 T 细胞激活过程的第一步，它影响正常免疫功能、癌症和自身免疫性疾病中的重要生物医学问题。 因此，该提案中的工作有可能成为一个强大的工具包，用于对受体动力学进行基础生物学研究并开发免疫性疾病的疗法。 该项目的目标是利用结构 DNA 纳米技术提供的丰富的分子工程工具来专门组织、定向和向活细胞呈现生物活性分子，以便理解、促进、中断和重新编程细胞信号传导中涉及的分子相互作用。 细胞信号通路的复杂网络构成了细胞间的通信网络，负责从稳态到伤口愈合、发育和免疫等一切。这些通信系统通常通过特定的纳米级组织以及与其相互作用的细胞表面受体和配体的呈现，利用相邻细胞之间的直接接触。 具体来说，该项目将检查主要组织相容性复合体 (MHC) 的间距和呈现多样性及其与活细胞表面 T 细胞受体 (TCR) 的相互作用。 MHC 和 TCR 的相互作用是涉及 T 细胞激活的复杂生化级联反应的第一步，该级联反应影响正常免疫功能、癌症和自身免疫性疾病中的重要生物医学问题。 DNA折纸是DNA纳米技术的一个子集，它将与设计好的架构、图案和结构加固组装在一起，以测试细胞信号传导科学中的一系列假设，而这些假设很难或不可能通过其他可编程性较差的实验方法进行测试。 现在可以探索新的基础科学，并使用展示蛋白质、配体、受体、适体、小分子和其他细胞效应物的 DNA 折纸分子组装体测试可能的治疗策略。用于探测单个细胞与细胞相互作用的精心设计的实验装置与基于 DNA 的纳米结构的可编程分子识别平台相结合，代表了一个新的知识前沿，在理解生物学和影响人类健康方面具有巨大的长期潜力。该计划将导致开发用于各种生物和生物医学应用的天然生物相容性分子组织体。这项研究的结果可能为理解 T 细胞激活和细胞间通讯的重要事件提供重大进展。该项目的未来扩展可能涉及将此处开发的分子材料和方法应用于对理解和改善人类健康至关重要的其他细胞信号传导途径。研究生将获得宝贵的教育和研究培训机会。 PI 一直让本科生、高中生和代表性不足的学生参与研究项目，并将在该项目上加倍努力

PI: LaBean, Thom Proposal Number: 1603179Complex webs of cell signaling pathways make up cell-to-cell communication networks responsible for everything from homeostasis (maintaining equilibrium within our body), to wound healing, to development, and to immunity. These communication systems make use of direct contact between neighboring cells through specific nanometer-scale organization and presentation of cell surface receptors and ligands. This project will use a unique molecular engineering tool, termed DNA origami (or DNA folding), to determine how genes that control the production of proteins on the outer membranes of cells (major histocompatibility complex or MHC) interact with T-cell receptors (TCR). The interaction of the MHC and the TCR is the first step in the T-cell activation process, which impacts important biomedical issues in normal immune function, cancer, and autoimmune disease. Thus, the work in this proposal has the potential to enable a powerful toolkit for performing fundamental biological studies on receptor dynamics and developing therapies for immunological disorders. The goal of this project is to employ the rich palette of molecular engineering tools provided by structural DNA nanotechnology to specifically organize, orient, and present biologically active molecules to living cells in order to understand, facilitate, interrupt, and reprogram molecular interactions involved in cell signaling. Complex webs of cell signaling pathways make up the cell-to-cell communication networks responsible for everything from homeostasis, to wound healing, to development, and to immunity. These communication systems often make use of direct contact between neighboring cells via specific nanometer-scale organization and presentation of cell surface receptors and ligands they interact with. Specifically, the project will examine the spacing and multiplicity of presentation of the major histocompatibility complex (MHC) and its interaction with T-cell receptors (TCR) on the surfaces of living cells. The interaction of MHC and TCR is the first step in the complex biochemical cascade involving T-cell activation that impacts important biomedical issues in normal immune function, cancer, and autoimmune disease. DNA origami, a subset of DNA nanotechnology, will be assembled with designed architectures, patterns, and structural reinforcement in order to test a range of hypotheses in cell signaling science that would be very difficult or impossible to test by other, less programmable experimental methods. Novel basic science can now be pursued as well as testing of possible therapeutic strategies using DNA origami molecular assemblies displaying proteins, ligands, receptors, aptamers, small molecules, and other cell effectors. The combination of well-engineered experimental set-ups for probing individual cell-cell interactions allied with the programmable molecular recognition platforms of DNA-based nanostructures represents a new intellectual frontier with significant long-term potential in understanding biology and affecting human health. The program will lead to development of naturally biocompatible molecular organizers for a variety of biological and biomedical applications. Results from this study may provide major advances in understanding important events in T-cell activation and in cell-to-cell communications in general. Future extensions of this project could involve application of molecular materials and methods developed here toward other cell signaling pathways critical to understanding and improving human health. Graduate students will gain valuable opportunities for education and research training. The PI has consistently involved undergraduate, high school, and under-represented students in research programs and will redouble efforts to do so on this project