BME: DNA Origami for Investigating and Reprogramming Cell Signaling

PI：LaBean，Thom提案编号：1603179细胞信号通路的复杂网络组成了细胞间的通讯网络，负责从体内平衡(维持身体内的平衡)到伤口愈合、发育和免疫的一切。这些通讯系统利用相邻细胞之间的直接接触，通过特定的纳米级组织和细胞表面受体和配体的呈现。该项目将使用一种名为DNA折纸(或DNA折叠)的独特分子工程工具，来确定控制细胞外膜蛋白质(主要组织相容性复合体或MHC)的产生的基因如何与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的重要生物医学问题。因此，这项建议中的工作有可能使一个强大的工具包，以执行受体动力学的基础生物学研究和开发治疗免疫疾病。这个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、定向和呈现生物活性分子给活细胞，以便了解、促进、中断和重新编程细胞信号中涉及的分子相互作用。细胞信号通路的复杂网络组成了细胞间的通讯网络，负责从动态平衡到伤口愈合、到发育和免疫的一切。这些通讯系统通常利用相邻细胞之间的直接接触，通过特定的纳米级组织和呈现与它们相互作用的细胞表面受体和配体。具体地说，该项目将研究主要组织相容性复合体(MHC)的呈现间隔和多样性，以及它与活细胞表面的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的第一步，T细胞激活影响正常免疫功能、癌症和自身免疫性疾病的重要生物医学问题。DNA折纸是DNA纳米技术的一个子集，它将与设计的架构、图案和结构加固一起组装，以测试细胞信号科学中的一系列假设，这些假设将很难或不可能通过其他编程较少的实验方法进行测试。现在可以追求新的基础科学，以及使用显示蛋白质、配体、受体、适配子、小分子和其他细胞效应器的DNA折纸分子组件来测试可能的治疗策略。用于探测单个细胞间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，代表了一个在理解生物学和影响人类健康方面具有重大长期潜力的新的智力前沿。该计划将导致自然生物兼容分子组织器的开发，用于各种生物和生物医学应用。这项研究的结果可能会为理解T细胞激活和细胞间通讯中的重要事件提供重大进展。该项目未来的扩展可能涉及将这里开发的分子材料和方法应用于其他对理解和改善人类健康至关重要的细胞信号通路。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，并将在这一项目上加倍努力

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