Spatially and Temporarily Resolved Precision Delivery for Quantitative Biological Studies

用于定量生物学研究的空间和暂时解决的精确传递

• 批准号: 10799275
• 负责人: Hang Ren
• 金额: $ 14.55万
• 依托单位: UNIVERSITY OF TEXAS AT AUSTIN
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-08-16 至 2027-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10799275
• 关键词: Biological; Biological Process; Blood coagulation; Cells; Chemicals; Complex; Disease; Electrodes; Environment; Generations; Health; Heterogeneity; Human; Imaging Techniques; Immune response; In Situ; Inflammatory Response; Investigation; Ions; Location; Maps; Methods; Modality; Neurons; Nitric Oxide; Physiologic pulse; Physiological; Physiological Processes; Proteins; Reactive Nitrogen Species; Reactive Oxygen Species; Research; Spatial Distribution; Stimulus; System; Vesicle; analytical tool; digital delivery; interest; nano; nanoparticle; nanoscale; neurotransmission; response; single cell analysis; temporal measurement; tool; uptake

Project Summary Spatial and temporal heterogeneity in the cellular environment has profound implications in biological processes related to human health/disease. Many single-cell analytical tools have been developed over the years to reveal the heterogeneity among cells, e.g., the spatial distribution of chemicals and ions. However, one missing piece in the single-cell analysis is the ability to reveal quantitatively the spatial and temporal heterogeneity cellular response to chemical stimulus. This is challenging because controlling the exact concentration of chemicals at a specific location depends on the interplay between dynamics of mass transport in the complex cellular environment and the reactivity of the molecules. Indeed, some physiologically important molecules, including reactive oxygen species (ROS), reactive nitrogen species (RNS), are highly reactive and have short lifetimes. A tool for precision delivery of molecules, including these reactive ones, are necessary to quantitatively study their effect at the single-cell level. Our research lab will focus on developing nanoscale precision delivery tools to quantitatively control the delivery of molecules of biological interest, including those highly reactive ones. The strategy is based on a functionalized nanopipette electrode that is capable of in situ generation of the molecule of interest electrochemically with spatial and temporal control. This will be demonstrated by the delivery of nitric oxide (NO), a reactive molecule whose transient concentration is important in neuron transmission, immune response, and blood coagulation. Spatial and temporally resolved delivery is achieved by combining the electrochemical chemical delivery system with nanoscale electrochemical imaging techniques. This delivery modality can be extended to other reactive molecules, including H2S, CO, and ROS. In addition, we will develop a precision delivery tool called digital delivery, where we will precisely control the number of biomolecules or other non- biological entities being delivered, including proteins and nanoparticles, by counting their number during the delivery in a resistive pulse fashion. Lastly, we will quantitatively map the spatially resolved rate of uptake of the molecules being delivered. Ultimately, the precision delivery methods developed in our proposed research will enable quantitative investigation of many fundamental biological and physiological questions related to the reactive molecules at the single-cell level. For example, the tools can be used to reveal the spatial and temporal heterogeneity in the neuron response by precision delivery of neuron transmitters or their vesicles. The modality can also be applied to quantitatively modulate or stimulate the inflammatory response at the single-cell level.

项目摘要 细胞环境中的空间和时间异质性在生物过程中具有深远的影响 与人类健康/疾病有关。多年来，已经开发了许多单细胞分析工具来揭示 细胞之间的异质性，例如化学物质和离子的空间分布。但是，一件丢失的作品 在单细胞分析中，可以定量揭示空间和时间异质性细胞的能力 对化学刺激的反应。这是具有挑战性的，因为控制化学物质的确切浓度 特定位置取决于复杂细胞中质量传输动力学之间的相互作用 环境和分子的反应性。实际上，某些具有生理上重要的分子，包括 活性氧（ROS），反应性氮（RN）具有高反应性，寿命很短。一个 精确递送分子的工具，包括这些反应性的工具，对于定量研究它们是必要的 在单细胞水平上的效果。 我们的研究实验室将着重于开发纳米​​级精确输送工具，以定量控制 传递生物学兴趣的分子，包括那些高反应性的分子。该策略基于 功能化的纳米纤维电极，能够原位产生感兴趣的分子 通过空间和时间控制进行电化学。这将通过一氧化氮（NO）的输送来证明这一点 一种反应性分子，其瞬时浓度在神经元传播，免疫反应和 血液凝血。通过组合电化学来实现空间和暂时分辨的递送 使用纳米级电化学成像技术的化学输送系统。这种交付方式可以是 扩展到其他反应性分子，包括H2S，CO和ROS。此外，我们将建立一个精度 称为数字交付的交付工具，我们将精确控制生物分子的数量或其他非 - 通过计算其数量，包括蛋白质和纳米颗粒在内的生物实体。 以抵抗力的方式交付。最后，我们将定量绘制空间解析的摄取速率 分子传递。 最终，我们提出的研究中开发的精确交付方法将实现定量 研究与与反应性分子有关的许多基本生物学和生理问题 单细胞水平。例如，这些工具可用于揭示空间和时间异质性 神经元反应通过神经元发射器或其囊泡的精确递送。该方式也可以应用 定量调节或刺激单细胞水平的炎症反应。