Nanoscale probes for sensing molecular functions in live cells

用于感测活细胞中分子功能的纳米级探针

• 批准号: 10201347
• 负责人: Bianxiao Cui
• 金额: $ 81.45万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10201347
• 关键词: Action Potentials; Affect; Biochemical; Biocompatible Materials; Biological; Biotechnology; Cardiac Myocytes; Cell physiology; Cells; Classification; Development; Electrodes; Electrophysiology (science); Engineering; Environment; Extracellular Matrix; Future; Generations; Goals; In Vitro; Intervention; Knowledge; Mechanics; Membrane; Molecular; Monitor; Nanotechnology; Nanotopography; Process; Proteins; Role; Rupture; Signal Transduction; Surface; base; biological systems; design; heart cell; mechanical force; nanoelectrodes; nanoscale; parent grant; patch clamp; sensor; stem cells; tool; transmission process

Project Summary / Abstract: This MIRA proposal merges two distinct projects supported by R01GM128142, “The role of membrane curvature in surface nanotopography-induced cell functions”, and R01GM125737, “Developing nanoscale electrophysiology sensors for robust intracellular recording”. While the two projects focus on different biological questions, the unifying theme is to develop nanoscale probes to elucidate the cellular machinery in the intricate environment of living cells. In this proposal, we discuss topics along the lines of the parent grants, focusing on the significance of the biological problems, our recent and evolving results, and directions for the future. For the first project, the long-term goal is to understand how membrane curvature regulates biochemical signals that are transmitted through the cell-matrix interface. At the cell-matrix interface, where the cells make physical contact with extracellular matrices, the membrane may be locally deformed by matrix topography or mechanical forces. As it remains a challenge to manipulate nanoscale membrane curvature in live cells, our current understanding of how local membrane curvature affects signal transmission is limited. We propose to use nanotechnology-based precision engineering to control interface membrane curvature in live cells. We seek to understand how cellular processes are affected by membrane curvature and the underlying molecular mechanisms. The knowledge gained will help us understanding how cells interact with extracellular matrix and also help us designing biomaterials for better integration with cells. For the second project, we are developing vertical nanoelectrodes into a robust and easy-to-use electrophysiology tool that can reliably achieve parallel intracellular recording of cardiomyocytes with minimal perturbation. Simultaneous nanoelectrode and patch clamp recordings on same cells confirmed that nanoelectrodes accurately record action potential waveforms for classification and characterization of stem-cell-derived cardiomyocytes. These nanoelectrodes will enable us to understand how in vitro interventions accelerate the maturation of stem-cell-derived cardiomyocyte. Furthermore, nanoelectrodes provide an ideal tool for monitoring the generation and resealing of membrane pores on cardiomyocytes that are prone to membrane rupture due to their large size and strong mechanical contraction. We will use nanoelectrode to investigate how proteins participate in the membrane resealing process. We hope to achieve a broad impact by combining the development of new tools with applications to specific biological systems.

项目概要/摘要： 该MIRA提案合并了R 01 GM 128142支持的两个不同项目，“膜的作用 曲率在表面纳米地形诱导细胞功能”，和R 01 GM 125737，“开发纳米 用于稳健的细胞内记录的电生理学传感器”。虽然这两个项目侧重于不同的生物 问题，统一的主题是开发纳米探针，以阐明复杂的细胞机制， 活细胞的环境。在本建议书中，我们沿着母公司赠款金的思路讨论主题，重点是 生物学问题的重要性，我们最近和不断发展的结果，以及未来的方向。为 第一个项目，长期目标是了解膜曲率如何调节生化信号 通过细胞-基质界面传输。在细胞-基质界面上，细胞在那里形成物理 与细胞外基质接触时，膜可能会因基质形貌而局部变形， 机械力由于在活细胞中操纵纳米级膜曲率仍然是一个挑战， 目前对局部膜曲率如何影响信号传输的理解是有限的。我们建议 使用基于纳米技术的精密工程来控制活细胞中的界面膜曲率。我们 试图了解细胞过程是如何受到膜曲率和潜在分子的影响， 机制等所获得的知识将有助于我们了解细胞如何与细胞外基质相互作用， 也帮助我们设计生物材料，使其更好地与细胞结合。对于第二个项目，我们正在开发 垂直纳米电极转换成一个强大的和易于使用的电生理工具，可以可靠地实现平行 心肌细胞的细胞内记录具有最小的扰动。纳米电极和贴片同步 在相同细胞上的钳记录证实纳米电极准确地记录动作电位波形 用于干细胞衍生的心肌细胞的分类和表征。这些纳米电极将使 让我们了解体外干预如何加速干细胞衍生的心肌细胞的成熟。 此外，纳米电极为监测膜的生成和重新密封提供了理想的工具 心肌细胞上的孔由于它们的大尺寸和强机械性能而易于膜破裂， 收缩。我们将使用纳米电极来研究蛋白质如何参与膜的重新密封 过程我们希望通过将新工具的开发与应用程序相结合， 特定的生物系统。