ERI: Revealing the Reconfiguration Dynamics of Lipid Bilayer and Its Hydration Structures with Nanoscale Resolution during Electroporation

生命科学中的一个基本问题是了解和控制细胞膜对外界刺激的反应。这对于确定药物输送的有效性是很重要的。例如，电场可以在细胞膜上产生瞬时的气孔，这是一种被称为电穿孔的过程，用于将货物运送到细胞内，但要在不造成不可逆转的损害的情况下实现有效的运送仍然是一个挑战。该奖项将使用亚纳米分辨率成像来捕捉细胞膜在施加外部电场时如何动态响应。对细胞内给药的了解不仅有助于生物医学领域，还将有助于生物分子的制造、基因编辑等细胞内研究。这种成像能力还将有助于理解燃料电池、电池和腐蚀/腐蚀过程等应用中的固液界面系统。与研究活动相配合，生物界面和成像技术的知识将通过开发和使用移动和网页应用程序进行工作流程可视化，向K-12学生和老年住院医生进行生物界面演示，以及为本科生和研究生提供培训，向公众传播。该奖项将使用亚纳米和高时间分辨率的原子力显微镜来提供分子水平的成像，重点在于揭示脂质双层及其水化结构在外部电场下的动态响应。由于缺乏合适的成像工具，对药物传递过程中涉及的纳米级相互作用的了解仍然有限。这项工作的关键假设是，脂质双层的水化结构决定了脂质双层与药物分子的纳米级相互作用以及在外界刺激下脂质双层的生物物理行为。将启动三个研究方向：i)成像电场下脂双层的动态变化；ii)捕捉参与脂双层重组的水化结构的结构变化；iii)建立不同类型脂类和溶液化学的结构-性质关系。建议的研究活动旨在解决和理解脂质双层电穿孔过程中的孔形成和封闭过程，并揭示水化结构在这一过程中的作用。拟议的研究和教育活动将弥合生物界面分子现象分子水平成像方面的长期知识差距，并为更好地设计用于生物医学目的的非病毒转基因方法提供见解。该奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

A fundamental question in life science is to understand and control how cell membranes respond to external stimuli. This is important in determining the efficacy of drug delivery. For example, electric fields can create transient pores on a cell membrane, a process called electroporation, which is used to deliver cargo into cells, but it remains a challenge to achieve efficient delivery without causing irreversible damage. This award will use sub-nanometer resolution imaging to capture how cell membranes dynamically respond upon applying an external electric field. The understanding of intracellular delivery gained will not only benefit the biomedical field, but also biomolecule manufacture, gene editing and other intracellular investigations. This imaging capability will also benefit the understanding of solid-liquid interface systems in applications including fuel cells, batteries, and corrosion/erosion processes. In synergy with the research activities, the knowledge of bio-interfaces and imaging techniques will be disseminated to the general public, by developing and using mobile and webpage apps for workflow visualization, doing demos of bio-interfaces to K-12 students and senior residents, and providing training to both undergraduate and graduate students. This award will use an atomic force microscope with sub-nanometer and high temporal resolution to offer the molecular-level imaging, with a focus on revealing the dynamic responses of lipid bilayer and its hydration structure under an external electric field. The understanding of nanoscale interactions involved in the drug delivery process are still limited due to lack of appropriate imaging tools. The key hypothesis in this work is that the hydration structure of lipid bilayer determines the nanoscale interactions of lipid bilayer with drug molecules and the biophysical behavior of lipid bilayer under the external stimuli. Three research thrusts will be initiated: i) imaging the dynamic changes of lipid bilayer under an electric field; ii) capturing the structural changes of hydration structures involved in lipid bilayer reconfiguration; and iii) establishing the structure-property relationship for different types of lipid and solution chemistry. The proposed research activities aim to resolve and understand the pore formation and sealing processes during electroporation of lipid bilayers and reveal the role of hydration structure in this process. The proposed research and education activities will bridge the long-standing knowledge gap in the molecular-level imaging of molecular phenomena at bio-interfaces and offer insights into better engineering of non-viral transfection methods for biomedical purposes.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.