Modeling Electroporation-Mediated DNA Delivery

本更新提案寻求资金来完成在NSF拨款BES-9974185和BES-0108408下开始的电穿孔建模工作。电穿孔是一种使用强电击在细胞膜上形成孔的技术，使生物活性分子能够进入细胞内部。尽管电穿孔在药物和DNA传递方面有许多应用，但这一过程背后的理论仍然缺乏。先前由国家科学基金会资助的研究使得PI和她的团队在这一领域取得了重大进展，通过开发一个模型，该模型可以提供良好的定性，有时甚至定量，与实验一致。然而，如果没有对电穿孔的时间和空间方面进行建模，就无法达成一致的定量一致，这是本应用程序提出的研究重点。本项目的目标是：(1)对现有的孔隙形成和演化模型进行渐近简化，在不增加计算费用的情况下，得到与现有模型相当的结果。(2)建立电场驱动DNA通过大孔通量的模型，该模型可用于线性、圆形或超螺旋DNA。(3)综合单细胞和一维纤维两种典型空间扩展系统下的孔隙生成演化模型和DNA通量模型。这些模型将为细胞和组织中全面、现实的电穿孔模型的可行性提供概念证明，并为体内电穿孔的临床过程建模奠定基础。由Wanda博士组成的跨学科研究团队将进行研究。约翰·纽博士（加州大学伯克利分校数学系），研究生研究助理，以及本科生。该模型将由Genetronics公司的研发科学家每年进行评估。Genetronics公司是一家专门从事电穿孔医疗应用技术和硬件开发的公司。

0401757KrassowskaThis renewal proposal seeks funding to complete the work on modeling electroporation that wasstarted under the NSF grants BES-9974185 and BES-0108408. Electroporation is a technique in which strong electrical shocks are used to create pores in the cell membrane that enable biologically active molecules to enter the cell interior. Despite numerous applications of electroporation to drug and DNA delivery, the theory underlying this process is still lacking. Previous research funded by the NSF allowed the PI and her team to make significant progress in this area, by development of a model that gives good qualitative, and sometimes even quantitative, agreement with experiments. However, consistent quantitative agreement cannot be reached without modeling both temporal and spatial aspects of electroporation, which is the focus of the research proposed in this application. The goals of this project are:(1) Develop an asymptotic simplification of the existing model of pore creation and evolution, which would give results equivalent to the existing model without its computational expense.(2) Develop a model of electric field-driven DNA flux through macropores that could be used forlinear, circular, or supercoiled DNA. (3) Synthesize the pore creation and evolution model and the DNA flux model in the context of two prototypical spatially extended systems, a single cell and a one-dimensional fiber. These models will provide the proof of concept that a comprehensive and realistic model of electroporation in cells and tissues is feasible, and they will lay the foundation for modeling clinical procedures involving in-vivo electroporation.An interdisciplinary research team consisting of Dr. Wanda will perform the researchKrassowska (PI, Biomedical Eng., Duke University), Dr. John Neu (Mathematics, UC Berkeley),a graduate research assistant, and undergraduate students. The model will be evaluated yearlyby the R&D scientists from Genetronics, Inc., a company that specializes in developing technology and hardware for medical applications of electroporation.