Modeling Electroporation-Mediated DNA Delivery

0401757 Krassowska这一更新提案寻求资金，以完成在NSF赠款BES-9974185和BES-0108408下开始的电穿孔建模工作。电穿孔是一种技术，其中使用强电击在细胞膜中产生孔，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递送中有许多应用，但仍然缺乏这一过程的理论基础。由NSF资助的先前研究使PI和她的团队在这一领域取得了重大进展，通过开发一种模型，该模型提供了良好的定性，有时甚至是定量，与实验一致。然而，如果不对电穿孔的时间和空间方面进行建模，则无法达到一致的定量协议，这是本申请中提出的研究的重点。本计画的目标是：（1）发展一个渐进简化的孔隙产生与演化的现有模型，其结果将等同于现有的模型，而没有其计算费用。(2)开发一个电场驱动的DNA通过大孔的模型，可用于线性，环形或超螺旋DNA。(3)在两个典型的空间扩展系统（单细胞和一维纤维）的背景下，综合孔的产生和演化模型以及DNA通量模型。这些模型将为细胞和组织中电穿孔的全面和现实模型的可行性提供概念证明，并且它们将为涉及体内电穿孔的临床程序建模奠定基础。由Wanda博士组成的跨学科研究团队将进行研究Krassowska（PI，Biomedical Eng.，杜克大学），博士约翰Neu（数学，加州大学伯克利分校），研究生研究助理，和本科生。该模型将每年由Genetronics公司的研发科学家进行评估，一家专门开发电穿孔医疗应用技术和硬件的公司。

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.