Application of pulsed electric field (PEF) can reversibly increase the permeability of the cell membrane allowing the access of otherwise impermeable DNA molecules to the inside of the cell. Introduction of foreign DNA molecules encoding immuno-modulatory proteins, antibodies and antigens into cells using PEF, known as Gene Electro-Transfer (GET), is increasingly used for the modulation of the immune system or immunotherapy. While GET based immunotherapy presents itself as a potent application for treatment of cancer and vaccination against infectious diseases, it is suffering from low levels of transgene expressions in vivo. This low efficiency can largely be attributed to our lack of fundamental understanding of the mechanisms by which DNA molecules overcome the barriers of the extra-cellular matrix and the cell membrane in the presence of an electric field. In this action, I aim to provide this required fundamental understanding using principles of polymer physics, soft matter and statistical mechanics. Experiments based on these principles will be conducted in vitro and in vivo to generate results that can be directly compared to theories and models of DNA transport through the extra-cellular matrix and the cell membrane. Understanding the mechanisms within the frameworks of polymer physics will radically improve the efficiency of GET immunotherapy, because it will provide a mechanistic ground for developing optimum protocols within complex tissue environments that can, at the same time, be readily transferred across tissue types and species.
Cardiovascular diseases are the No. 1 healthcare challenge in the world, among which ischemic heart disease and atrial fibrillation are the most prevalent. Better treatment strategies are greatly needed to reduce the medical, economic, and social burden of these conditions. Electroporation (application of intense pulsed electric field) is showing tremendous potential for treatment of atrial fibrillation, enabling a safer and shorter treatment procedure compared with existing thermal ablation approaches. Moreover, recent pioneering studies provide evidence that electroporation can also be used as a nonviral vector for intracellular delivery of therapeutic nucleic acids that promote cardiac regeneration, potentially offering a way to cure the so-far incurable ischemic heart disease. For treatment of atrial fibrillation, electroporation must be irreversible, resulting in the death of cardiac muscle cells, to locally destroy (ablate) the arrhythmogenic cardiac tissue. Conversely, for treatment of ischemic heart disease electroporation must be reversible, meaning that the pulsed electric field transiently enhances cellular uptake of nucleic acids while the cells are able to survive and express the delivered transgene(s). Due to a lack of fundamental understanding of cardiac electroporation, there are currently no reliable methods able to ensure electroporation (ir)reversibility and the desired treatment outcome. This project is designed to decipher the biophysical mechanisms of cardiac electroporation at the molecular, cellular and tissue level as to develop methodologies that will enable optimal implementation of both irreversible and reversible electroporation. By combining bottom-up experiments in primary cardiac cells and tissue slices with computational modeling and advanced data analysis I will create the foundations needed to streamline further (pre)clinical research and realize the potential of electroporation to advance cardiac treatments.
Charged colloidal suspensions consist of charged micron-sized particles dispersed in a solvent with nanometer-sized ions, and they find ample applications in material science, food industry and drug development. The particle charge, together with a diffuse ion cloud that screens this particle charge, is called the electric double layer and it is pivotal in understanding the phase behaviour and interactions in these suspensions. However, no attention has been paid to the topological properties of the electric double layer. While drawing heavily on recent advances in liquid crystalline systems with topologically non-trivial orientational ordering, we propose to explore the topology of electric double layers, which could ultimately lead to enhanced stability of charged colloidal crystals. Specifically, we will focus on particles with topologically non-trivial shapes and explore the coupling between the topological invariants of the particle (such as genus) with the emergent electric double layer, and how this affects interparticle interactions and the phase behaviour. Finally, the goal of this proposal is to obtain a precise control over charged colloidal suspensions that are protected by topological, rather than only energetic binding, opening a fundamental and applied route to a new class of topological soft matter.