Multiscale Modelling of Microfluidic Cell Mechanoporation for Intracellular Gene Delivery

Intracellular gene delivery has ground-breaking applications in mRNA vaccines, preparation of CAR-T cells for cancer treatment, and genetic engineering of plants. Microfluidic-based delivery, which uses fluid flow to disrupt cell membranes, leading to transient formation of membrane pores and internalisation of genetic materials, is emerging as a simple, universal, and transformative approach compared with classical methods that utilise electroporation or viral vectors. However, development of microfluidic-based approaches has been conducted largely through trial-and-error experiments, due to the lack of a high-fidelity predictive simulation tool. This interdisciplinary fellowship will integrate the fellow’s strong expertise in modelling fluid-structure interaction and nanoscale membrane damage, with the host’s experimental expertise in cell mechanics and microfluidic intracellular delivery. It will develop an accurate predictive simulation tool and fundamental understanding of microfluidic flow-induced cell poration, and the associated internalisation of exogenous genetic materials. Specific scientific aims are: 1, to develop a novel three-dimensional multiscale model, to enable quantitative predictions of cell deformation, membrane mechanoporation and intracellular delivery of genetic materials in microfluidics; 2, to conduct systematic experiments, measuring cell membrane pores and gene concentration inside cells, and use the data to validate the model; 3, to combine simulations and experiments to uncover the effects of flow condition, cell mechanical property and molecular weight of genetic materials, on cell membrane poration, cell viability and gene-delivery efficiency. Through carefully planned knowledge transfer, dissemination and trainings, the fellowship will significantly enhance the fellow’s career perspective for establishing independence, and contribute to the EU Research and Innovation Strategy in improving and protecting people's health.