In Vivo Cell Engineering by One-shot Microarrayed Electroporation for Enhanced Cancer Immunotherapy

The concept of deploying the immune system as a tool to treat cancer has been explored for a long time and becomes increasingly popular in the past decade since the approval of chimeric antigen receptor (CAR) T-cells for treating advanced blood cancer. Different cancer immunotherapies are developed with an emphasis on direct or indirect T-cell engineering around three technical routes, including immune checkpoint blockade, adoptive cell infusion, or cancer vaccination. In many cases, a first and essential process for relevant therapeutic strategy is the intracellular introduction of nucleic acid (NA)- based therapeutic materials to engineering cells of the immune system. While it is relatively easy to introduce NAs to cultured cells in laboratories, limited techniques are available for delivering NAs directly in human body, especially when the associated cytosolic delivery is required in a footprint-free manner without leaving behind any additional reagents in human body. Towards this goal, this proposal aims to address the unmet needs mentioned above by developing a revolutionary in vivo cell engineering platform. The technical innovation relies on the development of a novel hydrogel-based electronics device for in vivo, nonviral, non-liposomal, delivery of nucleic acids therapeutics to mammalian cells by microarrayed electroporation in a one-shot operation on living organisms. The multifunctional organic device (μEPO) will be made of conductive hydrogels and work as an electrical biointerface for one-shot trans-tissue local gene delivery, combining controlled macromolecules release, in situ electroporation, and in vivo gene-transfection in a trinity, which overcomes the challenges of inefficient delivery of biologic molecules (e.g., proteins, nucleic acids) to immune cells in existing immunotherapies. Based on the technical advancement, this project will further explore potential improvements of current cancer immunotherapy strategies by two different approaches, namely transdermal cancer DNA vaccination and in-body manufacturing of engineered T-cells. The first one would require subcutaneous delivery of neoantigen gene to dendritic cells for subsequent T-cell activation, offering a solution for a self-administrable and painless transdermal gene delivery in the clinical practice. The second one involves direct gene-editing in lymph node to produce immune checkpoint inhibited endogenous T-cells in living organisms in a minimally invasive manner, providing an innovative solution to augment the traditional ICB potency as a long effective way without the need of T-cell extraction and infusion. Either way, the induction tumor-specific immune response is expected to achieve prophylactic or therapeutic benefits to cancer immunotherapy in rodent animal model.