Establishing the POROS Intracellular Delivery System as a Platform for Cellular Therapeutics

Abstract CAR-T therapy is a rapidly emerging therapy for the treatment of cancer, with two companies receiving FDA approval for their first CAR-T therapies in 2017. These therapies use immune cells called T cells, either from the patient themselves, or a healthy donor, and genetically engineer them to kill the tumor cells. The genetic engineering involves direct delivery of nucleic acids to these cells, which is challenging as current delivery methods either lack efficiency, are very damaging to cells, or both. This engineering is performed on a case- by-case basis and the methods are costly and time-consuming. Next generation CAR-T therapies will include more complex genetic engineering, with two or more editing events required. This is currently being done in a sequential fashion, which increases the time-to-patient of the final product. In order to make CAR-T therapies available to more patients in a cost-effective, safe and timely manner, new approaches need to be developed. Current research is addressing this need by developing new technologies for cell treatment that allow faster recovery and expansion time of the treated cells, allow delivery of multiple molecules in one shot and that can be integrated into a closed, manufacturing-ready system. Another focus of research is engineering healthy donor T cells to be used in CAR-T therapy, which would allow for an off-the-shelf cellular therapeutic. For this type of product to be realized, high capacity treatment methods need to be developed capable of treating rt 1010 cells while also maintaining high cell viability and delivery efficiency. OpenCell Technologies has developed a proprietary technology, POROS, to deliver macromolecules such as DNA, RNA and protein to a wide variety of cell types. POROS uses an acoustic drive to push cells through an array of nozzles one cell at a time, thus creating shear force that porates the cells. In this SBIR program, OpenCell is expanding the capabilities of its POROS platform by developing higher capacity POROS devices. Approach: In this Phase II project, OpenCell will develop a high capacity version of POROS, capable of treating a larger number of cells to enable CAR-T development. We will develop a prototype POROS device, capable of treating 1-5 × 108 cells in under 10 minutes, and demonstrate gene editing in T cells as a proof-of- concept experiment. The POROS device development will pave the way for development of a large, all-in-one, GMP-compliant POROS-giga device, for manufacturing of off-the-shelf cellular therapeutics. Additionally, the POROS unit developed in this Phase II project will also be a salable unit of its own with anticipated applications in research and development, personalized medicine and small batch production of cells for autologous cell therapies and for clinical trials.