Research Project
9622672
PROJECT SUMMARY Dendritic cells (DC) are antigen-presenting cells that identify and capture foreign bodies and play an essential role in activating both the adaptive and humoral immune response. DCs are an attractive vehicle for therapeutic manipulation, particularly in cancer, and are used in antigen-pulsed autologous DC therapies and DC-stimulated autologous T cell therapies. However, direct isolation of patient-specific DCs is challenging since they are present in very low concentrations (<1%) in human blood and their ex-vivo generation using monocytes or stem cell precursors using the industry standard manual culture is time-consuming, labor- intensive and subject to user-variability. The manual technique remains the standard even in clinical manufacturing (e.g. for the first DC vaccine Provenge, which currently treats around 4,000 patients per year in the US) because no effective system exists for automated dendritic cell generation in a single-use, patient- specific format. Yet, a significant market exists for such automated systems. There are >120 clinical trials currently ongoing clinical trials in new, precisely targeted dendritic cell vaccines and T cell therapies produced using patient-specific and tumor-specific neoantigen-stimulated DCs. To meet this significant clinical and market need, Flaskworks is developing a suite of proprietary automated systems, including MicroDEN, a perfusion-based automated culture system for patient-specific DC generation. In its current form, the MicroDEN cell culture cartridges consist of untreated polystyrene which has limited capacity to adhere monocytes. In this proposal, we describe an approach to engineer the surface of the MicroDEN cartridges to significantly increase monocyte adhesion and thereby DC yield, a crucial attribute for clinical manufacturing. MicroDEN Plus cartridges will be designed by mimicking key aspects of the in vivo environment, including presentation of ECM signals to enhance monocyte adhesion and drive their conversion to DCs. Given that no automated, single use system of this kind is currently available, we expect the MicroDEN Plus system to have significant impact in the manufacturing of DCs for clinical use in DC vaccines and DC-stimulated T cell therapies.
