Periodic Reporting for period 2 - NOVISTEM (NOn-VIral gene modified STEM cell therapy)
Berichtszeitraum: 2023-11-01 bis 2025-10-31
Despite the increasing awareness that cell and gene-therapy approaches have tremendous biomedical potential, their broad clinical application has been challenging due to prolonged and expensive production times and the emergence of severe immune- and gene-delivery dependent side effects. In this proposal, we aim to establish a stream-lined and high-throughput protocol for the generation of off-the-shelf cell therapy by combining a novel technological platform for gene delivery with a breakthrough biological concept that will permit to manufacture functional, gene-corrected blood forming stem cells and chimeric antigen receptor (CAR) T cells that can recognize and kill specific tumor cells through this CAR. To achieve this, we will optimize and apply a non-viral gene delivery method for CRISPR-mediated and site-specific gene-editing to obtain controlled CAR expression and for performing gene-correction in induced pluripotent stem cells (iPSCs). These are stem cells that have the intrinsic potential to generate any cell type of the body but that are derived from fully differentiated cells following genetic reprogramming. From these gene-modified iPSCs, we will generate CAR T cells and blood forming stem cells, respectively, by selectively targeting a signaling pathway that we established to be critical in human blood cell development and T cell development. Following functional validation of the generated cell products, we will optimize the current protocols to increase the potential for clinical implementation and establish a high-throughput platform to generate a large number of CAR expressing iPSC lines from different ages, sex and ethnicities to demonstrate the population-wide implementation potential of our approach. This will allow to generate a bank of well-characterized, CAR expressing iPSC lines that can be used to manufacture off-the-shelf cell therapy products in the absence of any rejection-related issues, thereby significantly advancing the currently implemented adaptive CAR T cell approaches by reducing the production costs and time, by selectively targeting the CAR into a well-controlled location which will prevent variability and by facilitating the production and evaluation of novel CARs for other cancer entities such as solid tumors.
We have successfully optimized photoporation as a non-viral gene delivery method to perform gene-editing in induced pluripotent stem cells iPSCs and established a detailed protocol to achieve this. We have also successfully applied a clinical-compatible protocol to generate blood forming stem cells from iPSCs and will now further establish a downstream protocol to generate T cell therapy products from them, including CAR T cells. We have successfully applied a new biological to improve the in vitro generation of T cell precursors from blood forming stem cells and also obtained important new insights into the biology of HSCs and of early and more differentiated T cell precursors that we will exploit to improve the generation and quality of our cell therapy products.
We have presented our project at several meetings, including some that activated the interest of industrial partners which resulted in novel collaborations. This has led to the successful acquisition of a novel GMP-grade compatible instrument that will allow us to test the clinical applicability of our cell therapy products and in the ability to test new biologicals to increase the success and impact of our project.