Pre-clinical development of multiplex-CRISPR-edited cell and gene therapy products

Background \u0026 Aim Data from a recent clinical trial at the University of Pennsylvania (UPenn) demonstrate the safety of NYCE T cells, which are autologous, patient-derived T cells transduced with lentivirus to express NY-ESO-1 T cell receptor and multiplex-edited with CRISPR/Cas9 at three distinct genes. This pilot study was the first in the United States (US) to administer CRISPR-edited cells to a patient. The preclinical approach and data in support of this trial was reviewed by the National Institutes of Health Recombinant DNA Advisory Committee (NIH RAC) in 2016, followed by the acceptance of an Investigational New Drug (IND) application by the US Food and Drug Administration. The aim of this abstract is to introduce the experience of an academic institution in the development of a preclinical program to support submission of an IND application for a cell therapy product with CRISPR-mediated gene editing. Non-clinical assays, data, and FDA feedback on IND submissions for CRISPR-edited gene products at the Center for Cellular Immunotherapies at UPenn will be presented. Methods, Results \u0026 Conclusion The preclinical program for the US IND application for UPenn\u0027s NYCE T cell therapy product paired conventional T cell phenotyping assays and novel genome editing evaluations. A comprehensive IND application for NYCE T cells was packaged out of inter-disciplinary collaborations to evaluate safety and potency of CRISPR-edited, antigen-directed T cells as well as fulfill NIH RAC and FDA requests. Results of these analyses will be presented. Execution of the NYCE T cell IND led to preclinical strategies toward applying CRISPR-editing to improve upon other T cell therapies, such as chimeric antigen receptor (CAR) T cells. As such, UPenn is developing an “off-the-shelf” CAR T cell product with healthy donor T cells utilizing CRISPR-mediated knock-down of genes encoding endogenous allo-reactive proteins. The parallels between the NYCE and the off-the-shelf CAR T cell non-clinical development pathways will be discussed.