PROJECT SUMMARY – CORE C Core C: cGMP Manufacturing Core (cGMP Core) serves as a full service "turn key" vector and cell manufacturing for the PPG by providing cGMP lentiviral vectors and transduced CD34+ cells (Drug Product) for clinical trials in all three projects. The function of the cGMP Core is the development, scale-up, and cGMP manufacturing of vector and the assays required for release and characterization of the lentiviral vectors and the development, scale-up, cGMP manufacturing and the assays required for release and characterization of transduced CD34+ cells from bone marrow or apheresis collections. The cGMP Core must work directly with the each of the Project PIs and the Vector Core as the complexity of these products requires a highly orchestrated and well-thought out project timeline that takes into account all of the aspects defined in a Target Product Profile The cGMP Core is tightly integrated with the Vector Development Core and the PI's laboratories and meets with both groups every two weeks to discuss development and production activities. This allows the other investigators to test vectors and cell products to insure biological potency and functionality. Once a cGMP process is ready for transition into clinical trial, the investigators and/or Vector Core transfer the vector and product-specific bioanalytical methods to the cGMP Core for process and analytical methods development, process scale-up, and cGMP manufacturing. Examples of this integration during that past funding period include the first clinical use of the GPRG lentiviral packaging cell line for use in the LVXSCID trials and the recent transfer of the second generation stable producing cell line for WAS. A similar process is anticipated for development and production of a lentiviral vector for sickle cell disease and will likely require the use of transient transfection for cGMP production. Other achievements in the past funding period have been the development and clinical implementation of bone marrow processing and transduction batch records for the infant XSCID trial, which has now been successfully used in four infant cases. The cGMP Core has over 10 years of experience and Dr. Meagher (leader of the cGMP Core) has 30 years of experience in the field and will oversee the development and optimization of improved and scalable technology for production of lentivirus (LV), genetically modified peripheral blood and bone marrow CD34+ cells, and eventually genome editing technologies including AAV vectors and recombinant genome editing enzymes as requested by the PIs. The cGMP Core will also work with the new Experimental Cellular Gene Therapy (ECGT) Core at St. Jude on the development and scale-up of the transduction of CD34+ cells. All cGMP products will be manufactured by the cGMP Core.

PROJECT NARRATIVE – CORE C The cGMP Core is essential for development and cGMP manufacturing of vectors, cell processing of transduced cells, and later, gene editing of CD34+ cells in support of clinical gene therapy protocols.