CORE C: Summary Abstract Core C is comprised of two groups: the Human Genome Editing Laboratory (HGEL) led by Dr. Friederike Herbst and the Translational and Correlative Studies Laboratory (TCSL) led by Dr. Joseph A. Fraietta. Core C works closely with Cores A and B to actively support the proposed P01 grant renewal with the following three Aims: In Specific Aim 1: Process Optimization, Development and Product Characterization, Core C will provide knowhow and consulting support to Core B to manufacture CRISPR-edited CAR T cells in a shortened manufacturing process (Project 1: CD5 knockout anti-CD19 CAR T cells). Core C will conduct scale-up studies to optimize base editing of T cells and matching HSPC cell populations for further lockdown of manufacturing processes to tech transfer to Core B (Project 2). Core C will develop a pipeline for subsequent characterization of the gene edited cell products (T cells and HSPC) including assay developmental procedures important for product safety and release testing, in line with FDA regulations for future IND applications. In Specific Aim 2: Support of Clinical Trials Proposed by the 3 Projects, Core C will support all three clinical trials within this grant proposal. Project 1: CD5 knockout anti-CD19 CAR T cells in patients with relapsed or refractory lymphoid malignancies. Project 2: CD45-edited CART45 in tandem with CD45-edited matching HSPCs in patients with acute myeloid leukemia; and Project 3: combination of MIL- and PBL-derived anti-BCMA CAR T cells in patients with relapsed/refractory multiple myeloma. In addition to supporting the manufacturing Core B with product safety analysis and product release assays testing for on-/off target editing, transduction efficiency, replication- competent lentivirus and mycoplasma, Core C will perform a range of sample processing, biobanking and correlative assays and data collection that will be tailored to the individual needs of each of the clinical trials. In Specific Aim 3: Analytical Platform Development, Core C will strengthen three assay platforms identified as being key correlative assay technologies needed to support the projects and clinical trials in this grant proposal. Overall, all platforms - once established - will allow the identification as well as the molecular and phenotypic characterization of rare subpopulations of therapeutic cells. Moreover, the generated data package will be evaluated and utilized to predict and improve patient outcome. 1. Single cell transcriptome analysis to expand correlative analysis by extending established protocols and existing data to optimize single-cell methods on samples drawn from internally biobanked accessions. 2. Digital PCR using a new QIAcuity One plate-based system to perform sensitive standard-independent quantification of rare sequences and support the trials with assessment of gene editing and gene marking. 3. Flow cytometry Developing new 28+ color panels on a spectral cytometer to be used for retros...