EAGER: Biomanufacturing: Physiologically-inspired Large Scale Manufacturing and Potency-Biomarker identification for Chimeric Antigen Receptor (CAR)-T cells

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1547638 Roy, KrishnenduA novel, biologically-inspired strategy is proposed to enhance the expansion of therapeutic T cells by mimicking the environment and conditions of lymph nodes, where these cells normally expand in the body. This idea of T cell expansion inside synthetic lymph-node-like niches, where cells communicate with each other and with 'self' is unique, but firmly rooted on physiological principles. Our approach could vastly enhance the expansion potential of T cells making larger numbers of them available for therapy and also reduce overall manufacturing cost, making the treatment more affordable and widely available. Further, predictively measuring the efficacy and potency of these manufactured T cells, before infusing them in patients, is of critical importance. A comprehensive characterization of manufactured T cells will be carried out to develop robust correlative markers that can predict cell potency and can eventually be used as a routine analysis tool to determine the quality of these cells before infusion.This EAGER project will investigate human CAR-T cell expansion within synthetic 3D microenvironments that mimic the T cell niches within lymph nodes (LN). Anti-CD3/anti-CD28-functionalized microcarriers will be used and human CAR-T cells will be cultured in LN-mimicking 3D niches where T cells remain at high density with close cell-cell contact, and allow efficient paracrine/autocrine signaling. These parameters, absent from the current T-cell manufacturing concepts, are critical since T cells secrete large amounts of interleukins (ILs) locally to promote rapid, large scale expansion. Thus, this approach could also reduce culture media and IL requirements, thereby significantly reducing cost. The effect of low O2 tension will be evaluated together with various cell-seeding densities and anti-CD3/anti-CD28 ligand densities on expansion efficacy and T cell quality. Further, the LN-like niche will be combined with stirred tank or perfusion bioreactors, to determine whether dynamic culture and flow perfusion improve expansion efficacy, product quality, scalability and cost effectiveness.