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

EAGER：生物制造：嵌合抗原受体 (CAR)-T 细胞的生理学启发大规模制造和效力生物标志物鉴定

• 批准号: 1547638
• 负责人: Krishnendu Roy
• 金额: $ 30万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2018-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1547638&HistoricalAwards=false
• 关键词: EAGER; Biomanufacturing; Physiologically; inspired; Large

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.

1547638 Roy，Krishnovich提出了一种新的生物启发策略，通过模拟淋巴结的环境和条件来增强治疗性T细胞的扩增，这些细胞通常在体内扩增。这种T细胞在人造淋巴结样壁龛内扩增的想法是独特的，但牢固地植根于生理学原理。我们的方法可以极大地增强T细胞的扩增潜力，使更多的T细胞可用于治疗，并降低整体制造成本，使治疗更实惠，更广泛地使用。此外，在将这些制造的T细胞注入患者之前，预测性地测量它们的功效和效力是至关重要的。将对制造的T细胞进行全面表征，以开发出强大的相关标记物，这些标记物可以预测细胞的效力，并最终可以用作常规分析工具，在输注前确定这些细胞的质量。EAGER项目将研究人类CAR-T细胞在模拟淋巴结（LN）内T细胞小生境的合成3D微环境中的扩增。将使用抗CD 3/抗CD 28功能化的微载体，并将在LN模拟3D壁龛中培养人CAR-T细胞，其中T细胞保持高密度，细胞-细胞接触紧密，并允许有效的旁分泌/自分泌信号传导。这些参数在当前的T细胞制造概念中是不存在的，但这些参数是关键的，因为T细胞局部分泌大量的白细胞介素（IL）以促进快速、大规模扩增。因此，这种方法还可以减少培养基和IL需求，从而显著降低成本。将评价低O2张力以及各种细胞接种密度和抗CD 3/抗CD 28配体密度对扩增效力和T细胞质量的影响。此外，LN样生态位将与搅拌罐或灌注生物反应器组合，以确定动态培养和流动灌注是否改善扩增功效、产品质量、可扩展性和成本效益。