Cell Processing Section, Department of Transfusion Medicine, Clinical Center, National Institutes of Health, Bethesda, Maryland 20892, USA.
Mol Ther. 2013 Feb;21(2):476-84. doi: 10.1038/mt.2012.89. Epub 2012 Nov 13.
Cell-based immunotherapies are among the most promising approaches for developing effective and targeted immune response. However, their clinical usefulness and the evaluation of their efficacy rely heavily on complex quality control assessment. Therefore, rapid systematic methods are urgently needed for the in-depth characterization of relevant factors affecting newly developed cell product consistency and the identification of reliable markers for quality control. Using dendritic cells (DCs) as a model, we present a strategy to comprehensively characterize manufactured cellular products in order to define factors affecting their variability, quality and function. After generating clinical grade human monocyte-derived mature DCs (mDCs), we tested by gene expression profiling the degrees of product consistency related to the manufacturing process and variability due to intra- and interdonor factors, and how each factor affects single gene variation. Then, by calculating for each gene an index of variation we selected candidate markers for identity testing, and defined a set of genes that may be useful comparability and potency markers. Subsequently, we confirmed the observed gene index of variation in a larger clinical data set. In conclusion, using high-throughput technology we developed a method for the characterization of cellular therapies and the discovery of novel candidate quality assurance markers.
基于细胞的免疫疗法是开发有效和靶向免疫反应的最有前途的方法之一。然而,它们的临床用途和疗效评估严重依赖于复杂的质量控制评估。因此,迫切需要快速系统的方法来深入表征影响新开发的细胞产品一致性的相关因素,并确定用于质量控制的可靠标志物。本研究以树突状细胞(DC)为模型,提出了一种全面表征制造细胞产品的策略,以确定影响其变异性、质量和功能的因素。在生成临床级别的人单核细胞来源的成熟树突状细胞(mDC)后,我们通过基因表达谱分析测试了与制造过程相关的产品一致性程度以及由于个体内和个体间因素引起的可变性,以及每个因素如何影响单个基因的变异。然后,通过计算每个基因的变异指数,我们选择了用于身份检测的候选标志物,并定义了一组可能对可比性和效力标志物有用的基因。随后,我们在更大的临床数据集上验证了观察到的基因变异指数。总之,我们使用高通量技术开发了一种用于细胞治疗表征和发现新的候选质量保证标志物的方法。