Point-Of-Care CAR T-Cell Production (ARI-0001) Using a Closed Semi-automatic Bioreactor: Experience From an Academic Phase I Clinical Trial
Point-Of-Care CAR T-Cell Production (ARI-0001) Using a Closed Semi-automatic Bioreactor: Experience From an Academic Phase I Clinical Trial
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DOI：
10.3389/fimmu.2020.00482
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发表时间：
2020-03-20
影响因子：
7.3
通讯作者：
Juan, Manel
中科院分区：
文献类型：
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作者：
Castella, Maria;Caballero-Banos, Miguel;Juan, Manel
开发半自动化设备，可以减少动手时间并标准化临床级CAR T细胞的生产，例如Miltenyi的CliniMACS Prodigy，是促进CAR T细胞疗法开发的关键，特别是在学术机构。然而，使用该系统从重度预治疗患者中生产CAR T细胞产品的可行性尚未得到证实。在这里，我们报告并表征了在CD 19 + B细胞恶性肿瘤的I期临床试验（NCT 03144583）的背景下28种CAR T细胞产物的产生。该系统包括CD 4-CD 8细胞选择、慢病毒转导和使用IL-7/IL-15的T细胞扩增。生产的28种CAR T细胞产品中有27种符合完整的质量标准列表，被视为有效产品。离体细胞扩增平均持续8.5天，平均转导率为30.6 +/-13.44%。获得的所有产物均呈现出针对CD 19+细胞的细胞毒性活性，并且精通促炎细胞因子的分泌。与健康供体细胞相比，患者细胞的扩增动力学较慢。然而，产品效价相当。CAR T细胞亚群表型在患者中高度可变，主要由初始产物决定。T-CM和T-EM是获得的主要T细胞表型。平均而言，获得的38.7%的CAR T细胞呈现T-N或T-CM表型，这是能够在患者中建立持久T细胞记忆的亚群。一项深入分析，以确定有助于最佳T细胞表型的各个因素，揭示了离体细胞扩增导致T-N，T-SCM和T-EFF细胞数量减少，而T-CM细胞增加，这两者都是由于细胞扩增和CAR表达。总体而言，我们的研究结果首次表明，使用CliniMACS Prodigy系统为重度预治疗患者生产临床级CAR T细胞是可行的，并且获得的产品符合该领域的现行质量标准。减少的离体扩增可以产生具有增加的体内持久性的CAR T细胞产物。
Development of semi-automated devices that can reduce the hands-on time and standardize the production of clinical-grade CAR T-cells, such as CliniMACS Prodigy from Miltenyi, is key to facilitate the development of CAR T-cell therapies, especially in academic institutions. However, the feasibility of manufacturing CAR T-cell products from heavily pre-treated patients with this system has not been demonstrated yet. Here we report and characterize the production of 28 CAR T-cell products in the context of a phase I clinical trial for CD19+ B-cell malignancies (NCT03144583). The system includes CD4-CD8 cell selection, lentiviral transduction and T-cell expansion using IL-7/IL-15. Twenty-seven out of 28 CAR T-cell products manufactured met the full list of specifications and were considered valid products. Ex vivo cell expansion lasted an average of 8.5 days and had a mean transduction rate of 30.6 +/- 13.44%. All products obtained presented cytotoxic activity against CD19+ cells and were proficient in the secretion of pro-inflammatory cytokines. Expansion kinetics was slower in patient's cells compared to healthy donor's cells. However, product potency was comparable. CAR T-cell subset phenotype was highly variable among patients and largely determined by the initial product. T-CM and T-EM were the predominant T-cell phenotypes obtained. 38.7% of CAR T-cells obtained presented a T-N or T-CM phenotype, in average, which are the subsets capable of establishing a long-lasting T-cell memory in patients. An in-depth analysis to identify individual factors contributing to the optimal T-cell phenotype revealed that ex vivo cell expansion leads to reduced numbers of T-N, T-SCM, and T-EFF cells, while T-CM cells increase, both due to cell expansion and CAR-expression. Overall, our results show for the first time that clinical-grade production of CAR T-cells for heavily pre-treated patients using CliniMACS Prodigy system is feasible, and that the obtained products meet the current quality standards of the field. Reduced ex vivo expansion may yield CAR T-cell products with increased persistence in vivo.