Sarén Tina, Ramachandran Mohanraj, Gammelgård Gustav, Lövgren Tanja, Mirabello Claudio, Björklund Åsa K, Wikström Kristina, Hashemi Jamileh, Freyhult Eva, Ahlström Håkan, Amini Rose-Marie, Hagberg Hans, Loskog Angelica, Enblad Gunilla, Essand Magnus
Department of Immunology, Genetics and Pathology, Uppsala University, Science for Life Laboratory, Uppsala, Sweden.
IFM Bioinformatics, National Bioinformatics Infrastructure Sweden, Science for Life Laboratory, Linköping University, Linköping, Sweden.
Clin Cancer Res. 2023 Oct 13;29(20):4139-4152. doi: 10.1158/1078-0432.CCR-23-0178.
Although CD19 chimeric antigen receptor T cells (CAR-T) therapy has shown remarkable success in B-cell malignancies, a substantial fraction of patients do not obtain a long-term clinical response. This could be influenced by the quality of the individual CAR-T infusion product. To shed some light on this, clinical outcome was correlated to characteristics of CAR-T infusion products.
In this phase II study, patients with B-cell lymphoma (n = 23) or leukemia (n = 1) received one or two infusions of third-generation CD19-directed CAR-Ts (2 × 108/m2). The clinical trial was registered at clinicaltrials.gov: NCT03068416. We investigated the transcriptional profile of individual CD19 CAR-T infusion products using targeted single-cell RNA sequencing and multicolor flow cytometry.
Two CAR-T infusions were not better than one in the settings used in this study. As for the CAR-T infusion products, we found that effector-like CD8+CAR-Ts with a high polyfunctionality, high cytotoxic and cytokine production profile, and low dysfunctional signature were associated with clinical response. An extended ex vivo expansion time during CAR-T manufacturing negatively influenced the proportion of effector CD8+CAR-Ts in the infusion product.
We identified cell-intrinsic characteristics of effector CD8+CAR-Ts correlating with response that could be used as an indicator for clinical outcome. The results in the study also serve as a guide to CAR-T manufacturing practices.
尽管CD19嵌合抗原受体T细胞（CAR-T）疗法在B细胞恶性肿瘤治疗中已显示出显著成效，但仍有相当一部分患者未获得长期临床缓解。这可能受个体CAR-T输注产品质量的影响。为了阐明这一点，我们将临床结果与CAR-T输注产品的特征进行了关联分析。
在这项II期研究中，23例B细胞淋巴瘤患者和1例白血病患者接受了一或两次第三代CD19定向CAR-T细胞输注（2×108/m2）。该临床试验已在clinicaltrials.gov注册：NCT03068416。我们使用靶向单细胞RNA测序和多色流式细胞术研究了个体CD19 CAR-T输注产品的转录谱。
在本研究使用的情况下，两次CAR-T输注并不比一次更好。对于CAR-T输注产品，我们发现具有高多功能性、高细胞毒性和细胞因子产生水平以及低功能失调特征的效应样CD8+ CAR-T细胞与临床反应相关。CAR-T制造过程中延长的体外扩增时间对输注产品中效应CD8+ CAR-T细胞的比例产生了负面影响。
我们确定了与反应相关的效应CD8+ CAR-T细胞的细胞内在特征，可将其用作临床结果的指标。该研究结果也为CAR-T制造实践提供了指导。
