RNA-seq data and immunohistochemical analysis of patient samples were used to explore CD70 expression in CRC and PDAC patients. In addition, CD70-targeting CAR NK cells were developed to assess cytotoxic activity against CD70+ tumor cells and CAFs, and the effect of cytokine stimulation on their efficacy was evaluated. The in vitro functionality of CD70-CAR NK cells was investigated against a panel of tumor and CAF cell lines with varying CD70 expression. Lymphoma-bearing mice were used to validate in vivo potency of CD70-CAR NK cells. Lastly, to consider patient variability, CD70-CAR NK cells were tested on patient-derived organoids containing CAFs.
In this study, we identified CD70 as a target for tumor cells and CAFs in CRC and PDAC patients. Functional evaluation of CD70-directed CAR NK cells indicated that IL-15 stimulation is essential to obtain effective elimination of CD70+ tumor cells and CAFs, and to improve tumor burden and survival of mice bearing CD70+ tumors. Mechanistically, IL-15 stimulation resulted in improved potency of CD70-CAR NK cells by upregulating CAR expression and increasing secretion of pro-inflammatory cytokines, in a mainly autocrine or intracellular manner.
We disclose CD70 as an attractive target both in hematological and solid tumors. IL-15 armored CAR NK cells act as potent effectors to eliminate these CD70+ cells. They can target both tumor cells and CAFs in patients with CRC and PDAC, and potentially other desmoplastic solid tumors.
方法:使用患者样本的RNA-seq数据和免疫组织化学分析来探索CRC和PDAC患者中CD70的表达。此外,开发了靶向CD70的CARNK细胞来评估针对CD70+肿瘤细胞和CAFs的细胞毒活性。并评价细胞因子刺激对其疗效的影响。针对一组具有不同CD70表达的肿瘤和CAF细胞系研究了CD70-CARNK细胞的体外功能性。携带淋巴瘤的小鼠用于验证CD70-CARNK细胞的体内效力。最后,为了考虑患者的变异性,在含有CAF的患者来源的类器官上测试CD70-CARNK细胞。
结果:在这项研究中,我们确定CD70是CRC和PDAC患者肿瘤细胞和CAFs的靶点.CD70导向的CARNK细胞的功能评估表明,IL-15刺激对于获得CD70+肿瘤细胞和CAFs的有效消除至关重要。并改善携带CD70+肿瘤的小鼠的肿瘤负荷和存活率。机械上,IL-15刺激通过上调CAR表达和增加促炎细胞因子的分泌,导致CD70-CARNK细胞的效力提高。主要以自分泌或细胞内的方式。
结论:我们公开了CD70作为血液肿瘤和实体瘤中的有吸引力的靶标。IL-15装甲的CARNK细胞作为有效的效应子消除这些CD70+细胞。它们可以靶向CRC和PDAC患者的肿瘤细胞和CAFs,和其他潜在的促纤维化实体瘤。