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Abstract：
The clinical efficacy of CAR-NK cells against CD19-expressing blood cancers has been demonstrated, and they have shown potential for treating solid tumors as well. However, the efficacy of CAR-NK cells for treating human oral tongue squamous cell carcinoma (OTSCC) has not been examined.
We assessed MUC1 expression in human OTSCC tissue and a cell line using immunohistochemistry and immunofluorescence. We constructed NK cells that express CAR targeted to MUC1 from pluripotent stem cells (iPSC-derived MUC1-targeted CAR-NK cells) and evaluated their effectiveness against OTSCC in vitro using the xCELLigence Real-Time Cell Analysis system and CCK8 assay, and in vivo by measuring xenograft growth daily in BNDG mice treated with MUC1-targeted CAR-NK cells. As controls, we used iPSC-derived NK cells and NK-free media, which were CAR-free and blank, respectively.
MUC1 expression was detected in 79.5% (66/83) of all OTSCC patients and 72.7% (24/33) of stage III and IV. In stage III and IV MUC1 positive OTSCC, 63.6% (21/33) and 48.5% (16/33) patients had a MUC1-positive cancer cell rate of more than 50% and 80%, respectively. The iPSC-derived MUC1-targeted CAR-NK cells exhibited significant cytotoxicity against MUC1-expressing OTSCC cells in vitro, in a time- and dose-dependent manner, and showed a significant inhibitory effect on xenograft growth compared to both the iPSC-derived NK cells and the blank controls. We observed no weight loss, severe hematological toxicity or NK cell-mediated death in the BNDG mice.
The MUC1-targeted CAR-NK cells had significant efficacy against human OTSCC, and their promising therapeutic response warrants further clinical trials.
We assessed MUC1 expression in human OTSCC tissue and a cell line using immunohistochemistry and immunofluorescence. We constructed NK cells that express CAR targeted to MUC1 from pluripotent stem cells (iPSC-derived MUC1-targeted CAR-NK cells) and evaluated their effectiveness against OTSCC in vitro using the xCELLigence Real-Time Cell Analysis system and CCK8 assay, and in vivo by measuring xenograft growth daily in BNDG mice treated with MUC1-targeted CAR-NK cells. As controls, we used iPSC-derived NK cells and NK-free media, which were CAR-free and blank, respectively.
MUC1 expression was detected in 79.5% (66/83) of all OTSCC patients and 72.7% (24/33) of stage III and IV. In stage III and IV MUC1 positive OTSCC, 63.6% (21/33) and 48.5% (16/33) patients had a MUC1-positive cancer cell rate of more than 50% and 80%, respectively. The iPSC-derived MUC1-targeted CAR-NK cells exhibited significant cytotoxicity against MUC1-expressing OTSCC cells in vitro, in a time- and dose-dependent manner, and showed a significant inhibitory effect on xenograft growth compared to both the iPSC-derived NK cells and the blank controls. We observed no weight loss, severe hematological toxicity or NK cell-mediated death in the BNDG mice.
The MUC1-targeted CAR-NK cells had significant efficacy against human OTSCC, and their promising therapeutic response warrants further clinical trials.
摘要：
■已经证明了CAR-NK细胞对表达CD19的血癌的临床功效，它们也显示出治疗实体瘤的潜力。然而,尚未研究CAR-NK细胞治疗人口腔舌鳞状细胞癌(OTSCC)的疗效.
■我们使用免疫组织化学和免疫荧光评估了人OTSCC组织和细胞系中的MUC1表达。我们构建了从多能干细胞（iPSC衍生的MUC1靶向CAR-NK细胞）表达靶向MUC1的CAR的NK细胞，并使用xCELLigence实时细胞分析系统和CCK8测定在体外评估了它们对OTSCC的有效性。并且通过在用MUC1靶向的CAR-NK细胞处理的BNDG小鼠中每天测量异种移植物生长来在体内进行。作为控制，我们使用iPSC衍生的NK细胞和无NK培养基，它们是无汽车和空白的，分别。
■MUC1表达在所有OTSCC患者的79.5％（66/83）和III期和IV期的72.7％（24/33）中检测到。在III和IV阶段MUC1阳性OTSCC中，63.6%(21/33)和48.5%(16/33)的患者MUC1阳性癌细胞率超过50%和80%,分别。iPSC来源的MUC1靶向CAR-NK细胞在体外对表达MUC1的OTSCC细胞表现出显著的细胞毒性,以时间和剂量依赖的方式，与iPSC来源的NK细胞和空白对照相比，对异种移植物生长均显示出明显的抑制作用。我们没有观察到体重减轻，BNDG小鼠的严重血液学毒性或NK细胞介导的死亡。
■MUC1靶向的CAR-NK细胞对人类OTSCC具有显著功效，他们有希望的治疗反应需要进一步的临床试验。
■我们使用免疫组织化学和免疫荧光评估了人OTSCC组织和细胞系中的MUC1表达。我们构建了从多能干细胞（iPSC衍生的MUC1靶向CAR-NK细胞）表达靶向MUC1的CAR的NK细胞，并使用xCELLigence实时细胞分析系统和CCK8测定在体外评估了它们对OTSCC的有效性。并且通过在用MUC1靶向的CAR-NK细胞处理的BNDG小鼠中每天测量异种移植物生长来在体内进行。作为控制，我们使用iPSC衍生的NK细胞和无NK培养基，它们是无汽车和空白的，分别。
■MUC1表达在所有OTSCC患者的79.5％（66/83）和III期和IV期的72.7％（24/33）中检测到。在III和IV阶段MUC1阳性OTSCC中，63.6%(21/33)和48.5%(16/33)的患者MUC1阳性癌细胞率超过50%和80%,分别。iPSC来源的MUC1靶向CAR-NK细胞在体外对表达MUC1的OTSCC细胞表现出显著的细胞毒性,以时间和剂量依赖的方式，与iPSC来源的NK细胞和空白对照相比，对异种移植物生长均显示出明显的抑制作用。我们没有观察到体重减轻，BNDG小鼠的严重血液学毒性或NK细胞介导的死亡。
■MUC1靶向的CAR-NK细胞对人类OTSCC具有显著功效，他们有希望的治疗反应需要进一步的临床试验。
