PO.IM01.06 · 免疫学
用IL18武装ROR1 CAR T细胞可在体外和体内改善CAR T细胞针对胰腺癌细胞系的功能和细胞毒性
Armoring ROR1 CAR T cells with IL18 improves CAR T cell function and cytotoxicity against pancreatic cancer cell-lines in vitro and in vivo
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
ROR1（受体酪氨酸激酶样孤儿受体1，Receptor tyrosine kinase-like Orphan Receptor 1）是一种在实体瘤中广泛表达的肿瘤相关抗原，是胰腺癌CAR T细胞疗法的一个有前景的靶点。基于细胞的免疫疗法迄今成功有限，部分原因在于免疫抑制性的肿瘤微环境（TME）。在此我们证明，用白细胞介素-18（IL18）武装我们此前所描述的1 ROR1 CAR T细胞产品，可在体外和体内增强其针对胰腺细胞系的扩增、持久性和抗肿瘤疗效。
利用表达第二代CAR的慢病毒载体（LV），在人T细胞中生成了ROR1特异性CAR-T细胞，分别为单独CAR、CAR+组成型IL-18或CAR+NFAT-IL18。与单独CAR相比，IL18-ROR1 CAR T细胞的扩增高出1.5倍，保留了初始（naïve）表型，并且LAG3表达降低（37% vs 81%）。通过xCelligence肿瘤共培养试验测定，武装型CAR在体外对ROR1(+)肿瘤细胞系的细胞毒性等于或大于单独CAR。在AsPC-1小鼠异种移植模型中，每种CAR构建体均表现出抗肿瘤疗效，其中表达IL-18的CAR显示出显著更强的肿瘤控制。与缺乏IL-18的ROR1 CAR-T相比，组成型IL18-CAR在T细胞输注后第28天完全清除了肿瘤，并在整个研究过程中显示出更高的持久性（图1）。与UTD相比，CAR+NFAT-IL18同样控制了肿瘤生长。
总之，IL18武装型CAR T细胞在胰腺癌异种移植模型中显著改善了ROR1 CAR的扩增、持久性和抗肿瘤疗效。重要的是，基于NFAT启动子对ROR1 CAR T进行调控可能提供一种可行的方法，在保留IL-18诱导的增强活性的同时调控潜在毒性。这一武装策略可能为实体瘤适应症提供一种有前景的方法，值得进一步开展临床前和转化研究。
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图1. 靶向胰腺细胞系的ROR1 CAR T的有效组成型和诱导型IL18武装的体内验证。* p<0.05，** p<0.01，*** p<0.001
参考文献：
1. Tran TM等. J Immunother Cancer 2024;12(4)
查看英文原文 English abstract
ROR1 ( R eceptor tyrosine kinase-like O rphan R eceptor 1 ) is a tumor-associated antigen widely expressed in solid tumors and is a promising target for CAR T cell therapy in pancreatic cancer. Cell-based immunotherapy has met with limited success due in part to the immunosuppressive tumor microenvironment (TME). Here we demonstrate that armoring our previously described 1 ROR1 CAR T cell product with interleukin-18 (IL18) enhanced expansion, persistence, and antitumor efficacy against pancreatic cell lines both in vitro and in vivo .
ROR-1-specific CAR-T cells were generated in human T cells using lentiviral vectors (LV) expressing a second generation CAR alone, CAR+ constitutive IL-18, or CAR+NFAT-IL18. IL18-ROR1 CAR T cells expanded 1.5 fold more, preserved naïve phenotype, and had reduced LAG3 expression (37% vs 81%) vs CAR. In vitro armored CAR cytotoxicity, determined by xCelligence tumor co-culture assays, was equal to or greater than CAR alone against ROR1(+) tumor cell lines. In an AsPC-1 mouse xenograft model, each CAR construct exhibited anti-tumor efficacy, with IL-18 expressing CARs displaying significantly greater tumor control. The constitutive IL18-CAR completely cleared tumor at day 28 post-T cell infusion and showed increased persistence throughout the study when compared to ROR1 CAR-T lacking IL-18 (Fig.1). CAR+NFAT-IL18 similarly controlled tumor growth compared to UTD.
In summary, IL18 armored CAR T cells significantly improved the expansion, persistence, and antitumor efficacy of ROR1 CARs in pancreatic cancer xenograft models. Importantly, NFAT-promoter-based regulation of ROR1 CAR T may offer a viable approach to regulating potential toxicity while retaining boosted IL-18 induced activity. This armoring strategy may offer a promising approach solid tumor indications and warrants further preclinical and translational investigation.
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Fig 1. In vivo demonstration of effective constitutive and inducible IL18 armoring of ROR1 CAR T targeting pancreatic cell-lines. * p<0.05, ** p<0.01, *** p<0.001
Reference:
1. Tran TM et al. J Immunother Cancer 2024;12(4)
利益披露 Disclosure
B. Chand Thakuri,
Miltenyi Biotec Employment.
S. Nurmukhambetova,
Miltenyi Biotec Employment.
T. Tran,
Miltenyi Biotec Employment.
N. Tran,
Miltenyi Biotec Employment.
P. Hu,
Miltenyi Biotec Employment.
P. Dash,
Miltenyi Biotec Employment.
R. Orentas,
Miltenyi Biotec Employment.
A. Carter,
Miltenyi biotec Employment.