背景:最近,自然杀伤(NK)细胞成为各种实体瘤的治疗选择。然而,免疫抑制性肿瘤免疫微环境(TIME)可降低胰腺导管腺癌NK细胞的细胞毒性。肿瘤基质内的癌症相关成纤维细胞可以通过与NK细胞的细胞活性有关的失调因子来抑制免疫监视。在这里,研究了活化的胰腺星状细胞(aPSC)在三维(3D)共培养条件下对NK细胞介导的抗癌功效的影响。
方法:优化了PANC-1肿瘤球体(TS)与aPSC和NK-92细胞在胶原蛋白基质中的3D共培养,以使用微通道芯片鉴定条件培养基中发生的细胞相互作用和差异细胞因子谱。PANC-1TS和aPSC间接共培养,而NK-92细胞被允许使用对流培养基流渗入TS通道。
结果:与aPSC共培养可促进PANC-1TSs生长并抑制NK-92细胞的抗肿瘤细胞毒作用。在aPSC和NK-92细胞之间观察到细胞活性的相互抑制而不损害迁移能力。此外,发现NK-92细胞的杀伤活性降低与NK细胞中颗粒酶B表达降低有关。
结论:此处,一种基于PANC-1TS共培养的新型芯片时间模型,aPSC,和NK-92细胞被描述。该模型可用于研究NK细胞失调的详细机制,并探索未来的治疗干预措施以恢复肿瘤微环境中的NK细胞活性。
BACKGROUND: Recently, natural killer (NK) cells emerged as a treatment option for various solid tumors. However, the immunosuppressive tumor immune microenvironment (TIME) can reduce the cytotoxic ability of NK cells in pancreatic ductal adenocarcinoma. Cancer-associated fibroblasts within the tumor stroma can suppress immune surveillance by dysregulating factors involved in the cellular activity of NK cells. Herein, the effect of activated pancreatic stellate cells (aPSCs) on NK cell-mediated anticancer efficacy under three-dimensional (3D) coculture conditions was investigated.
METHODS: 3D cocultures of PANC-1 tumor spheroids (TSs) with aPSCs and NK-92 cells in a collagen matrix were optimized to identify the occurring cellular interactions and differential cytokine profiles in conditioned media using microchannel chips. PANC-1 TSs and aPSCs were indirectly cocultured, whereas NK-92 cells were allowed to infiltrate the TS channel using convective medium flow.
RESULTS: Coculture with aPSCs promoted PANC-1 TSs growth and suppressed the antitumor cytotoxic effects of NK-92 cells. Mutual inhibition of cellular activity without compromising migration ability was observed between aPSCs and NK-92 cells. Moreover, the reduced killing activity of NK-92 cells was found to be related with reduced granzyme B expression in NK cells.
CONCLUSIONS: Herein, a novel TIME-on-chip model based on the coculture of PANC-1 TSs, aPSCs, and NK-92 cells was described. This model may be useful for studying the detailed mechanisms underlying NK cells dysregulation and for exploring future therapeutic interventions to restore NK cell activity in the tumor microenvironment.