Faculty of Health Sciences, University of Macau, Taipa, Macao SAR, China.
Faculty of Health Sciences, University of Macau, Taipa, Macao SAR, China; Ministry of Education Frontiers Science Center for Precision Oncology, University of Macau, Taipa, Macao SAR, China.
Biosens Bioelectron. 2022 Nov 15;216:114616. doi: 10.1016/j.bios.2022.114616. Epub 2022 Aug 6.
Tumor immunotherapy has been an important advancement in cancer treatment in recent years. Compared with T cell-based therapy, natural killer (NK) cell-based therapy does not require human leukocyte antigen matching and has fewer side effects; thus, NK cell therapy has gradually attracted the attention of researchers and clinicians. Reliable and effective animal models are essential for evaluating the effects of NK cell therapy. NK cells kill cancer cells mainly through apoptosis. In this study, we first established a 3D coculture model using fluorescence resonance energy transfer (FRET)-based lung or breast cancer cells and tdTomato-labeled NK cells. We observed that cancer cells changed from green to blue when undergoing apoptosis induced by red NK cells. We then coinjected these green cancer cells with red NK cells into zebrafish to visualize the interaction between them and the killing process of NK cells against cancer cells in real-time and at single-cell resolution in circulation. Using this model, we found that NK cells can quickly kill cancer cells in zebrafish circulation in 40 min and the caspase-3 can be activated in 5-10 min. This FRET-based zebrafish tumor model can serve as a powerful in vivo tool that can facilitate the development of NK cell-based therapy. More importantly, cancer cells from cancer patients can be labeled with our apoptotic biosensor and then transplanted into zebrafish to evaluate the sensitivity of the cancer cells to NK cells to help clinicians make treatment plans that can benefit patients.
肿瘤免疫疗法近年来已成为癌症治疗的重要进展。与基于 T 细胞的疗法相比，自然杀伤 (NK) 细胞疗法不需要人类白细胞抗原匹配，且副作用较少；因此，NK 细胞疗法逐渐引起了研究人员和临床医生的关注。可靠和有效的动物模型对于评估 NK 细胞疗法的效果至关重要。NK 细胞主要通过细胞凋亡杀死癌细胞。在本研究中，我们首先使用基于荧光共振能量转移 (FRET) 的肺癌或乳腺癌细胞和 tdTomato 标记的 NK 细胞建立了 3D 共培养模型。我们观察到，当红色 NK 细胞诱导癌细胞凋亡时，癌细胞从绿色变为蓝色。然后，我们将这些绿色癌细胞与红色 NK 细胞共同注射到斑马鱼中，以实时、单细胞分辨率可视化它们之间的相互作用以及 NK 细胞在循环中对癌细胞的杀伤过程。使用这种模型，我们发现 NK 细胞可以在 40 分钟内在斑马鱼循环中快速杀死癌细胞，并且 caspase-3 可以在 5-10 分钟内被激活。这种基于 FRET 的斑马鱼肿瘤模型可以作为一种强大的体内工具，有助于开发基于 NK 细胞的疗法。更重要的是，来自癌症患者的癌细胞可以用我们的凋亡生物传感器进行标记，然后移植到斑马鱼中，以评估癌细胞对 NK 细胞的敏感性，从而帮助临床医生制定对患者有益的治疗计划。
