Dumontel Bianca, Susa Francesca, Limongi Tania, Vighetto Veronica, Debellis Doriana, Canta Marta, Cauda Valentina
Department of Applied Science and Technology, Politecnico di Torino, Turin, Italy.
Electron Microscopy Facility, Istituto Italiano di Tecnologia (IIT), Genoa, Italy.
Cell Biosci. 2022 May 14;12(1):61. doi: 10.1186/s13578-022-00784-9.
We propose an efficient method to modify B-cell derived EVs by loading them with a nanotherapeutic stimuli-responsive cargo and equipping them with antibodies for efficient targeting of lymphoma cells.
The post-isolation engineering of the EVs is accomplished by a freeze-thaw method to load therapeutically-active zinc oxide nanocrystals (ZnO NCs), obtaining the so-called TrojanNanoHorse (TNH) to recall the biomimetism and cytotoxic potential of this novel nanoconstruct. TNHs are further modified at their surface with anti-CD20 monoclonal antibodies (TNH) achieving specific targeting against lymphoid cancer cell line. The in vitro characterization is carried out on CD20+ lymphoid Daudi cell line, CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (B lymphocytes). The TNH shows nanosized structure, high colloidal stability, even over time, and good hemocompatibility. The in vitro characterization shows the high biocompatibility, targeting specificity and cytotoxic capability. Importantly, the selectivity of TNH demonstrates significantly higher interaction towards the target lymphoid Daudi cell line compared to the CD20-negative cancerous myeloid cells (HL60) and the healthy counterpart (lymphocytes). An enhanced cytotoxicity directed against Daudi cancer cells is demonstrated after the TNH activation with high-energy ultrasound shock-waves (SW).
This work demonstrates the efficient re-engineering of EVs, derived from healthy cells, with inorganic nanoparticles and monoclonal antibodies. The obtained hybrid nanoconstructs can be on-demand activated by an external stimulation, here acoustic pressure waves, to exploit a cytotoxic effect conveyed by the ZnO NCs cargo against selected cancer cells.
我们提出了一种有效的方法来修饰B细胞衍生的细胞外囊泡（EVs），即向其装载纳米治疗性刺激响应性货物，并为其配备抗体以有效靶向淋巴瘤细胞。
通过冻融法完成细胞外囊泡的分离后工程，以装载具有治疗活性的氧化锌纳米晶体（ZnO NCs），从而获得所谓的特洛伊纳米马（TNH），以唤起这种新型纳米结构的仿生学和细胞毒性潜力。TNHs在其表面进一步用抗CD20单克隆抗体修饰（TNH），实现对淋巴癌细胞系的特异性靶向。对CD20 + 淋巴Daudi细胞系、CD20阴性癌性髓样细胞（HL60）和健康对照（B淋巴细胞）进行了体外表征。TNH显示出纳米尺寸结构、高胶体稳定性（即使随时间推移）以及良好的血液相容性。体外表征显示出高生物相容性、靶向特异性和细胞毒性能力。重要的是，与CD20阴性癌性髓样细胞（HL60）和健康对照（淋巴细胞）相比，TNH的选择性显示出对目标淋巴Daudi细胞系的相互作用明显更高。在用高能超声冲击波（SW）激活TNH后，对Daudi癌细胞的细胞毒性增强。
这项工作证明了用无机纳米颗粒和单克隆抗体对源自健康细胞的细胞外囊泡进行有效的重新工程改造。所获得的混合纳米结构可以通过外部刺激（此处为声压波）按需激活，以利用ZnO NCs货物对选定癌细胞传递的细胞毒性作用。
