Xing Yuqi, Zhang Feiyu, Ji Panpan, Wei Mengying, Yin Chunhui, Yang Angang, Yang Guodong, Zhao Jing
The State Laboratory of Cancer Biology, Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China.
Department of Cardiology, Xijing Hospital, Fourth Military Medical University, Xi'an, Shaanxi, 710032, China.
Small. 2023 May;19(20):e2204031. doi: 10.1002/smll.202204031. Epub 2023 Jan 12.
Pyroptosis is a newly discovered inflammatory form of programmed cell death, which promotes systemic immune response in cancer immunotherapy. GSDMD is one of the key molecules executing pyroptosis, while therapeutical delivery of GSDMD to tumor cells is of great challenge. In this study, an extracellular vesicles-based GSDMD-N mRNA delivery system (namely EV ) is developed for enhanced cancer immunotherapy, with GSDMD-N mRNA encapsulated inside, Ce6 (Chlorin e6 (Ce6), a hydrophilic sensitizer) incorporated into extracellular vesicular membrane, and HER2 antibody displayed onto the surface. Briefly, GSDMD-N mRNA is translationally repressed in donor cells by optimized puromycin, ensuring the cell viability and facilitating the mRNA encapsulation into extracellular vesicles. When targeted and delivered into HER2 breast cancer cells by the engineered extracellular vesicles, the translational repression is unleashed in the recipient cells as the puromycin is diluted and additionally inactivated by sonodynamic treatment as the extracellular vesicles are armed with Ce6, allowing GSDMD-N translation and pyroptosis induction. In addition, sonodynamic treatment also induces cell death in the recipient cells. In the SKBR3- and HER2 transfected 4T1- inoculated breast tumor mouse models, the engineered EV efficiently induces a powerful tumor immune response and suppressed tumor growth, providing a nanoplatform for cancer immunotherapy.
细胞焦亡是一种新发现的程序性细胞死亡的炎症形式，在癌症免疫治疗中可促进全身免疫反应。Gasdermin D（GSDMD）是执行细胞焦亡的关键分子之一，而将GSDMD递送至肿瘤细胞进行治疗具有很大挑战。在本研究中，开发了一种基于细胞外囊泡的GSDMD-N mRNA递送系统（即EV）用于增强癌症免疫治疗，其中GSDMD-N mRNA包裹在内部，Ce6（二氢卟吩e6（Ce6），一种亲水性敏化剂）掺入细胞外囊泡膜中，并在表面展示HER2抗体。简而言之，通过优化的嘌呤霉素在供体细胞中抑制GSDMD-N mRNA的翻译，确保细胞活力并促进mRNA包裹到细胞外囊泡中。当通过工程化细胞外囊泡靶向递送至HER2阳性乳腺癌细胞时，随着嘌呤霉素被稀释且细胞外囊泡携带Ce6后通过声动力处理使其额外失活，受体细胞中的翻译抑制被解除，从而允许GSDMD翻译并诱导细胞焦亡。此外，声动力处理还可诱导受体细胞死亡。在SKBR3和HER2转染的4T1接种的乳腺肿瘤小鼠模型中，工程化的EV有效诱导强大的肿瘤免疫反应并抑制肿瘤生长，为癌症免疫治疗提供了一个纳米平台。
