School of Life Science, Advanced Research Institute of Multidisciplinary Science, School of Medical Technology (Institute of Engineering Medicine), Key Laboratory of Molecular Medicine and Biotherapy, Key Laboratory of Medical Molecule Science and Pharmaceutics Engineering, Beijing Institute of Technology, Beijing, 100081, China.
Department of Radiation Oncology, the First Affiliated Hospital of Zhengzhou University, Erqi, Zhengzhou, 450000, China.
Adv Sci (Weinh). 2022 Aug;9(22):e2201135. doi: 10.1002/advs.202201135. Epub 2022 Jun 4.
Exosomes derived from natural killer (NK) cells (NEO) constitute promising antineoplastic nano-biologics because of their versatile functions in immune regulation. However, a significant augment of their immunomodulatory capability is an essential need to achieve clinically meaningful treatment outcomes. Light-activatable silencing NK-derived exosomes (LASNEO) are orchestrated by engineering the NEO with hydrophilic small interfering RNA (siRNA) and hydrophobic photosensitizer Ce6. Profiling of genes involved in apoptosis pathway with Western blot and RNA-seq in cells receiving NEO treatment reveals that NEO elicits effective NK cell-like cytotoxicity toward tumor cells. Meanwhile, reactive oxygen species (ROS) generation upon laser irradiation not only triggers substantial photodynamic therapy effect but also boosts M1 tumor-associated macrophages polarization and DC maturation in the tumor microenvironment (TME). In addition, ROS also accelerates the cellular entry and endosomal escape of siRNA in TME. Finally, siRNAs targeting PLK1 or PD-L1 induce robust gene silencing in cancer cells, and downregulation of PD-L1 restores the immunological surveillance of T cells in TME. Therefore, the proposed LASNEO exhibit excellent antitumor effects by conscripting multiple types of immune cells. Considering that its manufacture is quite simple and controllable, LASNEO show compelling potential for clinical translational application.
自然杀伤 (NK) 细胞衍生的外泌体 (NEO) 由于其在免疫调节中的多功能性，构成了有前途的抗肿瘤纳米生物制剂。然而，显著提高其免疫调节能力是实现临床有意义治疗效果的必要条件。通过用亲水性小干扰 RNA (siRNA) 和疏水性光敏剂 Ce6 工程化设计 NEO，可构建光活化沉默 NK 衍生外泌体 (LASNEO)。用 Western blot 和 RNA-seq 对接受 NEO 处理的细胞中参与凋亡途径的基因进行分析，结果表明 NEO 可引发针对肿瘤细胞的有效 NK 样细胞毒性。同时，激光照射时产生的活性氧 (ROS) 不仅引发显著的光动力治疗效应，还能促进肿瘤微环境 (TME) 中 M1 肿瘤相关巨噬细胞的极化和 DC 的成熟。此外，ROS 还能加速 TME 中 siRNA 的细胞内进入和内体逃逸。最后，针对 PLK1 或 PD-L1 的 siRNAs 可在癌细胞中诱导强大的基因沉默，下调 PD-L1 可恢复 T 细胞在 TME 中的免疫监视。因此，所提出的 LASNEO 通过招募多种类型的免疫细胞发挥出色的抗肿瘤作用。鉴于其制造相当简单且可控，LASNEO 在临床转化应用方面显示出巨大的潜力。
