Department of Biomedical Engineering, Tufts University, Medford, MA, 02155, USA.
Nat Commun. 2023 Oct 20;14(1):6645. doi: 10.1038/s41467-023-42280-9.
Endo-lysosomal escape is a highly inefficient process, which is a bottleneck for intracellular delivery of biologics, including proteins and nucleic acids. Herein, we demonstrate the design of a lipid-based nanoscale molecular machine, which achieves efficient cytosolic transport of biologics by destabilizing endo-lysosomal compartments through nanomechanical action upon light irradiation. We fabricate lipid-based nanoscale molecular machines, which are designed to perform mechanical movement by consuming photons, by co-assembling azobenzene lipidoids with helper lipids. We show that lipid-based nanoscale molecular machines adhere onto the endo-lysosomal membrane after entering cells. We demonstrate that continuous rotation-inversion movement of Azo lipidoids triggered by ultraviolet/visible irradiation results in the destabilization of the membranes, thereby transporting cargoes, such as mRNAs and Cre proteins, to the cytoplasm. We find that the efficiency of cytosolic transport is improved about 2.1-fold, compared to conventional intracellular delivery systems. Finally, we show that lipid-based nanoscale molecular machines are competent for cytosolic transport of tumour antigens into dendritic cells, which induce robust antitumour activity in a melanoma mouse model.
内体-溶酶体逃逸是一个效率非常低的过程，这是生物体内输送的一个瓶颈，包括蛋白质和核酸。本文中，我们展示了一种基于脂质的纳米级分子机器的设计，该机器通过在光照射下通过纳米机械作用破坏内体-溶酶体隔室，从而实现生物的有效细胞质运输。我们通过将偶氮苯脂质体与辅助脂质共组装来制造基于脂质的纳米级分子机器，该机器通过消耗光子来进行机械运动。我们表明，基于脂质的纳米级分子机器在进入细胞后会黏附在内体-溶酶体膜上。我们表明，紫外/可见照射引发的 Azo 脂质体的连续旋转-反转运动导致膜的不稳定，从而将货物（如 mRNA 和 Cre 蛋白）运送到细胞质中。我们发现，与传统的细胞内输送系统相比，细胞质输送的效率提高了约 2.1 倍。最后，我们表明，基于脂质的纳米级分子机器能够将肿瘤抗原输送到树突状细胞的细胞质中，从而在黑色素瘤小鼠模型中诱导出强烈的抗肿瘤活性。
