Nanomechanical action opens endo-lysosomal compartments

Abstract

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.

Rational Design of Lipid-Based Vectors for Advanced Therapeutic Vaccines

“…Current strategies used by lipid-based vectors for endosomal escape include: (a) “proton sponge effect” mediated by a high pH-buffering material that swells when protonated; (b) pH-responsive charge switching that increases the interaction with endosomal membranes; and (c) membrane fusion-mediated endosomal escape by integrating fusogenic lipids into vectors. Recent studies found that vector-topology (cuboplex nanostructure) mediated membrane fusion and that mechanical-action-induced endosomal membrane damage can also improve the endosomal escape efficiency [ 22 , 23 ].…”

“…Endosomal escape is pivotal for intracellular delivery of vaccines, which involves transporting vaccines directly into cells, to target specific cellular processes and enhance the effectiveness of treatments at the molecular level. Xu et al introduced an innovative mechanism via lipid-based nanoscale molecular machines, which utilized a light-induced nanomechanical action to destabilize endo-lysosomal compartments, enhancing the cytosolic delivery of therapeutic agents [ 23 ]. Building on this concept of improving delivery efficacy, Choi et al further advanced LNP technology by incorporating histidinamide-conjugated cholesterol into LNP formulations, thereby improving mRNA delivery by facilitating endosomal escape [ 37 ].…”