Abstract
Nuclear pore complexes (NPCs) are sophisticated transporters assembled from diverse proteins termed nucleoporins (Nups). They control all nucleocytoplasmic transport and form a stringent barrier between the cytosol and the nucleus. While selective receptor‐mediated transport enables translocation of macromolecules up to striking sizes approaching megadalton‐scale, the upper cutoff for diffusion is at 40 kDa. Raising the cutoff is of particular importance for nuclear delivery of therapeutic nanoparticles, for example, gene and chemotherapy. In this work, we set out to present compounds capable of raising the cutoff to an extent enabling nuclear delivery of 6 kbp pDNA (150 kDa) in cultured human vascular endothelial cells. Of all tested compounds one is singled out, 1,6‐hexanediol (1,6‐HD). Our observations reveal that 1,6‐HD facilitates nuclear delivery of pDNA in up to 10–20% of the tested cells, compared to no delivery at all in control conditions. It acts by interfering with bonds between Nups that occupy the NPC channel and confer transport selectivity. It also largely maintains cell viability even at high concentrations. We envisage that 1,6‐HD may serve as a lead substance and usher in the design of potent new strategies to increase nuclear delivery of therapeutic nanoparticles.
Highlights
Nuclear pore complexes (NPCs) are proteinaceous structures that span the nuclear envelope at regular distances and form the sole pathways for nucleocytoplasmic transport
Their structural arrangement inside the NPC channel generates a barrier that determines the upper cut-off for passive diffusion of molecules, which is at a diameter of 5 nm or a size of 40 kDa.[1]
We present 1,6-hexanediol as a potent NPC barrier breaker (NBB), that facilitates nuclear delivery of pDNA with significantly higher efficacy as compared to 1,2-TCHD, while maintaining viability of cells
Summary
Nuclear pore complexes (NPCs) are proteinaceous structures that span the nuclear envelope at regular distances and form the sole pathways for nucleocytoplasmic transport. Nups with motifs rich in hydrophobic phenylalanine and glycine (FG) repeats, connected through hydrophilic spacer sequences.[2] FG-Nups are abundant inside the center of the NPC channel and are responsible for selective transport.[2] Their structural arrangement inside the NPC channel generates a barrier that determines the upper cut-off for passive diffusion of molecules, which is at a diameter of 5 nm or a size of 40 kDa.[1] Receptor-mediated transport on the other hand, is highly selective and enables the translocation of far greater molecules up to 39 nm.[3] Such molecules, for instance proteins, bear nuclear localization signals, which are recognized by soluble transport receptors The latter regularly shuttle between the cytosol and the nucleus and are. Increasing the upper cut-off limit of the FG-Nups barrier is highly beneficial for delivering diverse therapeutic nanoparticles to their place of action, the nucleus, for instance for cancer and gene therapies.[10,11] Several amphiphilic alcohols disrupt NPC selectivity and break its barrier function by acting on FG-Nups, for example, trans-1,2-cyclohexanediol (1,2-TCHD).[12,13,14,15] Upon the addition of this NPC barrier breaker (NBB), NPCs become permeable to 70 kDa dextran[16,17] and single pDNA particles.13 1,2-TCHD, exhibits significant cytotoxic effects.[13,16] In this study, we present 1,6-hexanediol as a potent NBB, that facilitates nuclear delivery of pDNA with significantly higher efficacy as compared to 1,2-TCHD, while maintaining viability of cells
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
