Facilitating plasmid nuclear delivery by interfering with the selective nuclear pore barrier

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.

Five‐to‐Seven Carbon Glycols Severely Impair Bioenergetics and Metabolism of Aggressive Lung Cancer Cells

“…The efficacy of 1,7-Hept seems to be the strongest of all three compounds, but 1,6-HD has additional advantages as it acts on another critical target, fundamental for the survival of cancer cells, namely, nuclear pore complexes, as demonstrated in our recent studies. [20,39] Furthermore, the molecular structure of 1,5-PD, 1,6-HD, and 1,7-Hept follows the "Lipinski rule of five" for drug design [40] (Table S1, Supporting Information), which may qualify them as promising molecules for clinical applications.…”

“…The efficacy of 1,7‐Hept seems to be the strongest of all three compounds, but 1,6‐HD has additional advantages as it acts on another critical target, fundamental for the survival of cancer cells, namely, nuclear pore complexes, as demonstrated in our recent studies. [ 20,39 ]…”

Impact of the Nuclear Envelope on Malignant Transformation, Motility, and Survival of Lung Cancer Cells

“…At concentrations applied in the present work, the utilized compounds do not dissociate FG‐Nups from the NPC barrier but interfere with their mutual bonds and consequently relax the barrier, as reported in our recent publications. [ 48 , 49 ] We used time‐lapse video microscopy to observe highly metastatic A549_3R cells over a time interval of 10 h whereby they were exposed to 2% (m/v) of either NPC barrier breakers 1,2‐trans‐Cyclohexanediol (1,2‐TCHD), 1,6‐Hexanediol (1,6‐HD), in combination (1% each), or their negative controls 1,4‐Cyclohexanediol (1,4‐CHD) and 1,2,3‐Hexanetriol (1,2,3‐HT), respectively.…”

“…We know from our previous publications that the NPC barrier breakers utilized in the present work at specified concentrations, do not dissociate FG‐Nups from the NPC barrier but rather disrupt their crucial interaction with each other and consequently prevent their selective barrier function. [ 48 , 49 ] How can interference with the NPC barrier have such profound impact on the overall structural and dynamical properties of cells? The NPCs are turnstiles of nucleocytoplasmic crosstalk.…”

1,6-Hexanediol regulates angiogenesis via suppression of cyclin A1-mediated endothelial function

“…1,6-HD, an aliphatic alcohol, specifically disrupts multivalent hydrophobic interactions between protein-RNA or protein-protein that are favorable to the formation of LLPS [10,19,20]. It has been reported that 1,6-HD affects cell viability in HeLa cells while the cytotoxicity of 1,6-HD on EA.hy926 cells is minimal [21,22]. Therefore, it is plausible to reason 1,6-HD may have function in endothelial cells.…”