RT Journal Article
T1 New insights on the mechanism of polyethylenimine transfection and their implications on gene therapy and DNA vaccines
A1 Sabín Fernández, Juan Daniel
A1 Alatorre Meda, Manuel
A1 Miñones Conde, José
A1 Domínguez Arca, Vicente
A1 Prieto, Gerardo
K1 Polyethylenimine
K1 Pore formation
K1 Mimicking nuclear membranes
K1 DNA vectors
K1 Molecular dynamics simulations
AB Polyethylenimine (PEI) has been demonstrated as an efficient DNA delivery vehicle both in vitro and in vivo. There is a consensus that PEI-DNA complexes enter the cells by endocytosis and escape from endosomes by the so-called “proton sponge” effect. However, little is known on how and where the polyplexes are de-complexed for DNA transcription and replication to occur inside the cell nucleus. To better understand this issue, we (i) tracked the cell internalization of PEI upon transfection to human epithelial cells and (ii) studied the interaction of PEI with phospholipidic layers mimicking nuclear membranes. Both the biological and physicochemical experiments provided evidence of a strong binding affinity between PEI and the lipidic bilayer. Firstly, confocal microscopy revealed that PEI alone could not penetrate the cell nucleus; instead, it arranged throughout the cytoplasm and formed a sort of aureole surrounding the nuclei periphery. Secondly, surface tension measurements, fluorescence dye leakage assays, and differential scanning calorimetry demonstrated that a combination of hydrophobic and electrostatic interactions between PEI and the phospholipidic monolayers/bilayers led to the formation of stable defects along the model membranes, allowing the intercalation of PEI through the monolayer/bilayer structure. Results are also supported by molecular dynamics simulation of the pore formation in PEI-lipidic bilayers. As discussed throughout the text, these results might shed light on a the mechanism in which the interaction between PEI and the nucleus membrane might play an active role on the DNA release: on the one hand, the PEI-membrane interaction is anticipated to facilitate the DNA disassembly from the polyplex by establishing a competition with DNA for the PEI binding and on the other hand, the forming defects are expected to serve as channels for the entrance of de-complexed DNA into the cell nucleus. A better understanding of the mechanism of transfection of cationic polymers opens paths to development of more efficiency vectors to improve gene therapy treatment and the new generation of DNA vaccines
PB Elsevier
YR 2022
FD 2022
LK http://hdl.handle.net/10347/27689
UL http://hdl.handle.net/10347/27689
LA eng
NO Colloids and Surfaces B: Biointerfaces 210 (2022) 112219. https://doi.org/10.1016/j.colsurfb.2021.112219
NO This work was supported by the Spanish "Ministerio de Ciencia, Innovación y Universidades" (Project PID2019–109517RB-I00)
DS Minerva
RD 30 abr 2026