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Endosomal size and membrane leakiness influence proton sponge-based rupture of endosomal vesicles

Lotte Vermeulen (UGent) , Toon Brans (UGent) , Sangram Keshari Samal (UGent) , Peter Dubruel (UGent) , Jo Demeester (UGent) , Stefaan De Smedt (UGent) , Katrien Remaut (UGent) and Kevin Braeckmans (UGent)
(2018) ACS NANO. 12(3). p.2332-2345
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Center for nano- and biophotonics (NB-Photonics)
Abstract
In gene therapy, endosomal escape represents a major bottleneck since nanoparticles often remain entrapped inside endosomes and are trafficked toward the lysosomes for degradation. A detailed understanding of the endosomal barrier would be beneficial for developing rational strategies to improve transfection and endosomal escape. By visualizing individual endosomal escape events in live cells, we obtain insight into mechanistic factors that influence proton sponge-based endosomal escape. In a comparative study, we found that HeLa cells treated with JetPEI/pDNA polyplexes have a 3.5-fold increased endosomal escape frequency compared to ARPE-19 cells. We found that endosomal size has a major impact on the escape capacity. The smaller HeLa endosomes are more easily ruptured by the proton sponge effect than the larger ARPE-19 endosomes, a finding supported by a mathematical model based on the underlying physical principles. Still, it remains intriguing that even in the small HeLa endosomes, <10% of the polyplex-containing endosomes show endosomal escape. Further experiments revealed that the membrane of polyplex-containing endosomes becomes leaky to small compounds, preventing effective buildup of osmotic pressure, which in turn prevents endosomal rupture. Analysis of H1299 and A549 cells revealed that endosomal size determines endosomal escape efficiency when cells have comparable membrane leakiness. However, at high levels of membrane leakiness, buildup of osmotic pressure is no longer possible, regardless of endosomal size. Based on our findings that both endosomal size and membrane leakiness have a high impact on proton sponge-based endosomal rupture, we provide important clues toward further improvement of this escape strategy.
Keywords
gene therapy, nanomedicine, polyethylenimine, endosomal escape, proton sponge effect, SYNTHETIC PHOSPHOLIPID-VESICLES, MEDIATED GENE DELIVERY, NUCLEIC-ACID DELIVERY, INTRACELLULAR DELIVERY, LINEAR POLYETHYLENIMINE, CELLULAR UPTAKE, DNA COMPLEXES, HELA-CELLS, ENDOCYTOSIS, ESCAPE

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Citation

Please use this url to cite or link to this publication:

Chicago
Vermeulen, Lotte, Toon Brans, Sangram Keshari Samal, Peter Dubruel, Jo Demeester, Stefaan De Smedt, Katrien Remaut, and Kevin Braeckmans. 2018. “Endosomal Size and Membrane Leakiness Influence Proton Sponge-based Rupture of Endosomal Vesicles.” Acs Nano 12 (3): 2332–2345.
APA
Vermeulen, Lotte, Brans, T., Samal, S. K., Dubruel, P., Demeester, J., De Smedt, S., Remaut, K., et al. (2018). Endosomal size and membrane leakiness influence proton sponge-based rupture of endosomal vesicles. ACS NANO, 12(3), 2332–2345.
Vancouver
1.
Vermeulen L, Brans T, Samal SK, Dubruel P, Demeester J, De Smedt S, et al. Endosomal size and membrane leakiness influence proton sponge-based rupture of endosomal vesicles. ACS NANO. 2018;12(3):2332–45.
MLA
Vermeulen, Lotte, Toon Brans, Sangram Keshari Samal, et al. “Endosomal Size and Membrane Leakiness Influence Proton Sponge-based Rupture of Endosomal Vesicles.” ACS NANO 12.3 (2018): 2332–2345. Print.
@article{8570812,
  abstract     = {In gene therapy, endosomal escape represents a major bottleneck since nanoparticles often remain entrapped inside endosomes and are trafficked toward the lysosomes for degradation. A detailed understanding of the endosomal barrier would be beneficial for developing rational strategies to improve transfection and endosomal escape. By visualizing individual endosomal escape events in live cells, we obtain insight into mechanistic factors that influence proton sponge-based endosomal escape. In a comparative study, we found that HeLa cells treated with JetPEI/pDNA polyplexes have a 3.5-fold increased endosomal escape frequency compared to ARPE-19 cells. We found that endosomal size has a major impact on the escape capacity. The smaller HeLa endosomes are more easily ruptured by the proton sponge effect than the larger ARPE-19 endosomes, a finding supported by a mathematical model based on the underlying physical principles. Still, it remains intriguing that even in the small HeLa endosomes, {\textlangle}10\% of the polyplex-containing endosomes show endosomal escape. Further experiments revealed that the membrane of polyplex-containing endosomes becomes leaky to small compounds, preventing effective buildup of osmotic pressure, which in turn prevents endosomal rupture. Analysis of H1299 and A549 cells revealed that endosomal size determines endosomal escape efficiency when cells have comparable membrane leakiness. However, at high levels of membrane leakiness, buildup of osmotic pressure is no longer possible, regardless of endosomal size. Based on our findings that both endosomal size and membrane leakiness have a high impact on proton sponge-based endosomal rupture, we provide important clues toward further improvement of this escape strategy.},
  author       = {Vermeulen, Lotte and Brans, Toon and Samal, Sangram Keshari and Dubruel, Peter and Demeester, Jo and De Smedt, Stefaan and Remaut, Katrien and Braeckmans, Kevin},
  issn         = {1936-0851},
  journal      = {ACS NANO},
  keyword      = {gene therapy,nanomedicine,polyethylenimine,endosomal escape,proton sponge effect,SYNTHETIC PHOSPHOLIPID-VESICLES,MEDIATED GENE DELIVERY,NUCLEIC-ACID DELIVERY,INTRACELLULAR DELIVERY,LINEAR POLYETHYLENIMINE,CELLULAR UPTAKE,DNA COMPLEXES,HELA-CELLS,ENDOCYTOSIS,ESCAPE},
  language     = {eng},
  number       = {3},
  pages        = {2332--2345},
  title        = {Endosomal size and membrane leakiness influence proton sponge-based rupture of endosomal vesicles},
  url          = {http://dx.doi.org/10.1021/acsnano.7b07583},
  volume       = {12},
  year         = {2018},
}

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