The Possible “Proton Sponge ” Effect of Polyethylenimine (PEI) Does Not Include Change in Lysosomal pH

doi:10.1038/MT.2012.185

Home
/
Papers
/
The Possible “Proton Sponge ” Effect of Polyethylenimine (PEI) Does Not Include Change in Lysosomal pH

Journal Article•DOI•

The Possible “Proton Sponge ” Effect of Polyethylenimine (PEI) Does Not Include Change in Lysosomal pH

Rikke Vicki Benjaminsen¹, Maria Ahlm Mattebjerg¹, Jonas Rosager Henriksen¹, S. Moein Moghimi², Thomas Lars Andresen¹ - Show less +1 more•Institutions (2)

Technical University of Denmark¹, University of Copenhagen²

01 Jan 2013-Molecular Therapy (Nature Publishing Group)-Vol. 21, Iss: 1, pp 149-157

TL;DR: Measurements of lysosomal pH as a function of PEI content and correlate the results to the "proton sponge " hypothesis show that PEI does not induce change in lysoomic pH as previously suggested and quantification ofPEI concentrations inLysosomes makes it uncertain that the " proton sponge ' effect is the dominant mechanism of polyplex escape.

read less

About: This article is published in Molecular Therapy.The article was published on 2013-01-01 and is currently open access. It has received 616 citations till now. The article focuses on the topics: Polyethylenimine.

...read moreread less

Citations

PDF

Open Access

More filters

Journal Article•DOI•

Nanoparticle Uptake: The Phagocyte Problem.

[...]

Heather H. Gustafson¹, Dolly Holt-Casper¹, David W. Grainger¹, Hamidreza Ghandehari¹•Institutions (1)

University of Utah¹

01 Aug 2015-Nano Today

TL;DR: This review focuses on describing macrophage-based initiation of downstream hallmark immunological and inflammatory processes resulting from phagocyte exposure to and internalization of nanomaterials.

...read moreread less

882 citations

Journal Article•DOI•

Challenges associated with Penetration of Nanoparticles across Cell and Tissue Barriers: A Review of Current Status and Future Prospects.

[...]

Sutapa Barua¹, Samir Mitragotri¹•Institutions (1)

University of California, Santa Barbara¹

01 Apr 2014-Nano Today

TL;DR: This review focuses on the current understanding of penetration of NPs through biological barriers, andphasis is placed on transport barriers and not immunological barriers.

...read moreread less

788 citations

Journal Article•DOI•

Block Copolymer Micelles in Nanomedicine Applications.

[...]

Horacio Cabral, Kanjiro Miyata, Kensuke Osada, Kazunori Kataoka

29 Jun 2018-Chemical Reviews

TL;DR: By shaping these features, polymeric micelles have been propitious for delivering a wide range of therapeutics through effective sensing of targets in the body and adjustment of their properties in response to particular stimuli, modulating the activity of the loaded drugs at the targeted sites, even at the subcellular level.

...read moreread less

Abstract: Polymeric micelles are demonstrating high potential as nanomedicines capable of controlling the distribution and function of loaded bioactive agents in the body, effectively overcoming biological barriers, and various formulations are engaged in intensive preclinical and clinical testing. This Review focuses on polymeric micelles assembled through multimolecular interactions between block copolymers and the loaded drugs, proteins, or nucleic acids as translationable nanomedicines. The aspects involved in the design of successful micellar carriers are described in detail on the basis of the type of polymer/payload interaction, as well as the interplay of micelles with the biological interface, emphasizing on the chemistry and engineering of the block copolymers. By shaping these features, polymeric micelles have been propitious for delivering a wide range of therapeutics through effective sensing of targets in the body and adjustment of their properties in response to particular stimuli, modulating the act...

...read moreread less

782 citations

Journal Article•

Role of clathrin- and caveolae-mediated endocytosis in gene transfer mediated by lipo- and polyplexes

[...]

Joanna Rejman, Massimo Conese

01 Jan 2005-Cellular & Molecular Biology Letters

TL;DR: In this article, the authors investigated the effects of inhibitors of clathrin-mediated endocytosis (chlorpromazine and K(+) depletion) and of caveolae-mediated uptake (filipin and genistein) on internalization of FITC-poly-l-lysine-labeled DOTAP/DNA lipoplexes and PEI/DNA polyplexes by A549 pneumocytes and HeLa cells and on the transfection efficiencies of these complexes with the luciferase gene.

...read moreread less

Abstract: We investigated the effects of inhibitors of clathrin-mediated endocytosis (chlorpromazine and K(+) depletion) and of caveolae-mediated uptake (filipin and genistein) on internalization of FITC-poly-l-lysine-labeled DOTAP/DNA lipoplexes and PEI/DNA polyplexes by A549 pneumocytes and HeLa cells and on the transfection efficiencies of these complexes with the luciferase gene. Uptake of the complexes was assayed by fluorescence-activated cell sorting. Lipoplex internalization was inhibited by chlorpromazine and K(+) depletion but unaffected by filipin and genistein. In contrast, polyplex internalization was inhibited by all four inhibitors. We conclude that lipoplex uptake proceeds only by clathrin-mediated endocytosis, while polyplexes are taken up by two mechanisms, one involving caveolae and the other clathrin-coated pits. Transfection by lipoplexes was entirely abolished by blocking clathrin-mediated endocytosis, whereas inhibition of the caveolae pathway had no effect. By contrast, transfection mediated by polyplexes was completely blocked by genistein and filipin but was unaffected by inhibitors of clathrin-mediated endocytosis. Fluorescence colocalization studies with a lysosomal marker, AlexaFluor-dextran, revealed that polyplexes taken up by clathrin-mediated endocytosis are targeted to the lysosomal compartment for degradation, while the polyplexes internalized via caveolae escape this compartment, permitting efficient transfection.

...read moreread less

692 citations

Book Chapter•DOI•

Getting Across the Cell Membrane: An Overview for Small Molecules, Peptides, and Proteins

[...]

Nicole J. Yang¹, Marlon J. Hinner•Institutions (1)

Massachusetts Institute of Technology¹

01 Jan 2015-Methods of Molecular Biology

TL;DR: A broad overview of how certain molecules are thought to cross this barrier, and what kinds of approaches are being made to enhance the intracellular delivery of those that are impermeable is provided.

...read moreread less

Abstract: The ability to efficiently access cytosolic proteins is desired in both biological research and medicine. However, targeting intracellular proteins is often challenging, because to reach the cytosol, exogenous molecules must first traverse the cell membrane. This review provides a broad overview of how certain molecules are thought to cross this barrier, and what kinds of approaches are being made to enhance the intracellular delivery of those that are impermeable. We first discuss rules that govern the passive permeability of small molecules across the lipid membrane, and mechanisms of membrane transport that have evolved in nature for certain metabolites, peptides, and proteins. Then, we introduce design strategies that have emerged in the development of small molecules and peptides with improved permeability. Finally, intracellular delivery systems that have been engineered for protein payloads are surveyed. Viewpoints from varying disciplines have been brought together to provide a cohesive overview of how the membrane barrier is being overcome.

...read moreread less

541 citations

1
2
3
4
…
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
58
59
60
61
62
63
64
65
66
67
68
69
70
71
72
73
74
75
76
77
78
79
80
81
82
83
84
85
86
87
88
89
90
91
92
93
94
95
96
97
98
99
100
101
102
103
104
105
106
107
108
109
110
111
112
113
114
115
116
117
118
119
120
121
122
123
124

Collapse

References

PDF

Open Access

More filters

Journal Article•DOI•

A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine

[...]

Otmane Boussif¹, Frank Lezoualc'h, Maria Antonietta Zanta, Mojgan Mergny, Daniel Scherman, Barbara A. Demeneix, Jean-Paul Behr - Show less +3 more•Institutions (1)

Centre national de la recherche scientifique¹

01 Aug 1995-Proceedings of the National Academy of Sciences of the United States of America

TL;DR: Together, these properties make PEI a promising vector for gene therapy and an outstanding core for the design of more sophisticated devices because its efficiency relies on extensive lysosome buffering that protects DNA from nuclease degradation, and consequent lysOSomal swelling and rupture that provide an escape mechanism for the PEI/DNA particles.

...read moreread less

Abstract: Several polycations possessing substantial buffering capacity below physiological pH, such as lipopolyamines and polyamidoamine polymers, are efficient transfection agents per se--i.e., without the addition of cell targeting or membrane-disruption agents. This observation led us to test the cationic polymer polyethylenimine (PEI) for its gene-delivery potential. Indeed, every third atom of PEI is a protonable amino nitrogen atom, which makes the polymeric network an effective "proton sponge" at virtually any pH. Luciferase reporter gene transfer with this polycation into a variety of cell lines and primary cells gave results comparable to, or even better than, lipopolyamines. Cytotoxicity was low and seen only at concentrations well above those required for optimal transfection. Delivery of oligonucleotides into embryonic neurons was followed by using a fluorescent probe. Virtually all neurons showed nuclear labeling, with no toxic effects. The optimal PEI cation/anion balance for in vitro transfection is only slightly on the cationic side, which is advantageous for in vivo delivery. Indeed, intracerebral luciferase gene transfer into newborn mice gave results comparable (for a given amount of DNA) to the in vitro transfection of primary rat brain endothelial cells or chicken embryonic neurons. Together, these properties make PEI a promising vector for gene therapy and an outstanding core for the design of more sophisticated devices. Our hypothesis is that its efficiency relies on extensive lysosome buffering that protects DNA from nuclease degradation, and consequent lysosomal swelling and rupture that provide an escape mechanism for the PEI/DNA particles.

...read moreread less

6,213 citations

"The Possible “Proton Sponge ” Effec..." refers background in this paper

...reFerences 1. Boussif, O, Lezoualc’h, F, Zanta, MA, Mergny, MD, Scherman, D, Demeneix, B et al. (1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA 92: 7297–7301. 2. Mintzer, MA and Simanek, EE (2009). Nonviral vectors for gene delivery....
[...]
...reFerences 1. Boussif, O, Lezoualc’h, F, Zanta, MA, Mergny, MD, Scherman, D, Demeneix, B et al. (1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA 92: 7297–7301. 2. Mintzer, MA and Simanek, EE (2009). Nonviral vectors for gene delivery. Chem Rev 109: 259–302. 3. Zhang, Y, Satterlee, A and Huang, L (2012). In vivo gene delivery by nonviral vectors: overcoming hurdles? Mol Ther 20: 1298–1304. 4. Sawant, RR, Sriraman, SK, Navarro, G, Biswas, S, Dalvi, RA and Torchilin, VP (2012). Polyethyleneimine-lipid conjugate-based pH-sensitive micellar carrier for gene delivery. Biomaterials 33: 3942–3951. 5. Brito, L, Little, S, Langer, R and Amiji, M (2008). Poly(beta-amino ester) and cationic phospholipid-based lipopolyplexes for gene delivery and transfection in human aortic endothelial and smooth muscle cells. Biomacromolecules 9: 1179–1187. 6. Dunlap, DD, Maggi, A, Soria, MR and Monaco, L (1997). Nanoscopic structure of DNA condensed for gene delivery....
[...]
...reFerences 1. Boussif, O, Lezoualc’h, F, Zanta, MA, Mergny, MD, Scherman, D, Demeneix, B et al. (1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine....
[...]
...reFerences 1. Boussif, O, Lezoualc’h, F, Zanta, MA, Mergny, MD, Scherman, D, Demeneix, B et al. (1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA 92: 7297–7301. 2. Mintzer, MA and Simanek, EE (2009). Nonviral vectors for gene delivery. Chem Rev 109: 259–302. 3. Zhang, Y, Satterlee, A and Huang, L (2012). In vivo gene delivery by nonviral vectors: overcoming hurdles? Mol Ther 20: 1298–1304. 4. Sawant, RR, Sriraman, SK, Navarro, G, Biswas, S, Dalvi, RA and Torchilin, VP (2012). Polyethyleneimine-lipid conjugate-based pH-sensitive micellar carrier for gene delivery. Biomaterials 33: 3942–3951. 5. Brito, L, Little, S, Langer, R and Amiji, M (2008). Poly(beta-amino ester) and cationic phospholipid-based lipopolyplexes for gene delivery and transfection in human aortic endothelial and smooth muscle cells. Biomacromolecules 9: 1179–1187. 6. Dunlap, DD, Maggi, A, Soria, MR and Monaco, L (1997). Nanoscopic structure of DNA condensed for gene delivery. Nucleic Acids Res 25: 3095–3101. 7. Rejman, J, Bragonzi, A and Conese, M (2005). Role of clathrin- and caveolae-mediated endocytosis in gene transfer mediated by lipo- and polyplexes. Mol Ther 12: 468–474. 8. von Gersdorff, K, Sanders, NN, Vandenbroucke, R, De Smedt, SC, Wagner, E and Ogris, M (2006). The internalization route resulting in successful gene expression depends on both cell line and polyethylenimine polyplex type. Mol Ther 14: 745–753. 9. Rémy-Kristensen, A, Clamme, JP, Vuilleumier, C, Kuhry, JG and Mély, Y (2001). Role of endocytosis in the transfection of L929 fibroblasts by polyethylenimine/DNA complexes....
[...]
...reFerences 1. Boussif, O, Lezoualc’h, F, Zanta, MA, Mergny, MD, Scherman, D, Demeneix, B et al. (1995). A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA 92: 7297–7301. 2. Mintzer, MA and Simanek, EE (2009). Nonviral vectors for gene delivery. Chem Rev 109: 259–302. 3. Zhang, Y, Satterlee, A and Huang, L (2012). In vivo gene delivery by nonviral vectors: overcoming hurdles? Mol Ther 20: 1298–1304. 4. Sawant, RR, Sriraman, SK, Navarro, G, Biswas, S, Dalvi, RA and Torchilin, VP (2012). Polyethyleneimine-lipid conjugate-based pH-sensitive micellar carrier for gene delivery. Biomaterials 33: 3942–3951. 5. Brito, L, Little, S, Langer, R and Amiji, M (2008). Poly(beta-amino ester) and cationic phospholipid-based lipopolyplexes for gene delivery and transfection in human aortic endothelial and smooth muscle cells....
[...]

Journal Article•DOI•

Understanding biophysicochemical interactions at the nano–bio interface

[...]

Andre E. Nel¹, Lutz Mädler², Darrell Velegol³, Tian Xia¹, Eric M.V. Hoek⁴, Ponisseril Somasundaran⁵, Fred Klaessig, Vince Castranova⁶, Mike Thompson⁷ - Show less +5 more•Institutions (7)

University of California, Los Angeles¹, University of Bremen², Pennsylvania State University³, California NanoSystems Institute⁴, Columbia University⁵, National Institute for Occupational Safety and Health⁶, FEI Company⁷

14 Jun 2009-Nature Materials

TL;DR: Probing the various interfaces of nanoparticle/biological interfaces allows the development of predictive relationships between structure and activity that are determined by nanomaterial properties such as size, shape, surface chemistry, roughness and surface coatings.

...read moreread less

Abstract: Rapid growth in nanotechnology is increasing the likelihood of engineered nanomaterials coming into contact with humans and the environment. Nanoparticles interacting with proteins, membranes, cells, DNA and organelles establish a series of nanoparticle/biological interfaces that depend on colloidal forces as well as dynamic biophysicochemical interactions. These interactions lead to the formation of protein coronas, particle wrapping, intracellular uptake and biocatalytic processes that could have biocompatible or bioadverse outcomes. For their part, the biomolecules may induce phase transformations, free energy releases, restructuring and dissolution at the nanomaterial surface. Probing these various interfaces allows the development of predictive relationships between structure and activity that are determined by nanomaterial properties such as size, shape, surface chemistry, roughness and surface coatings. This knowledge is important from the perspective of safe use of nanomaterials.

...read moreread less

6,075 citations

Journal Article•DOI•

Nonviral Vectors for Gene Delivery

[...]

Meredith A. Mintzer¹, Eric E. Simanek¹•Institutions (1)

Texas A&M University¹

01 Feb 2009-Chemical Reviews

TL;DR: Two nonviral gene delivery systems using either biodegradable poly(D,Llactide-co-glycolide) (PLG) nanoparticles or cell penetrating peptide (CPP) complexes have been designed and studied using A549 human lung epithelial cells.

...read moreread less

Abstract: The development of nonviral vectors for safe and efficient gene delivery has been gaining considerable attention recently. An ideal nonviral vector must protect the gene against degradation by nuclease in the extracellular matrix, internalize the plasma membrane, escape from the endosomal compartment, unpackage the gene at some point and have no detrimental effects. In comparison to viruses, nonviral vectors are relatively easy to synthesize, less immunogenic, low in cost, and have no limitation in the size of a gene that can be delivered. Significant progress has been made in the basic science and applications of various nonviral gene delivery vectors; however, the majority of nonviral approaches are still inefficient and often toxic. To this end, two nonviral gene delivery systems using either biodegradable poly(D,Llactide-co-glycolide) (PLG) nanoparticles or cell penetrating peptide (CPP) complexes have been designed and studied using A549 human lung epithelial cells. PLG nanoparticles were optimized for gene delivery by varying particle surface chemistry using different coating materials that adsorb to the particle surface during formation. A variety of cationic coating materials were studied and compared to more conventional surfactants used for PLG nanoparticle fabrication. Nanoparticles (~200 nm) efficiently encapsulated plasmids encoding for luciferase (80-90%) and slowly released the same for two weeks. After a delay, moderate levels of gene expression appeared at day 5 for certain positively charged PLG particles and gene expression was maintained for at least two weeks. In contrast, gene expression mediated by polyethyleneimine (PEI) ended at day 5. PLG particles were also significantly less

...read moreread less

2,189 citations

Journal Article•DOI•

Exploring polyethylenimine-mediated DNA transfection and the proton sponge hypothesis.

[...]

Akin Akinc¹, Mini Thomas¹, Alexander M. Klibanov¹, Robert Langer¹•Institutions (1)

Massachusetts Institute of Technology¹

01 May 2005-Journal of Gene Medicine

TL;DR: The relatively high transfection efficiency of polyethylenimine vectors has been hypothesized to be due to their ability to avoid trafficking to degradative lysosomes, and according to the proton sponge hypothesis, the buffering capacity of PEI leads to osmotic swelling and rupture of endosome, resulting in the release of the vector into the cytoplasm.

...read moreread less

Abstract: Background The relatively high transfection efficiency of polyethylenimine (PEI) vectors has been hypothesized to be due to their ability to avoid trafficking to degradative lysosomes. According to the proton sponge hypothesis, the buffering capacity of PEI leads to osmotic swelling and rupture of endosomes, resulting in the release of the vector into the cytoplasm. Methods The mechanism of PEI-mediated DNA transfer was investigated using quantitative methods to study individual steps in the overall transfection process. In addition to transfection efficiency, the cellular uptake, local pH environment, and stability of vectors were analyzed. N-Quaternized (and therefore non-proton sponge) versions of PEI and specific cell function inhibitors were used to further probe the proton sponge hypothesis. Results Both N-quaternization and the use of bafilomycin A1 (a vacuolar proton pump inhibitor) reduced the transfection efficiency of PEI by approximately two orders of magnitude. Chloroquine, which buffers lysosomes, enhanced the transfection efficiency of N-quaternized PEIs and polylysine by 2–3-fold. In contrast, chloroquine did not improve the transfection efficiency of PEI. The measured average pH environment of PEI vectors was 6.1, indicating that they successfully avoid trafficking to acidic lysosomes. Significantly lower average pH environments were observed for permethyl-PEI (pH 5.4), perethyl-PEI (pH 5.1), and polylysine (pH 4.6) vectors. Cellular uptake levels of permethyl-PEI and perethyl-PEI vectors were found to be 20 and 90% higher, respectively, than that of parent PEI vectors, indicating that the reduction in transfection activity of the N-quaternized PEIs is due to a barrier downstream of cellular uptake. A polycation/DNA-binding affinity assessment showed that the more charge dense N-quaternized PEIs bind DNA less tightly than PEI, demonstrating that poor vector unpackaging was not responsible for the reduced transfection activity of the N-quaternized PEIs. Conclusions The results obtained are consistent with the proton sponge hypothesis and strongly suggest that the transfection activity of PEI vectors is due to their unique ability to avoid acidic lysosomes. Copyright © 2004 John Wiley & Sons, Ltd.

...read moreread less

1,311 citations

Journal Article•DOI•

Chloride Accumulation and Swelling in Endosomes Enhances DNA Transfer by Polyamine-DNA Polyplexes

[...]

N.D. Sonawane¹, C Francis Szoka¹, Alan S. Verkman¹•Institutions (1)

University of California, San Francisco¹

07 Nov 2003-Journal of Biological Chemistry

TL;DR: The results provide direct support for the proton sponge hypothesis and thus a rational basis for the design of improved non-viral vectors for gene delivery.

...read moreread less

1,051 citations