Efficient transfection of blood - brain barrier endothelial cells by lipoplexes and polyplexes in the presence of nuclear targeting NLS-PEG-acridine conjugates

Hongwei Zhang, Anton Mitin, Serguei V. Vinogradov

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Brain capillary endothelial cells of the blood - brain barrier (BBB) are difficult targets for nonviral transfection even for the most potent transfection agents. Efficient protection and nuclear delivery of plasmid DNA are the key requirements for enhancing the transfection. We designed novel DNA intercalating conjugates of PEG - tris(acridine) with a short nuclear localization signal (NLS) peptide and investigated the effect of their complexes with luciferase-encoded plasmid DNA on lipoplex- and polyplex-mediated transfection of murine brain capillary endothelial bEnd.3 cells. These intercalation complexes protected DNA from nucleolytic degradation forming a protective PEG layer around plasmid DNA and could be efficiently condensed by Lipofectamine2000 or Exgen500 into nanosized particles. Complexation of plasmid DNA with a PEG-acridine/NLS-PEG-acridine mixture (9:1 w/w), taken in an amount equal to 5 - 6 NLS peptides per DNA molecule, significantly enhanced both lipo- and polyplex transfection efficacies and increased the number of transfected bEnd.3 endothelial cells in the presence of serum. Comparative transgene expression efficiency was significantly higher at longer PEG linker and optimal conjugate-to-DNA weight ratio, especially, at lower N/P ratio for both transfection agents, reaching 15-16-fold for lipoplexes and 10-11-fold for polyplexes. In addition, the NLS-PEG-acridine conjugates did not increase cytotoxicity of lipoplexes and polyplexes to bEnd.3 cells. These conjugates can serve as promising components for development of systemic nonviral transfecting approach to the transfection of the BBB and temporary modulation of its drug permeability.

Original languageEnglish (US)
Pages (from-to)120-128
Number of pages9
JournalBioconjugate Chemistry
Volume20
Issue number1
DOIs
StatePublished - Jan 1 2009

Fingerprint

Acridines
Nuclear Localization Signals
Endothelial cells
Blood-Brain Barrier
Polyethylene glycols
Transfection
DNA
Endothelial Cells
Plasmids
Protein Sorting Signals
Brain
Cytotoxicity
Intercalation
Complexation
Luciferases
Transgenes
Permeability
Modulation
Weights and Measures
Degradation

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Pharmacology
  • Pharmaceutical Science
  • Organic Chemistry

Cite this

Efficient transfection of blood - brain barrier endothelial cells by lipoplexes and polyplexes in the presence of nuclear targeting NLS-PEG-acridine conjugates. / Zhang, Hongwei; Mitin, Anton; Vinogradov, Serguei V.

In: Bioconjugate Chemistry, Vol. 20, No. 1, 01.01.2009, p. 120-128.

Research output: Contribution to journalArticle

@article{31584897eb694cf8a1e5a6dd0f068489,
title = "Efficient transfection of blood - brain barrier endothelial cells by lipoplexes and polyplexes in the presence of nuclear targeting NLS-PEG-acridine conjugates",
abstract = "Brain capillary endothelial cells of the blood - brain barrier (BBB) are difficult targets for nonviral transfection even for the most potent transfection agents. Efficient protection and nuclear delivery of plasmid DNA are the key requirements for enhancing the transfection. We designed novel DNA intercalating conjugates of PEG - tris(acridine) with a short nuclear localization signal (NLS) peptide and investigated the effect of their complexes with luciferase-encoded plasmid DNA on lipoplex- and polyplex-mediated transfection of murine brain capillary endothelial bEnd.3 cells. These intercalation complexes protected DNA from nucleolytic degradation forming a protective PEG layer around plasmid DNA and could be efficiently condensed by Lipofectamine2000 or Exgen500 into nanosized particles. Complexation of plasmid DNA with a PEG-acridine/NLS-PEG-acridine mixture (9:1 w/w), taken in an amount equal to 5 - 6 NLS peptides per DNA molecule, significantly enhanced both lipo- and polyplex transfection efficacies and increased the number of transfected bEnd.3 endothelial cells in the presence of serum. Comparative transgene expression efficiency was significantly higher at longer PEG linker and optimal conjugate-to-DNA weight ratio, especially, at lower N/P ratio for both transfection agents, reaching 15-16-fold for lipoplexes and 10-11-fold for polyplexes. In addition, the NLS-PEG-acridine conjugates did not increase cytotoxicity of lipoplexes and polyplexes to bEnd.3 cells. These conjugates can serve as promising components for development of systemic nonviral transfecting approach to the transfection of the BBB and temporary modulation of its drug permeability.",
author = "Hongwei Zhang and Anton Mitin and Vinogradov, {Serguei V.}",
year = "2009",
month = "1",
day = "1",
doi = "10.1021/bc8003414",
language = "English (US)",
volume = "20",
pages = "120--128",
journal = "Bioconjugate Chemistry",
issn = "1043-1802",
publisher = "American Chemical Society",
number = "1",

}

TY - JOUR

T1 - Efficient transfection of blood - brain barrier endothelial cells by lipoplexes and polyplexes in the presence of nuclear targeting NLS-PEG-acridine conjugates

AU - Zhang, Hongwei

AU - Mitin, Anton

AU - Vinogradov, Serguei V.

PY - 2009/1/1

Y1 - 2009/1/1

N2 - Brain capillary endothelial cells of the blood - brain barrier (BBB) are difficult targets for nonviral transfection even for the most potent transfection agents. Efficient protection and nuclear delivery of plasmid DNA are the key requirements for enhancing the transfection. We designed novel DNA intercalating conjugates of PEG - tris(acridine) with a short nuclear localization signal (NLS) peptide and investigated the effect of their complexes with luciferase-encoded plasmid DNA on lipoplex- and polyplex-mediated transfection of murine brain capillary endothelial bEnd.3 cells. These intercalation complexes protected DNA from nucleolytic degradation forming a protective PEG layer around plasmid DNA and could be efficiently condensed by Lipofectamine2000 or Exgen500 into nanosized particles. Complexation of plasmid DNA with a PEG-acridine/NLS-PEG-acridine mixture (9:1 w/w), taken in an amount equal to 5 - 6 NLS peptides per DNA molecule, significantly enhanced both lipo- and polyplex transfection efficacies and increased the number of transfected bEnd.3 endothelial cells in the presence of serum. Comparative transgene expression efficiency was significantly higher at longer PEG linker and optimal conjugate-to-DNA weight ratio, especially, at lower N/P ratio for both transfection agents, reaching 15-16-fold for lipoplexes and 10-11-fold for polyplexes. In addition, the NLS-PEG-acridine conjugates did not increase cytotoxicity of lipoplexes and polyplexes to bEnd.3 cells. These conjugates can serve as promising components for development of systemic nonviral transfecting approach to the transfection of the BBB and temporary modulation of its drug permeability.

AB - Brain capillary endothelial cells of the blood - brain barrier (BBB) are difficult targets for nonviral transfection even for the most potent transfection agents. Efficient protection and nuclear delivery of plasmid DNA are the key requirements for enhancing the transfection. We designed novel DNA intercalating conjugates of PEG - tris(acridine) with a short nuclear localization signal (NLS) peptide and investigated the effect of their complexes with luciferase-encoded plasmid DNA on lipoplex- and polyplex-mediated transfection of murine brain capillary endothelial bEnd.3 cells. These intercalation complexes protected DNA from nucleolytic degradation forming a protective PEG layer around plasmid DNA and could be efficiently condensed by Lipofectamine2000 or Exgen500 into nanosized particles. Complexation of plasmid DNA with a PEG-acridine/NLS-PEG-acridine mixture (9:1 w/w), taken in an amount equal to 5 - 6 NLS peptides per DNA molecule, significantly enhanced both lipo- and polyplex transfection efficacies and increased the number of transfected bEnd.3 endothelial cells in the presence of serum. Comparative transgene expression efficiency was significantly higher at longer PEG linker and optimal conjugate-to-DNA weight ratio, especially, at lower N/P ratio for both transfection agents, reaching 15-16-fold for lipoplexes and 10-11-fold for polyplexes. In addition, the NLS-PEG-acridine conjugates did not increase cytotoxicity of lipoplexes and polyplexes to bEnd.3 cells. These conjugates can serve as promising components for development of systemic nonviral transfecting approach to the transfection of the BBB and temporary modulation of its drug permeability.

UR - http://www.scopus.com/inward/record.url?scp=61849158680&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=61849158680&partnerID=8YFLogxK

U2 - 10.1021/bc8003414

DO - 10.1021/bc8003414

M3 - Article

C2 - 19067581

AN - SCOPUS:61849158680

VL - 20

SP - 120

EP - 128

JO - Bioconjugate Chemistry

JF - Bioconjugate Chemistry

SN - 1043-1802

IS - 1

ER -