Efficient Gene Transfer to Human Endothelial Cells Using DNA Complexed to Adenovirus Particles

Edgell, Cora‐Jean S.; Curiel, David T.; Hu, Ping-Chuan; Marr, Henry S.

doi:10.2144/98252gt01

Cited by 20 publications

(16 citation statements)

References 12 publications

(9 reference statements)

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…To analyze the mechanisms underlying the angiogenesis and vascular stability mediated by RAMP2, we next generated EC lines that stably overexpressed RAMP2 (RAMP2O/ E cells), utilizing EAhy926 ECs ( Figure 4, A and B; see Methods for details). EAhy926 ECs are known to form capillary-like tubes on Matrigel (34,35). We observed that RAMP2O/E cells showed much greater capillary formation than did control cells in Matrigel assays (Figure 4, C-E), clearly demonstrating that upregulation of the AM-RAMP2 system enhances angiogenesis.…”

Section: Introductionmentioning

confidence: 71%

The GPCR modulator protein RAMP2 is essential for angiogenesis and vascular integrity

Ichikawa-Shindo¹,

Sakurai²,

Kamiyoshi³

et al. 2008

J. Clin. Invest.

169

159

View full text Add to dashboard Cite

Adrenomedullin (AM) is a peptide involved both in the pathogenesis of cardiovascular diseases and in circulatory homeostasis. The high-affinity AM receptor is composed of receptor activity-modifying protein 2 or 3 (RAMP2 or -3) and the GPCR calcitonin receptor-like receptor. Testing our hypothesis that RAMP2 is a key determinant of the effects of AM on the vasculature, we generated and analyzed mice lacking RAMP2. Similar to AM -/-embryos, RAMP2 -/-embryos died in utero at midgestation due to vascular fragility that led to severe edema and hemorrhage. Vascular ECs in RAMP2 -/-embryos were severely deformed and detached from the basement membrane. In addition, the abnormally thin arterial walls of these mice had a severe disruption of their typically multilayer structure. Expression of tight junction, adherence junction, and basement membrane molecules by ECs was diminished in RAMP2 -/-embryos, leading to paracellular leakage and likely contributing to the severe edema observed. In adult RAMP2 +/-mice, reduced RAMP2 expression led to vascular hyperpermeability and impaired neovascularization. Conversely, ECs overexpressing RAMP2 had enhanced capillary formation, firmer tight junctions, and reduced vascular permeability. Our findings in human cells and in mice demonstrate that RAMP2 is a key determinant of the effects of AM on the vasculature and is essential for angiogenesis and vascular integrity.

show abstract

Section: Introductionmentioning

confidence: 71%

The GPCR modulator protein RAMP2 is essential for angiogenesis and vascular integrity

Ichikawa-Shindo¹,

Sakurai²,

Kamiyoshi³

et al. 2008

J. Clin. Invest.

169

159

View full text Add to dashboard Cite

show abstract

“…A level of 20% transfection of actively dividing, subconfluent cells can be reached by protecting DNA from cytoplasmic DNAses with histone 3,22 . Together these studies indicate that even with endosome escape facilitated by adenovirus 20,21 , transport of cytoplasmic plasmid to the nucleus remains an important limiting step in nonviral gene transfer to nondividing cells.…”

mentioning

confidence: 92%

“…During lipofection, internalization of DNA-lipid complexes is efficient (>10,000 plasmids per cell) [15][16][17] , and addition of endosome disruptors such as chloroquine and dioleoyl phosphatidyl ethanolamine (DOPE) can increase expression 3,16,18,19 . However, adding replication-deficient adenovirus to lipid-DNA complexes during transfection of subconfluent human or bovine endothelial cells provides only a modest increase in transfection efficiency, from a few percent to a level of 20-35% 20,21 . A level of 20% transfection of actively dividing, subconfluent cells can be reached by protecting DNA from cytoplasmic DNAses with histone 3,22 .…”

mentioning

confidence: 99%

Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cells

1999

View full text Add to dashboard Cite

Lipofection of nondividing cells is inefficient because much of the transfected DNA is retained in endosomes, and that which escapes to the cytoplasm enters the nucleus at low rates. To improve the final rate-limiting step of nuclear import, we conjugated a nonclassical nuclear localization signal (NLS) containing the M9 sequence of heterogeneous nuclear ribonucleoprotein (hnRNP) A1, to a cationic peptide scaffold derived from a scrambled sequence of the SV40 T-antigen consensus NLS (ScT). The ScT was added to improve DNA binding of the M9 sequence. Lipofection of confluent endothelium with plasmid complexed with the M9-ScT conjugate resulted in 83% transfection and a 63-fold increase in marker gene expression. The M9-ScT conjugate localized fluorescent plasmid into the nucleus of permeabilized cells, and addition of the nuclear pore blocker wheat germ agglutinin prevented nuclear import. This method of gene transfer may lead to viral- and lipid-free transfection of nondividing cells.

show abstract

“…Human umbilical vein endothelial cells (HUVEC), which are well known to be resistant to transfection of pDNA, have been also transfected by this method [102], [103], enhancing expression of luciferase gene by a factor of 360 compared to various conventional transfection systems [104]. Conversely, toxicity increased only by a factor of 1.6 evidencing the potentiality of magnetofection.…”

Section: ) Magnetic Nanoparticlesmentioning

confidence: 99%

Nanoparticles as Nonviral Gene Delivery Vectors

Ragusa

Garcı́a

Penadés

2007

IEEE Trans.on Nanobioscience

View full text Add to dashboard Cite

Gene therapy, as therapeutic treatment to genetic or acquired diseases, is attracting much interest in the research community, leading to noteworthy developments over the past two decades. Although this field is still dominated by viral vectors, nonviral vectors have recently received an ever increasing attention in order to overcome the safety problems of their viral counterpart. This review presents the biological aspects involved in the gene delivery process and explores the recent developments and achievements of nonviral gene carriers.

show abstract

Efficient Gene Transfer to Human Endothelial Cells Using DNA Complexed to Adenovirus Particles

Cited by 20 publications

References 12 publications

The GPCR modulator protein RAMP2 is essential for angiogenesis and vascular integrity

The GPCR modulator protein RAMP2 is essential for angiogenesis and vascular integrity

Nuclear targeting peptide scaffolds for lipofection of nondividing mammalian cells

Nanoparticles as Nonviral Gene Delivery Vectors

Contact Info

Product

Resources

About