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Synthetic nanoparticle vehicles deliver siRNA to vascular endothelial cells for VCAM‐1 knockdown
Author(s) -
Kaneda Megan M,
Milbrandt Jeffrey D,
Lanza Gregory M,
Wickline Samuel A
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.1_supplement.902.18
Subject(s) - transfection , vcam 1 , gene knockdown , flow cytometry , small interfering rna , viability assay , chemistry , in vivo , microbiology and biotechnology , biophysics , cell , cell adhesion , biology , biochemistry , apoptosis , gene
The therapeutic promise of siRNA relies on its ability to knock down genes with minimal side effects. Upregulation of inflammatory genes such as vascular cell adhesion molecule‐1 (VCAM‐1) in atherosclerosis suggests that targeted delivery of siRNA as an antiatherosclerotic therapy might modify disease progression. We have shown previously that liquid perfluorocarbon nanoparticle emulsions (PFC‐NP) can serve as molecularly targeted vehicles for imaging and drug delivery in vivo . In this study, we sought to utilize PFC NP as carriers of siRNA to endothelial cells. Cationic PFC‐NP emulsions were produced with 1,2‐Dioleoyl‐3‐Trimethylammonium‐Propane (DOTAP). siRNA to VCAM‐1 was complexed to the emulsions and incubated with mouse 2F‐2B endothelial cells. We observed that fluorescently labeled siRNA in cells 24 h after transfection as determined by confocal microscopy. Optimized transfection conditions (10 pM PFC‐NP with 5 nM siRNA) produce a 52% decrease in VCAM‐1 mRNA levels after 48h. Corresponding viability data measured 24 h after transfection using flow cytometry indicated 80–90% viability in 2F‐2B cells under similar transfection conditions. Accordingly, these particles offer an efficient method for siRNA delivery to endothelial cells and they can simultaneously serve as a vehicle for molecular imaging to confirm siRNA delivery. This work was supported by NIH grant #HL073646