Her2/neu small interfering RNA delivered in culture by a streptavidin nanoparticle
Authors
Liu, XinrongNakamura, Kayoko
Cheng, Dengfeng
Peng, Cong
Xiao, Nan
Liu, Yuxia
Chen, Ling
Rusckowski, Mary
Hnatowich, Donald J.
UMass Chan Affiliations
Department of Radiology, Division of Nuclear MedicineDocument Type
Journal ArticlePublication Date
2012-07-01Keywords
2',5'-Oligoadenylate SynthetaseAntibodies, Monoclonal
Antibodies, Monoclonal, Humanized
Biotinylation
Cell Line, Tumor
Drug Delivery Systems
Female
*Genes, erbB-2
Humans
Interferons
Nanoparticles
RNA, Messenger
RNA, Small Interfering
Receptor, ErbB-2
STAT1 Transcription Factor
Streptavidin
Transfection
delivery nanoparticle
Her2
siRNA
streptavidin
radiolabeled oligomer
Nanomedicine
Neoplasms
Oncology
Radiology
Therapeutics
Metadata
Show full item recordAbstract
A three-component nanoparticle consisting of biotinylated Trastuzumab antiHer2 antibody, tat transferring peptide and radiolabeled antisense oligomer, linked together through streptavidin, have shown promise in the delivery to Her2+ tumor in mice following intravenous administration and with evidence of radiotherapeutic efficacy. These results have encouraged us to consider the nanoparticle as a delivery vehicle for RNA interference therapy in which the radiolabeled antisense oligomer is replaced with an unlabeled siRNA duplex. The siRNA stability within the nanoparticle was first confirmed by incubation with RNase A. The interferon responses, that indicate off-target cytotoxicity, were evaluated by quantitative real-time RT-PCR in BT-474 (Her2+) human breast cancer cells by measuring the mRNA expression of 2', 5'-oligoadenylate synthetase (OAS1) and Stat-1, two key interferon-responsive genes. Thereafter the cytotoxicity induced by the siRNA nanoparticle was evaluated by a clonogenic survival assay in BT-474 cells while the Her2 expression of these target cells was evaluated for evidence of specific gene silencing. The siRNA within the three-component anti- Her2/neu siRNA nanoparticle was largely protected from RNase-dependent degradation and did not activate an interferon response. The nanoparticle effectively and significantly inhibited colony formation of the target cells and silenced the Her2 gene expression at 5 nM compared with the identical nanoparticle with a scrambled siRNA. Our delivery nanoparticle, with tumor targeting provided by the antibody and its accumulation without entrapment, possibly due to the transfecting peptide, delivered an siRNA duplex to the proper subcellular localization for specific and effective gene silencing in culture by what appears to be an siRNA mechanism.Source
Curr Drug Deliv. 2012 Jul;9(4):431-6. doi:10.2174/156720112801323035DOI
10.2174/156720112801323035Permanent Link to this Item
http://hdl.handle.net/20.500.14038/48002PubMed ID
22520071Related Resources
Link to Article in PubMedae974a485f413a2113503eed53cd6c53
10.2174/156720112801323035