Combined MUC1-specific nanobody-tagged PEG-polyethylenimine polyplex targeting and transcriptional targeting of tBid transgene for directed killing of MUC1 over-expressing tumour cells

JOURNAL OF CONTROLLED RELEASE

卷:156

Article

Sadeqzadeh, Elham¹  Rahbarizadeh, Fatemeh¹  Ahmadvand, Davoud²  Rasaee, Mohammad J.¹  Parhamifar, Ladan²  Moghimi, S. Moein² 

[1] Tarbiat Modares Univ, Dept Med Biotechnol, Fac Med Sci, Tehran, Iran

[2] Univ Copenhagen, Ctr Pharmaceut Nanotechnol & Nanotoxicol, DK-2100 Copenhagen O, Denmark

关键词: Cancer nanomedicines;    MUC1;    Nanobody;    Poly(ethylene glycol);    Polyethylenimine;    Transcriptional targeting;   

DOI  :  10.1016/j.jconrel.2011.06.022

来源: Elsevier

【 摘 要 】

We provide evidence for combining a single domain antibody (nanobody)-based targeting approach with transcriptional targeting as a safe way to deliver lethal transgenes to MUC1 over-expressing cancer cells. From a nanobody immune library, we have isolated an anti-DF3/Mucin1 (MUC1) nanobody with high specificity for the MUC1 antigen, which is an aberrantly glycosylated glycoprotein over-expressed in tumours of epithelial origin. The anti-MUC1 nanobody was covalently linked to the distal end of poly(ethylene glycol) 3500 (PEG3500) in PEG3500-25 kDa polyethylenimine (PEI) conjugates and the resultant macromolecular entity successfully condensed plasmids coding a transcriptionally targeted truncated-Bid (tBid) killer gene under the control of the cancer-specific MUC1 promoter. The engineered polyplexes exhibited favourable physicochemical characteristics for transfection and dramatically elevated the level of Bid/tBid expression in both MUC1 over-expressing caspase 3-deficient (MCF7 cells) and caspase 3-positive (T47D and SKBR3) tumour cell lines and, concomitantly, induced considerable cell death. Neither transgene expression nor cell death occurred when the MUC1 promoter was replaced with the CNS-specific synapsin I promoter. Since PEGylated PEI was only responsible for DNA compaction and played no significant role in direct transfection and cell killing, our attempts overcome previously reported PEI-mediated apoptotic and necrotic cell death, which is advantageous for future in vivo transcriptional targeting as this will minimize (or eliminate) nontargeted cell damage. (C) 2011 Elsevier B.V. All rights reserved.

【 授权许可】

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