Megalin-Mediated Oligonucleotide Trafficking for Breast Cancer Chemosensitization
Abstract
The delivery of the large and charged oligonucleotides such as siRNA to cancer cells is a highly inefficient process. Megalin is an endocytotic surface receptor highly expressed in breast cancer cells that can be exploited for enhanced cellular siRNA uptake. The goals of this study are to develop the necessary tools and validate a novel strategy to target megalin for improved delivery of anticancer siRNA. Lipid nanocarriers (LNC) for delivery of fluorescent siRNA or siRNA targeting clusterin were prepared. The LNC system was shown capable of binding the megalin targeting substrate ApoE at 91.2 4.8% efficiency. The ApoE-binding on LNC was shown highly stable. Only 8.5 3.3% of the bound ApoE came off after 48 hr incubation at 37 deg C. The encapsulation efficiency of siRNA was 75.2 6.1%. After siRNA encapsulation, the size of ApoE-LNC was measured at 241 nm (PDI = 0.168). This system demonstrated very low non-specific toxicity as measured by trypan blue assay. The uptake of fluorescent LNC into MCF-7 cells was visibly increased several folds after ApoE modification, showing the value of megalin targeting for siRNA delivery. Our early data have also demonstrated strong megalin and clusterin expression in MCF-7 cells, indirectly supporting the potential value of the proposed targeted strategy. Future studies will further confirm the correlation between ApoE coating of a siRNA carrier and megalin expression and clusterin knockdown.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2010
- Accession Number
- ADA542623
Entities
People
- Ho-lun Wong
Organizations
- Temple University