Tumor delivery of antisense oligomer using trastuzumab within a streptavidin nanoparticle
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Trastuzumab (Herceptin™) is often internalized following binding to Her2+ tumor cells. The objective of this study was to investigate whether trastuzumab can be used as a specific carrier to deliver antisense oligomers into Her2+ tumor cells both in vitro and in vivo.
A biotinylated MORF oligomer antisense to RhoC mRNA and its biotinylated sense control were labeled with either lissamine for fluorescence detection or 99mTc for radioactivity detection and were linked to biotinylated trastuzumab via streptavidin. The nanoparticles were studied in SUM190 (RhoC+, Her2+) study and SUM149 (RhoC+, Her2−) control cells in culture and as xenografts in mice.
As evidence of unimpaired Her2+ binding of trastuzumab within the nanoparticle, accumulations were clearly higher in SUM190 compared to SUM149 cells and, by whole-body imaging, targeting of SUM190 tumor was similar to that expected for a radiolabeled trastuzumab. As evidence of internalization, fluorescence microscopy images of cells grown in culture and obtained from xenografts showed uniform cytoplasm distribution of the lissamine-MORF. An invasion assay showed decreased RhoC expression in SUM190 cells when incubated with the antisense MORF nanoparticles at only 100 nM.
Both in cell culture and in animals, the nanoparticle with trastuzumab as specific carrier greatly improved tumor delivery of the antisense oligomer against RhoC mRNA into tumor cells overexpressing Her2 and may be of general utility.
KeywordsAntisense Trastuzumab Streptavidin MORF oligomer Nanoparticle
The authors are grateful to Dr. Hendrik Pretorius, University of Massachusetts Medical School, for help with image quantitation, and to Dr. Kayoko Nakamura, Department of Radiology, Keio University School of Medicine, Tokyo 160-8582, Japan, for helpful discussion. This research was supported by the Congressionally Directed Medical Research Programs (CDMRP), grant No. W81XWH-06-1-0649 and in part the National Institutes of Health, grant No. SR21CA100092-02.
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