Abstract
With the increased potential of RNA interference (RNAi) as a therapeutic strategy, new noninvasive methods for detection of siRNA delivery and silencing are urgently needed. Here we describe the development of dual-purpose probes for in vivo transfer of siRNA and the simultaneous imaging of its accumulation in tumors by high-resolution magnetic resonance imaging (MRI) and near-infrared in vivo optical imaging (NIRF). These probes consisted of magnetic nanoparticles labeled with a near-infrared dye and covalently linked to siRNA molecules specific for model or therapeutic targets. Additionally, these nanoparticles were modified with a membrane translocation peptide for intracellular delivery. We show the feasibility of in vivo tracking of tumor uptake of these probes by MRI and optical imaging in two separate tumor models. We also used proof-of-principle optical imaging to corroborate the efficiency of the silencing process. These studies represent the first step toward the advancement of siRNA delivery and imaging strategies, essential for cancer therapeutic product development and optimization.
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Acknowledgements
The authors thank J. Moore and P. Pantazopoulos for technical support with animal surgery. Confocal microscopy was performed at the Confocal Microscopy Core at MGH with technical assistance from I.A.Bagayev.
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A.M. supervised the project; Z.M. conducted the bioassay, histology and cell biology experiments, and performed the in vivo MRI experiments and the in vivo and ex vivo optical imaging experiments; W.P. performed probe synthesis and characterization of the probe; C.F. performed the ex vivo MRI experiments; V.P. conducted the RT-PCR experiments. A.M., Z.M. and W.P. wrote the manuscript.
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Supplementary information
Supplementary Fig. 1
Contribution of the MPAP membrane translocation peptide to cellular uptake. (PDF 470 kb)
Supplementary Fig. 2
Biodistribution of MN-NIRF-siGFP probe 24 hrs after intravenous administration to tumor bearing mice. (PDF 462 kb)
Supplementary Fig. 3
Tumoral accumulation of MN-NIRF-siGFP probe. (PDF 259 kb)
Supplementary Fig. 4
Immunostimulatory and cytotoxic effects of MN-NIRF-siGFP. (PDF 199 kb)
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Medarova, Z., Pham, W., Farrar, C. et al. In vivo imaging of siRNA delivery and silencing in tumors. Nat Med 13, 372–377 (2007). https://doi.org/10.1038/nm1486
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DOI: https://doi.org/10.1038/nm1486
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