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In vivo imaging of siRNA delivery and silencing in tumors

Nature Medicine volume 13, pages 372–377 (2007)Cite this article

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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Figure 1: Synthesis of MN-NIRF-siGFP.
Figure 2: In vitro testing of MN-NIRF-siGFP cell uptake and silencing efficiency in stably transfected 9L-GFP gliosarcoma cells.
Figure 3: In vivo imaging of MN-NIRF-siGFP delivery to tumors.
Figure 4: In vivo imaging of MN-NIRF-siGFP silencing in tumors.
Figure 5: Application of MN-NIRF-siSurvivin in a therapeutic tumor model.

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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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Author notes

  1. Zdravka Medarova and Wellington Pham: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Radiology, Massachussetts General Hospital/Massachusetts Institute of Technology/Harvard Medical School Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital/Harvard Medical School, Room 2301, Building 149, 13th Street, Charlestown, Boston, 02129, Massachusetts, USA

    Zdravka Medarova, Wellington Pham, Christian Farrar & Anna Moore

  2. Beth Israel Deaconess Medical Center, Harvard Medical School, Room 1034, 77 Louis Pasteur Avenue, Boston, 02215, Massachusetts, USA

    Victoria Petkova

Authors
  1. Zdravka Medarova
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  2. Wellington Pham
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  3. Christian Farrar
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  4. Victoria Petkova
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  5. Anna Moore
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Contributions

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.

Corresponding author

Correspondence to Anna Moore.

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Competing interests

The authors declare no competing financial interests.

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)

Supplementary Methods (PDF 67 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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