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Microfluidic Assembly of siRNA-Loaded Micelleplexes for Tumor Targeting in an Orthotopic Model of Ovarian Cancer

  • Daniel P. Feldmann
  • Steven Jones
  • Kirk Douglas
  • Anthony F. Shields
  • Olivia M. MerkelEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1974)

Abstract

The use of cationic polymers to interact with negatively charged siRNA via charge complexation to form polyelectrolyte complexes has been widely studied ever since the 1998 report on RNA interference. These polyelectrolyte complex formulations aim to overcome the many pitfalls associated with the use of RNA interference as a potential cancer therapy. The triblock copolymer polyethylenimine-polycaprolactone-polyethylene glycol (PEI-PCL-PEG) contains the cation PEI and has been shown to be an efficient carrier capable of complexing with nucleic acids for gene delivery. This copolymer system also allows for targeting moieties to be linked to the micelleplex, thereby exploiting overexpressed receptors (such as the folate receptor) located within tumors. Additionally, we demonstrated recently that microfluidic mixing of PEI-PCL-PEG nanoparticles allows for the rapid, scaled-up production of micelleplexes while maintaining small and uniform particle distributions. The preparation of small and reproducible particles is imperative for clinical translation of nanomedicine and for tumor targeting via systemic administration. Furthermore, to enable tracing of its deposition in vivo after its administration, micelleplexes can be radiolabeled. To assess tumor targeting over time, the noninvasive imaging technique single-photon emission computed tomography (SPECT) offers the ability to examine the same subject at multiple time points and generate biodistribution profiles. Since the biodistribution and tumor targeting of the therapeutic load of micelleplexes is of foremost interest, we recently described an approach to modify siRNA with a DTPA (diethylenetriaminepentaacetic acid) chelator. Herein, we explain the details of encapsulating indium-labeled siRNA via microfluidic mixing in PEI-PCL-PEG nanoparticles with a folic acid targeting ligand for assessment of their in vivo tumor targeting in an orthotopic ovarian cancer model.

Keywords

siRNA delivery SPECT imaging Indium labeling Microfluidic mixing Triblock copolymer Folate receptor targeting 

Notes

Acknowledgments

Research reported in this publication was supported by the Wayne State University Nano Incubator grant to Olivia Merkel, the Ruth L. Kirschstein National Research Award T32-CA009531 fellowship to Steven Jones, and the Wayne State School of Medicine GRA support of Daniel Feldmann.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Daniel P. Feldmann
    • 1
  • Steven Jones
    • 1
  • Kirk Douglas
    • 2
  • Anthony F. Shields
    • 2
  • Olivia M. Merkel
    • 1
    • 3
    • 4
    Email author
  1. 1.Department of OncologyWayne State University School of MedicineDetroitUSA
  2. 2.Department of Oncology, Karmanos Cancer InstituteWayne State UniversityDetroitUSA
  3. 3.Department of Pharmaceutical SciencesWayne State UniversityDetroitUSA
  4. 4.Department of Pharmacy, Pharmaceutical Technology and BiopharmacyLudwig-Maximilians-Universität MünchenMunichGermany

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